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Sample records for solid-state nmr probe

  1. Probing Quadrupolar Nuclei by Solid-State NMR Spectroscopy: Recent Advances

    SciTech Connect

    Fernandez, Christian; Pruski, Marek

    2011-06-08

    Solid-state nuclear magnetic resonance (NMR) of quadrupolar nuclei has recently undergone remarkable development of capabilities for obtaining structural and dynamic information at the molecular level. This review summarizes the key achievements attained during the last couple of decades in solid-state NMR of both integer spin and half-integer spin quadrupolar nuclei. We provide a concise description of the first- and second-order quadrupolar interactions, and their effect on the static and magic angle spinning (MAS) spectra. Methods are explained for efficient excitation of single- and multiple-quantum coherences, and acquisition of spectra under low- and high-resolution conditions. Most of all, we present a coherent, comparative description of the high-resolution methods for half-integer quadrupolar nuclei, including double rotation (DOR), dynamic angle spinning (DAS), multiple-quantum magic angle spinning (MQMAS), and satellite transition magic angle spinning (STMAS). Also highlighted are methods for processing and analysis of the spectra. Finally, we review methods for probing the heteronuclear and homonuclear correlations between the quadrupolar nuclei and their quadrupolar or spin-1/2 neighbors.

  2. The ABC exporter MsbA probed by solid state NMR – challenges and opportunities.

    PubMed

    Kaur, Hundeep; Lakatos, Andrea; Spadaccini, Roberta; Vogel, Ramona; Hoffmann, Christian; Becker-Baldus, Johanna; Ouari, Olivier; Tordo, Paul; Mchaourab, Hassane; Glaubitz, Clemens

    2015-09-01

    ATP binding cassette (ABC) transporters form a superfamily of integral membrane proteins involved in translocation of substrates across the membrane driven by ATP hydrolysis. Despite available crystal structures and extensive biochemical data, many open questions regarding their transport mechanisms remain. Therefore, there is a need to explore spectroscopic techniques such as solid state NMR in order to bridge the gap between structural and mechanistic data. In this study, we investigate the feasibility of using Escherichia coli MsbA as a model ABC transporter for solid state NMR studies. We show that optimised solubilisation and reconstitution procedures enable preparing stable and homogenous protein samples. Depending on the duration of solubilisation, MsbA can be obtained in either an apo- or in a native lipid A bound form. Building onto these optimisations, the first promising MAS-NMR spectra with narrow lines have been recorded. However, further sensitivity improvements are required so that complex NMR experiments can be recorded within a reasonable amount of time. We therefore demonstrate the usability of paramagnetic doping for rapid data acquisition and explore dynamic nuclear polarisation as a method for general signal enhancement. Our results demonstrate that solid state NMR provides an opportunity to address important biological questions related to complex mechanisms of ABC transporters. PMID:25849794

  3. Probing the nanostructure of bioerodible polyanhydrides with solid-state NMR

    NASA Astrophysics Data System (ADS)

    Kipper, Matt; Hou, Sheng-Shu; Schmidt-Rohr, Klaus; Narasimhan, Balaji

    2003-03-01

    Polyanhydrides are a class of bioerodible polymers with unique properties that make them ideal for drug delivery applications. One of the most interesting properties of these materials is the microphase separation that occurs in copolymers of certain compositions. Microphase separation of copolymers, when coupled with the disparity in degradation rates between two constituent monomers, results in interesting degradation and drug release phenomena that can be exploited to design controlled release devices with tailored release profiles. Solid-state NMR spectroscopy is used to identify the crystalline domains of copolymers of sebacic Acid (SA) and 1,6-bis(p-carboxyphenoxy)hexane (CPH). Proton spin diffusion experiments with carbon detection are used to discern length scales of phase separation in the amorphous regions of the polymer. The data indicate that when the copolymer composition exceeds 75constituent co-monomers, nanoscale domains of the minority component are present, and a weakly segregated block-like nanostructure is obtained. As the copolymer composition nears 50These observations are consistent with solution state NMR data that measure the number-average sequence of the polymer chains.

  4. Solid-state NMR imaging system

    SciTech Connect

    Gopalsami, N.; Dieckman, S.L.; Ellingson, W.A.

    1990-01-01

    An accessory for use with a solid-state NMR spectrometer includes a special imaging probe with linear, high-field strength gradient fields and high-power broadband RF coils using a back projection method for data acquisition and image reconstruction, and a real-time pulse programmer adaptable for use by a conventional computer for complex high speed pulse sequences.

  5. Solid-state NMR imaging system

    DOEpatents

    Gopalsami, Nachappa; Dieckman, Stephen L.; Ellingson, William A.

    1992-01-01

    An apparatus for use with a solid-state NMR spectrometer includes a special imaging probe with linear, high-field strength gradient fields and high-power broadband RF coils using a back projection method for data acquisition and image reconstruction, and a real-time pulse programmer adaptable for use by a conventional computer for complex high speed pulse sequences.

  6. Probing membrane protein structure using water polarization transfer solid-state NMR

    NASA Astrophysics Data System (ADS)

    Williams, Jonathan K.; Hong, Mei

    2014-10-01

    Water plays an essential role in the structure and function of proteins, lipid membranes and other biological macromolecules. Solid-state NMR heteronuclear-detected 1H polarization transfer from water to biomolecules is a versatile approach for studying water-protein, water-membrane, and water-carbohydrate interactions in biology. We review radiofrequency pulse sequences for measuring water polarization transfer to biomolecules, the mechanisms of polarization transfer, and the application of this method to various biological systems. Three polarization transfer mechanisms, chemical exchange, spin diffusion and NOE, manifest themselves at different temperatures, magic-angle-spinning frequencies, and pulse irradiations. Chemical exchange is ubiquitous in all systems examined so far, and spin diffusion plays the key role in polarization transfer within the macromolecule. Tightly bound water molecules with long residence times are rare in proteins at ambient temperature. The water polarization-transfer technique has been used to study the hydration of microcrystalline proteins, lipid membranes, and plant cell wall polysaccharides, and to derive atomic-resolution details of the kinetics and mechanism of ion conduction in channels and pumps. Using this approach, we have measured the water polarization transfer to the transmembrane domain of the influenza M2 protein to obtain information on the structure of this tetrameric proton channel. At short mixing times, the polarization transfer rates are site-specific and depend on the pH, labile protons, sidechain conformation, as well as the radial position of the residues in this four-helix bundle. Despite the multiple dependences, the initial transfer rates reflect the periodic nature of the residue positions from the water-filled pore, thus this technique provides a way of gleaning secondary structure information, helix tilt angle, and the oligomeric structure of membrane proteins.

  7. Probing Membrane Protein Structure Using Water Polarization Transfer Solid-State NMR

    PubMed Central

    Williams, Jonathan K.; Hong, Mei

    2014-01-01

    Water plays an essential role in the structure and function of proteins, lipid membranes and other biological macromolecules. Solid-state NMR heteronuclear-detected 1H polarization transfer from water to biomolecules is a versatile approach for studying water-protein, water-membrane, and water-carbohydrate interactions in biology. We review radiofrequency pulse sequences for measuring water polarization transfer to biomolecules, the mechanisms of polarization transfer, and the application of this method to various biological systems. Three polarization transfer mechanisms, chemical exchange, spin diffusion and NOE, manifest themselves at different temperatures, magic-angle-spinning frequencies, and pulse irradiations. Chemical exchange is ubiquitous in all systems examined so far, and spin diffusion plays the key role in polarization transfer within the macromolecule. Tightly bound water molecules with long residence times are rare in proteins at ambient temperature. The water polarization-transfer technique has been used to study the hydration of microcrystalline proteins, lipid membranes, and plant cell wall polysaccharides, and to derive atomic-resolution details of the kinetics and mechanism of ion conduction in channels and pumps. Using this approach, we have measured the water polarization transfer to the transmembrane peptide of the influenza M2 protein to obtain information on the structure of this tetrameric proton channel. At short mixing times, the polarization transfer rates are site-specific and depend on the pH, labile protons, sidechain conformation, as well as the radial position of the residues in this four-helix bundle. Despite the multiple dependences, the initial transfer rates reflect the periodic nature of the residue positions from the water-filled pore, thus this technique provides a way of gleaning secondary structure information, helix tilt angle, and the oligomeric structure of membrane proteins. PMID:25228502

  8. Dynamics of Hydrophobic Core Phenylalanine Residues Probed by Solid-State Deuteron NMR

    SciTech Connect

    Vugmeyster, Liliya; Ostrovsky, Dmitry; Villafranca, Toni; Sharp, Janelle; Xu, Wei; Lipton, Andrew S.; Hoatson, Gina; Vold, Robert L.

    2015-11-03

    We conducted a detailed investigation of the dynamics of two phenylalanine side chains in the hydrophobic core of the villin headpiece subdomain protein (HP36) in the hydrated powder state over the 298–80 K temperature range. We utilized static deuteron NMR measurements of longitudinal relaxation and line shapes supplemented with computational modeling. The temperature dependence of the relaxation times reveals the presence of two main mechanisms that can be attributed to the ring-flips, dominating at high temperatures, and small-angle fluctuations, dominating at low temperatures. The relaxation is non- exponential at all temperatures with the extent of non-exponentiality increasing from higher to lower temperatures. This behavior suggests a distribution of conformers with unique values of activation energies. The central values of the activation energies for the ring-flipping motions are among the smallest reported for aromatic residues in peptides and proteins and point to a very mobile hydrophobic core. The analysis of the widths of the distributions, in combination with the earlier results on the dynamics of flanking methyl groups (Vugmeyster et al., J. Phys. Chem. 2013, 117, 6129–6137), suggests that the hydrophobic core undergoes concerted fluctuations. There is a pronounced effect of dehydration on the ring-flipping motions, which shifts the distribution toward more rigid conformers. The cross-over temperature between the regions of dominance of the small-angle fluctuations and ring-flips shifts from 195 K in the hydrated protein to 278 K in the dry one. This result points to the role of solvent in the onset of the concerted fluctuations of the core and highlights aromatic residues as markers of the protein dynamical transitions.

  9. Solid-state NMR and Membrane Proteins

    PubMed Central

    Opella, Stanley J.

    2015-01-01

    The native environment for a membrane protein is a phospholipid bilayer. Because the protein is immobilized on NMR timescales by the interactions within a bilayer membrane, solid-state NMR methods are essential to obtain high-resolution spectra. Approaches have been developed for both unoriented and oriented samples, however, they all rest on the foundation of the most fundamental aspects solid-state NMR, and the chemical shift and homo- and hetero-nuclear dipole-dipole interactions. Solid-state NMR has advanced sufficiently to enable the structures of membrane proteins to be determined under near-native conditions in phospholipid bilayers. PMID:25681966

  10. REDOR solid-state NMR as a probe of the membrane locations of membrane-associated peptides and proteins

    NASA Astrophysics Data System (ADS)

    Jia, Lihui; Liang, Shuang; Sackett, Kelly; Xie, Li; Ghosh, Ujjayini; Weliky, David P.

    2015-04-01

    Rotational-echo double-resonance (REDOR) solid-state NMR is applied to probe the membrane locations of specific residues of membrane proteins. Couplings are measured between protein 13CO nuclei and membrane lipid or cholesterol 2H and 31P nuclei. Specific 13CO labeling is used to enable unambiguous assignment and 2H labeling covers a small region of the lipid or cholesterol molecule. The 13CO-31P and 13CO-2H REDOR respectively probe proximity to the membrane headgroup region and proximity to specific insertion depths within the membrane hydrocarbon core. One strength of the REDOR approach is use of chemically-native proteins and membrane components. The conventional REDOR pulse sequence with 100 kHz 2H π pulses is robust with respect to the 2H quadrupolar anisotropy. The 2H T1's are comparable to the longer dephasing times (τ's) and this leads to exponential rather than sigmoidal REDOR buildups. The 13CO-2H buildups are well-fitted to A × (1 - e-γτ) where A and γ are fitting parameters that are correlated as the fraction of molecules (A) with effective 13CO-2H coupling d = 3γ/2. The REDOR approach is applied to probe the membrane locations of the "fusion peptide" regions of the HIV gp41 and influenza virus hemagglutinin proteins which both catalyze joining of the viral and host cell membranes during initial infection of the cell. The HIV fusion peptide forms an intermolecular antiparallel β sheet and the REDOR data support major deeply-inserted and minor shallowly-inserted molecular populations. A significant fraction of the influenza fusion peptide molecules form a tight hairpin with antiparallel N- and C-α helices and the REDOR data support a single peptide population with a deeply-inserted N-helix. The shared feature of deep insertion of the β and α fusion peptide structures may be relevant for fusion catalysis via the resultant local perturbation of the membrane bilayer. Future applications of the REDOR approach may include samples that contain cell membrane extracts and use of lower temperatures and dynamic nuclear polarization to reduce data acquisition times.

  11. Probing hydrogen bonding in cocrystals and amorphous dispersions using (14)N-(1)H HMQC solid-state NMR.

    PubMed

    Tatton, Andrew S; Pham, Tran N; Vogt, Frederick G; Iuga, Dinu; Edwards, Andrew J; Brown, Steven P

    2013-03-01

    Cocrystals and amorphous solid dispersions have generated interest in the pharmaceutical industry as an alternative to more established solid delivery forms. The identification of intermolecular hydrogen bonding interactions in a nicotinamide palmitic acid cocrystal and a 50% w/w acetaminophen-polyvinylpyrrolidone solid dispersion are reported using advanced solid-state magic-angle spinning (MAS) NMR methods. The application of a novel (14)N-(1)H HMQC experiment, where coherence transfer is achieved via through-space couplings, is shown to identify specific hydrogen bonding motifs. Additionally, (1)H isotropic chemical shifts and (14)N electric field gradient (EFG) parameters, both accessible from (14)N-(1)H HMQC experiments, are shown to be sensitive to changes in hydrogen bonding geometry. Numerous indicators of molecular association are accessible from this experiment, including NH cross-peaks occurring from intermolecular hydrogen bonds and changes in proton chemical shifts or electric field gradient parameters. First-principles calculations using the GIPAW approach that yield accurate estimates of isotropic chemical shifts, and EFG parameters were used to assist in assignment. It is envisaged that (14)N-(1)H HMQC solid state NMR experiments could become a valuable screening technique of solid delivery forms in the pharmaceutical industry. PMID:23301555

  12. A Modified Alderman-Grant Coil makes possible an efficient cross-coil probe for high field solid-state NMR of lossy biological samples

    NASA Astrophysics Data System (ADS)

    Grant, Christopher V.; Yang, Yuan; Glibowicka, Mira; Wu, Chin H.; Park, Sang Ho; Deber, Charles M.; Opella, Stanley J.

    2009-11-01

    The design, construction, and performance of a cross-coil double-resonance probe for solid-state NMR experiments on lossy biological samples at high magnetic fields are described. The outer coil is a Modified Alderman-Grant Coil (MAGC) tuned to the 1H frequency. The inner coil consists of a multi-turn solenoid coil that produces a B 1 field orthogonal to that of the outer coil. This results in a compact nested cross-coil pair with the inner solenoid coil tuned to the low frequency detection channel. This design has several advantages over multiple-tuned solenoid coil probes, since RF heating from the 1H channel is substantially reduced, it can be tuned for samples with a wide range of dielectric constants, and the simplified circuit design and high inductance inner coil provides excellent sensitivity. The utility of this probe is demonstrated on two electrically lossy samples of membrane proteins in phospholipid bilayers (bicelles) that are particularly difficult for conventional NMR probes. The 72-residue polypeptide embedding the transmembrane helices 3 and 4 of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) (residues 194-241) requires a high salt concentration in order to be successfully reconstituted in phospholipid bicelles. A second application is to paramagnetic relaxation enhancement applied to the membrane-bound form of Pf1 coat protein in phospholipid bicelles where the resistance to sample heating enables high duty cycle solid-state NMR experiments to be performed.

  13. Scalar operators in solid-state NMR

    SciTech Connect

    Sun, Boqin

    1991-11-01

    Selectivity and resolution of solid-state NMR spectra are determined by dispersion of local magnetic fields originating from relaxation effects and orientation-dependent resonant frequencies of spin nuclei. Theoretically, the orientation-dependent resonant frequencies can be represented by a set of irreducible tensors. Among these tensors, only zero rank tensors (scalar operators) are capable of providing high resolution NMR spectra. This thesis presents a series of new developments in high resolution solid-state NMR concerning the reconstruction of various scalar operators motion in solid C{sub 60} is analyzed.

  14. Probing physical and chemical changes in cortical bone due to osteoporosis and type 2 diabetes by solid-state NMR

    NASA Astrophysics Data System (ADS)

    Zhou, Donghua; Taylor, Amanda; Rendina, Beth; Smith, Brenda; Department of Physics Collaboration; Department of Nutritional Sciences Collaboration

    2013-03-01

    Approximately 1.5 million fractures occur each year in the U.S. due to osteoporosis, which is characterized by decreased bone mineral density and deterioration of bone micro-architecture. On the other hand, type 2 diabetes also significantly increases fracture risks, despite having a normal or even higher bone mineral density. Solid-state NMR has been applied to bone tissues from normal and disease-inflicted mouse models to study structural and chemical dynamics as the disease progresses. Proton relaxation experiments were performed to measure water populations in the bone matrix and pores. Collagen-bound water has strong influence on bone resilience, while water content in the pores reveals amount and size of pores from micro- to millimeter range. Other biochemical and atomic-scale structural alterations in the mineral and organic phases and their interface were investigated by proton, phosphorus, and carbon NMR spectroscopy. Experiments were designed to individually detect different types of phosphorus environments: near the mineral surface, similar to hydroxyapatite, and deficient of hydrogens due to substitution of the hydroxyl group by other ions. A new method was also developed for accurate quantification of each phosphorus species. The authors appreciate financial support for this project from the College of Human Sciences and the College of Arts and Sciences, Oklahoma State University.

  15. Solid-State NMR of PEGylated Proteins.

    PubMed

    Ravera, Enrico; Ciambellotti, Silvia; Cerofolini, Linda; Martelli, Tommaso; Kozyreva, Tatiana; Bernacchioni, Caterina; Giuntini, Stefano; Fragai, Marco; Turano, Paola; Luchinat, Claudio

    2016-02-01

    PEGylated proteins are widely used in biomedicine but, in spite of their importance, no atomic-level information is available since they are generally resistant to structural characterization approaches. PEGylated proteins are shown here to yield highly resolved solid-state NMR spectra, which allows assessment of the structural integrity of proteins when PEGylated for therapeutic or diagnostic use. PMID:26756539

  16. Solid-State NMR for Bacterial Biofilms

    PubMed Central

    Reichhardt, Courtney; Cegelski, Lynette

    2014-01-01

    Bacteria associate with surfaces and one another by elaborating an extracellular matrix to encapsulate cells, creating communities termed biofilms. Biofilms are beneficial in some ecological niches, but also contribute to the pathogenesis of serious and chronic infectious diseases. New approaches and quantitative measurements are needed to define the composition and architecture of bacterial biofilms to help drive the development of strategies to interfere with biofilm assembly. Solid-state NMR is uniquely suited to the examination of insoluble and complex macromolecular and whole-cell systems. This article highlights three examples that implement solid-state NMR to deliver insights into bacterial biofilm composition and changes in cell-wall composition as cells transition to the biofilm lifestyle. Most recently, solid-state NMR measurements provided a total accounting of the protein and polysaccharide components in the extracellular matrix of an E. coli biofilm and transform our qualitative descriptions of matrix composition into chemical parameters that permit quantitative comparisons among samples. We present additional data for whole biofilm samples (cells plus the extracellular matrix) that complement matrix-only analyses. The study of bacterial biofilms by solid-state NMR is an exciting avenue ripe with many opportunities and we close the article by articulating some outstanding questions and future directions in this area. PMID:24976646

  17. Conformational dynamics of a seven transmembrane helical protein Anabaena Sensory Rhodopsin probed by solid-state NMR.

    PubMed

    Good, Daryl B; Wang, Shenlin; Ward, Meaghan E; Struppe, Jochem; Brown, Leonid S; Lewandowski, Józef R; Ladizhansky, Vladimir

    2014-02-19

    The ability to detect and characterize molecular motions represents one of the unique strengths of nuclear magnetic resonance (NMR) spectroscopy. In this study, we report solid-state NMR site-specific measurements of the dipolar order parameters and (15)N rotating frame spin-lattice (R1ρ) relaxation rates in a seven transmembrane helical protein Anabaena Sensory Rhodopsin reconstituted in lipids. The magnitudes of the observed order parameters indicate that both the well-defined transmembrane regions and the less structured intramembrane loops undergo restricted submicrosecond time scale motions. In contrast, the R1ρ rates, which were measured under fast magic angle spinning conditions, vary by an order of magnitude between the TM and exposed regions and suggest the presence of intermediate time scale motions. Using a simple model, which assumes a single exponential autocorrelation function, we estimated the time scales of dominant stochastic motions to be on the order of low tens of nanoseconds for most residues within the TM helices and tens to hundreds of nanoseconds for the extracellular B-C and F-G loops. These relatively slow time scales could be attributed to collective anisotropic motions. We used the 3D Gaussian axial fluctuations model to estimate amplitudes, directions, and time scales of overall motions for helices and the extracellular B-C and F-G loops. Within this model, the TM helices A,B,C,D,E,F undergo rigid body motions on a time scale of tens of nanoseconds, while the time scale for the seventh helix G approaches 100 ns. Similar time scales of roughly 100-200 ns are estimated for the B-C and F-G loops. PMID:24467417

  18. Probing the Transmembrane Structure and Dynamics of Microsomal NADPH-cytochrome P450 oxidoreductase by Solid-State NMR

    PubMed Central

    Huang, Rui; Yamamoto, Kazutoshi; Zhang, Meng; Popovych, Nataliya; Hung, Ivan; Im, Sang-Choul; Gan, Zhehong; Waskell, Lucy; Ramamoorthy, Ayyalusamy

    2014-01-01

    NADPH-cytochrome P450 oxidoreductase (CYPOR) is an essential redox partner of the cytochrome P450 (cyt P450) superfamily of metabolic enzymes. In the endoplasmic reticulum of liver cells, such enzymes metabolize ∼75% of the pharmaceuticals in use today. It is known that the transmembrane domain of CYPOR plays a crucial role in aiding the formation of a complex between CYPOR and cyt P450. Here we present the transmembrane structure, topology, and dynamics of the FMN binding domain of CYPOR in a native membrane-like environment. Our solid-state NMR results reveal that the N-terminal transmembrane domain of CYPOR adopts an α-helical conformation in the lipid membrane environment. Most notably, we also show that the transmembrane helix is tilted ∼13° from the lipid bilayer normal, and exhibits motions on a submillisecond timescale including rotational diffusion of the whole helix and fluctuation of the helical director axis. The approaches and the information reported in this study would enable further investigations on the structure and dynamics of the full-length NADPH-cytochrome P450 oxidoreductase and its interaction with other membrane proteins in a membrane environment. PMID:24853741

  19. High-Resolution Solid-State NMR

    NASA Astrophysics Data System (ADS)

    Massiot, Dominique

    High-resolution solid state NMR and its application to the characterization of solid state inorganic, organic, or hybrid materials, is undergoing rapid developments on both methodological and hardware points of view. Starting from a general presentation, we describe different methods that give access to the high resolution spectra; we then give examples of new methods that enable more detailed structural description through the selective reintroduction of anisotropic interactions (quadrupolar, chemical shift anisotropy, dipolar and scalar couplings). A special emphasis is given to the recent improvements obtained at high, very-high and ultra-high magnetic fields.

  20. Solid-state NMR studies of supercapacitors.

    PubMed

    Griffin, John M; Forse, Alexander C; Grey, Clare P

    2016-01-01

    Electrochemical double-layer capacitors, or 'supercapacitors' are attracting increasing attention as high-power energy storage devices for a wide range of technological applications. These devices store charge through electrostatic interactions between liquid electrolyte ions and the surfaces of porous carbon electrodes. However, many aspects of the fundamental mechanism of supercapacitance are still not well understood, and there is a lack of experimental techniques which are capable of studying working devices. Recently, solid-state NMR has emerged as a powerful tool for studying the local environments and behaviour of electrolyte ions in supercapacitor electrodes. In this Trends article, we review these recent developments and applications. We first discuss the basic principles underlying the mechanism of supercapacitance, as well as the key NMR observables that are relevant to the study of supercapacitor electrodes. We then review some practical aspects of the study of working devices using ex situ and in situ methodologies and explain the key advances that these techniques have allowed on the study of supercapacitor charging mechanisms. NMR experiments have revealed that the pores of the carbon electrodes contain a significant number of electrolyte ions in the absence of any charging potential. This has important implications for the molecular mechanisms of supercapacitance, as charge can be stored by different ion adsorption/desorption processes. Crucially, we show how in situ NMR experiments can be used to quantitatively study and characterise the charging mechanism, with the experiments providing the most detailed picture of charge storage to date, offering the opportunity to design enhanced devices. Finally, an outlook for future directions for solid-state NMR in supercapacitor research is offered. PMID:26974032

  1. Lithium Polymer Electrolytes and Solid State NMR

    NASA Technical Reports Server (NTRS)

    Berkeley, Emily R.

    2004-01-01

    Research is being done at the Glenn Research Center (GRC) developing new kinds of batteries that do not depend on a solution. Currently, batteries use liquid electrolytes containing lithium. Problems with the liquid electrolyte are (1) solvents used can leak out of the battery, so larger, more restrictive, packages have to be made, inhibiting the diversity of application and decreasing the power density; (2) the liquid is incompatible with the lithium metal anode, so alternative, less efficient, anodes are required. The Materials Department at GRC has been working to synthesize polymer electrolytes that can replace the liquid electrolytes. The advantages are that polymer electrolytes do not have the potential to leak so they can be used for a variety of tasks, small or large, including in the space rover or in space suits. The polymers generated by Dr. Mary Ann Meador's group are in the form of rod -coil structures. The rod aspect gives the polymer structural integrity, while the coil makes it flexible. Lithium ions are used in these polymers because of their high mobility. The coils have repeating units of oxygen which stabilize the positive lithium by donating electron density. This aids in the movement of the lithium within the polymer, which contributes to higher conductivity. In addition to conductivity testing, these polymers are characterized using DSC, TGA, FTIR, and solid state NMR. Solid state NMR is used in classifying materials that are not soluble in solvents, such as polymers. The NMR spins the sample at a magic angle (54.7') allowing the significant peaks to emerge. Although solid state NMR is a helpful technique in determining bonding, the process of preparing the sample and tuning it properly are intricate jobs that require patience; especially since each run takes about six hours. The NMR allows for the advancement of polymer synthesis by showing if the expected results were achieved. Using the NMR, in addition to looking at polymers, allows for participation on a variety of other projects, including aero-gels and carbon graphite mat en als. The goals of the polymer electrolyte research are to improve the physical properties of the polymers. This includes improving conductivity, durability, and expanding the temperature range over which it is effective. Currently, good conductivity is only present at high temperatures. My goals are to experiment with different arrangements of rods and coils to achieve these desirable properties. Some of my experiments include changing the number of repeat units in the polymer, the size of the diamines, and the types of coil. Analysis of these new polymers indicates improvement in some properties, such as lower glass transition temperature; however, they are not as flexible as desired. With further research we hope to produce polymers that encompass all of these properties to a high degree.

  2. Quasi equilibria in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Sakellariou, Dimitris; Hodgkinson, Paul; Emsley, Lyndon

    1998-08-01

    We develop the connection between theoretical studies of so-called quasi-equilibrium states in solid-state NMR for isolated spin-systems with the experimental observability of such states. The observability of these states is expected to be dependent on the relative broadenings due to coherent effects (such as dipolar couplings with the lattice) and incoherent effects, i.e. relaxation. Hence, we expect to see evidence of quasi-equilibria in cross polarization experiments, where the relevant relaxation time constant is relatively long. For classic spectral spin-diffusion experiments, however, quasi equilibria are unlikely to be observed. We also generalise the discussion to rotating solids, which is appropriate to magic angle spinning experiments.

  3. CONFORMATIONAL FLEXIBILITY AND STRAND ARRANGEMENTS OF THE MEMBRANE-ASSOCIATED HIV FUSION PEPTIDE TRIMER PROBED BY SOLID-STATE NMR SPECTROSCOPY†

    PubMed Central

    Zheng, Zhaoxiong; Yang, Rong; Bodner, Michele L.; Weliky, David P.

    2008-01-01

    The human immunodeficiency virus (HIV) fusion peptide (HFP) is the N-terminal apolar region of the HIV gp41 fusion protein and interacts with target cell membranes and promotes membrane fusion. The free peptide catalyzes vesicle fusion at least to the lipid mixing stage and serves as a useful model fusion system. For gp41 constructs which lack the HFP, high-resolution structures show trimeric protein and suggest that at least three HFPs interact with the membrane with their C-termini in close proximity. In addition, previous studies have demonstrated that HFPs which are cross-linked at their C-termini to form trimers (HFPtr) catalyze fusion at a rate which is 15−40 times greater than non-cross-linked HFP. In the present study, the structure of membrane-associated HFPtr was probed with solid-state nuclear magnetic resonance (NMR) methods. Chemical shift and intramolecular 13CO-15N distance measurements show that the conformation of the Leu-7 to Phe-11 region of HFPtr has predominant helical conformation in membranes without cholesterol and β strand conformation in membranes containing ∼30 mol% cholesterol. Interstrand 13CO-13CO and 13CO-15N distance measurements were not consistent with an in-register parallel strand arrangement but were consistent with either: (1) parallel arrangement with adjacent strands two-residues out-of-register; or (2) antiparallel arrangement with adjacent strand crossing between Phe-8 and Leu-9. Arrangement (1) could support the rapid fusion rate of HFPtr because of placement of the apolar N-terminal regions of all strands on the same side of the oligomer while arrangement (2) could support the assembly of multiple fusion protein trimers. PMID:17059213

  4. Probing the structural disorder of basalts and slab-driven andesite melts: Insights from high-resolution solid-state NMR study

    NASA Astrophysics Data System (ADS)

    Park, S.; Lee, S.

    2012-12-01

    Whereas the structure of multi-component silicate melts has strong implication for the properties of natural silicate melts and relevant magmatic processes in Earth's mantle and crust, little is known about their atomic structures due to lack of suitable experimental probes of multi-component amorphous oxides. Although most of the progress in melt structure has been made for relatively simple binary and ternary silicate glasses, recent advances in high-resolution solid-state NMR (nuclear magnetic resonance) unveil previously unknown structural details of multi-component silicate melts (Lee, S. K. and Sung, S., Chem. Geol., 2008, 256, 326; Lee et al., P. Natl. Acad. Sci. USA., 2011, 108, 6847; Park and Lee, Geochim. Cosmochim. Acta, 2012, 80, 125). In this study we report experimental results on the effects of composition. atomic structure of CaO-MgO-Al_{2} O_{3} -SiO_{2} (CMAS) glasses in diopside (CaMgSi_{2}O_{6}) and Ca-tschermakite (CaAl_{2}SiO_{6}) join and glass in the diopside-anorthite eutectic composition (Di_{64}An_{36})—model systems for basaltic melts—using solid-state NMR. We also report the first high-resolution experimental results on the atomic structure of CaO-MgO-Na_{2}O-Al_{2}O_{3}-SiO_{2} (CMNAS) glasses in diopside and jadeite (NaAlSi_{2}O_{6}) join, and glass in the natural phonolite composition (CaO: MgO: Na_{2}O: K_{2}O: Al_{2}O_{3}: SiO_{2}= 1.4: 8.0: 9.0: 3.8: 13: 64 mol%), a model system for slab driven andesite melts. The Al-27 3QMAS (triple quantum magic angle spinning) NMR spectra of CMAS glasses in diopside-Ca-tschermakite join show predominant ^{[4]}Al and a non-negligible fraction of ^{[5]}Al. Approximately 3.3% of ^{[5]}Al is observed for Di_{64}An_{36} glass. The Al-27 3QMAS NMR spectra of CMNAS glasses in diopside and jadeite join show mostly ^{[4]}Al and a non-negligible fraction of ^{[5]}Al (X_{Diopside}=0.75, the mole fraction of diopside content). While the C_{q} (quadrupolar coupling constant) of ^{[4]}Al for glasses in diopside-Ca-tschermakite join decreases, that of ^{[4]}Al in diopside-jadeite increases with increasing X_{Diopside}. The Al-27 3QMAS NMR spectrum of phonolite glass shows only ^{[4]}Al species. The C_{q} of ^{[4]}Al ( 4.6 MHz) in phonolite glass is smaller than that of Di_{64}An_{36} eutectic glass ( 6.0 MHz) indicating larger deviation from perfect tetrahedral symmetry around ^{[4]}Al in basaltic melt than that of natural andesitic melt. The fraction of ^{[5]}Al, the degree of configurational disorder, increases with increasing X_{Ca+Mg} (the fraction of high field strength cations, X_{Ca+Mg}=[Ca+Mg]/[Ca+Mg+Na]) and thus the average cation field strength of non- framework cations (the average value of cation field strength normalized by mole fraction of each cations). The fraction of ^{[5]}Al is negligible from X_{Ca+Mg} =0 to X_{Ca+Mg} =0.08, and then it abruptly increases from 1.6 % for X_{Ca+Mg}=0.11 to 3.5% for X_{Ca+Mg}=0.13. This result indicates that there is a threshold composition (X_{Ca+Mg} 0.1) of silicate glasses and melts for increasing fraction of ^{[5]}Al. The abrupt changes of the fraction of ^{[5]}Al multi-component silicate glasses and melts can provide insight into drastic changes of macroscopic properties (entropy, viscosity, diffusivity, etc.) with varying composition of melt (i.e., basalt and andesite melts in this study).

  5. Calcium-43 chemical shift tensors as probes of calcium binding environments. Insight into the structure of the vaterite CaCO3 polymorph by 43Ca solid-state NMR spectroscopy.

    PubMed

    Bryce, David L; Bultz, Elijah B; Aebi, Dominic

    2008-07-23

    Natural-abundance (43)Ca solid-state NMR spectroscopy at 21.1 T and gauge-including projector-augmented-wave (GIPAW) DFT calculations are developed as tools to provide insight into calcium binding environments, with special emphasis on the calcium chemical shift (CS) tensor. The first complete analysis of a (43)Ca solid-state NMR spectrum, including the relative orientation of the CS and electric field gradient (EFG) tensors, is reported for calcite. GIPAW calculations of the (43)Ca CS and EFG tensors for a series of small molecules are shown to reproduce experimental trends; for example, the trend in available solid-state chemical shifts is reproduced with a correlation coefficient of 0.983. The results strongly suggest the utility of the calcium CS tensor as a novel probe of calcium binding environments in a range of calcium-containing materials. For example, for three polymorphs of CaCO3 the CS tensor span ranges from 8 to 70 ppm and the symmetry around calcium is manifested differently in the CS tensor as compared with the EFG tensor. The advantages of characterizing the CS tensor are particularly evident in very high magnetic fields where the effect of calcium CS anisotropy is augmented in hertz while the effect of second-order quadrupolar broadening is often obscured for (43)Ca because of its small quadrupole moment. Finally, as an application of the combined experimental-theoretical approach, the solid-state structure of the vaterite polymorph of calcium carbonate is probed and we conclude that the hexagonal P6(3)/mmc space group provides a better representation of the structure than does the orthorhombic Pbnm space group, thereby demonstrating the utility of (43)Ca solid-state NMR as a complementary tool to X-ray crystallographic methods. PMID:18576634

  6. Solid State NMR Investigations of Zeolite - Intercalate Structures

    SciTech Connect

    Fyfe, Colin A.; Diaz, Anix; Brouwer, Darren H.; Lee, Joseph; Schneider, Celine M.; Scheffler, Franziska A.; Darton, Richard J.

    2006-07-24

    We will describe two topics in which structural information on complexes of zeolites is obtained from solid state NMR. In the first, recent work on the determination of the complete three-dimensional structures of the complexes of zeolites with organic sorbates will be briefly reported. The method has been optimized and the presentation of the results systematized. In the second topic, we will describe how solid state NMR can be used in the reverse sense to probe for the existence and structures of “nanocrystals” whose dimensions are too small to give proper Bragg scattering and which have been proposed to be the synthesis route for the formation of zeolite ZSM-5. In this study, the spectral parameters of “probe” template molecules are used as being diagnostic of whether the local environment of the framework has been formed. These are independent of the “crystal” dimensions and this general approach may be applicable to other similar “nano” systems.

  7. Instrumentation for solid-state dynamic nuclear polarization with magic angle spinning NMR

    NASA Astrophysics Data System (ADS)

    Rosay, Melanie; Blank, Monica; Engelke, Frank

    2016-03-01

    Advances in dynamic nuclear polarization (DNP) instrumentation and methodology have been key factors in the recent growth of solid-state DNP NMR applications. We review the current state of the art of solid-state DNP NMR instrumentation primarily based on available commercial platforms. We start with a general system overview, including options for microwave sources and DNP NMR probes, and then focus on specific developments for DNP at 100 K with magic angle spinning (MAS). Gyrotron microwave sources, passive components to transmit microwaves, the DNP MAS probe, a cooling device for low-temperature MAS, and sample preparation procedures including radicals for DNP are considered.

  8. Instrumentation for solid-state dynamic nuclear polarization with magic angle spinning NMR.

    PubMed

    Rosay, Melanie; Blank, Monica; Engelke, Frank

    2016-03-01

    Advances in dynamic nuclear polarization (DNP) instrumentation and methodology have been key factors in the recent growth of solid-state DNP NMR applications. We review the current state of the art of solid-state DNP NMR instrumentation primarily based on available commercial platforms. We start with a general system overview, including options for microwave sources and DNP NMR probes, and then focus on specific developments for DNP at 100K with magic angle spinning (MAS). Gyrotron microwave sources, passive components to transmit microwaves, the DNP MAS probe, a cooling device for low-temperature MAS, and sample preparation procedures including radicals for DNP are considered. PMID:26920834

  9. Ultra-wideline solid-state NMR spectroscopy.

    PubMed

    Schurko, Robert W

    2013-09-17

    Although solid-state NMR (SSNMR) provides rich information about molecular structure and dynamics, the small spin population differences between pairs of spin states that give rise to NMR transitions make it an inherently insensitive spectroscopic technique in terms of signal acquisition. Scientists have continuously addressed this issue via improvements in NMR hardware and probes, increases in the strength of the magnetic field, and the development of innovative pulse sequences and acquisition methodologies. As a result, researchers can now study NMR-active nuclides previously thought to be unobservable or too unreceptive for routine examination via SSNMR. Several factors can make it extremely challenging to detect signal or acquire spectra using SSNMR: (i) low gyromagnetic ratios (i.e., low Larmor frequencies), (ii) low natural abundances or dilution of the nuclide of interest (e.g., metal nuclides in proteins or in organometallic catalysts supported on silica), (iii) inconvenient relaxation characteristics (e.g., very long longitudinal or very short transverse relaxation times), and/or (iv) extremely broad powder patterns arising from large anisotropic NMR interactions. Our research group has been particularly interested in efficient acquisition of broad NMR powder patterns for a variety of spin-1/2 and quadrupolar (spin > 1/2) nuclides. Traditionally, researchers have used the term "wideline" NMR to refer to experiments yielding broad (1)H and (2)H SSNMR spectra ranging from tens of kHz to ∼250 kHz in breadth. With modern FT NMR hardware, uniform excitation in these spectral ranges is relatively easy, allowing for the acquisition of high quality spectra. However, spectra that range in breadth from ca. 250 kHz to tens of MHz cannot be uniformly excited with conventional, high-power rectangular pulses. Rather, researchers must apply special methodologies to acquire such spectra, which have inherently low S/N because the signal intensity is spread across such large spectral breadths. We have suggested the term ultra-wideline NMR (UWNMR) spectroscopy to describe this set of methodologies. This Account describes recent developments in pulse sequences and strategies for the efficient acquisition of UWNMR spectra. After an introduction to anisotropically broadened NMR patterns, we give a brief history of methods used to acquire UWNMR spectra. We then discuss new acquisition methodologies, including the acquisition of CPMG echo trains and the application of pulses capable of broadband excitation and refocusing. Finally, we present several applications of UWNMR methods that use these broadband pulses. PMID:23745683

  10. Functional groups identified by solid state 13C NMR spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Animal manure is generally high in organic matter intensity so it is well suitable for 13C nuclear magnetic resonance (NMR) analysis. Solid-state 13C NMR techniques used in characterizing organic matter and its components include, but are not limited to, cross-polarization /magic angle spinning (CP...

  11. Probing the Transmembrane Structure and Topology of Microsomal Cytochrome-P450 by Solid-State NMR on Temperature-Resistant Bicelles

    PubMed Central

    Yamamoto, Kazutoshi; Gildenberg, Melissa; Ahuja, Shivani; Im, Sang-Choul; Pearcy, Paige; Waskell, Lucy; Ramamoorthy, Ayyalusamy

    2013-01-01

    Though the importance of high-resolution structure and dynamics of membrane proteins has been well recognized, optimizing sample conditions to retain the native-like folding and function of membrane proteins for Nuclear Magnetic Resonance (NMR) or X-ray measurements has been a major challenge. While bicelles have been shown to stabilize the function of membrane proteins and are increasingly utilized as model membranes, the loss of their magnetic-alignment at low temperatures makes them unsuitable to study heat-sensitive membrane proteins like cytochrome-P450 and protein-protein complexes. In this study, we report temperature resistant bicelles that can magnetically-align for a broad range of temperatures and demonstrate their advantages in the structural studies of full-length microsomal cytochrome-P450 and cytochrome-b5 by solid-state NMR spectroscopy. Our results reveal that the N-terminal region of rabbit cytochromeP4502B4, that is usually cleaved off to obtain crystal structures, is helical and has a transmembrane orientation with ~17° tilt from the lipid bilayer normal. PMID:23989972

  12. The development of solid-state NMR of membrane proteins

    PubMed Central

    Opella, Stanley J.

    2014-01-01

    Most biological functions are carried out in supramolecular assemblies. As a result of their slow reorientation in solution, these assemblies have been resistant to the widely employed solution NMR approaches. The development of solid-state NMR to first of all overcome the correlation time problem and then obtain informative high-resolution spectra of proteins in supramolecular assemblies, such as virus particles and membranes, is described here. High resolution solid-state NMR is deeply intertwined with the history of NMR, and the seminal paper was published in 1948. Although the general principles were understood by the end of the 1950s, it has taken more than fifty years for instrumentation and experimental methods to become equal to the technical problems presented by the biological assemblies of greatest interest. It is now possible to obtain atomic resolution structures of viral coat proteins in virus particles and membrane proteins in phospholipid bilayers by oriented sample solid-state NMR methods. The development of this aspect of the field of solid-state NMR is summarized in this review article. PMID:26069880

  13. Solid-State NMR Spectroscopy for the Physical Chemistry Laboratory

    ERIC Educational Resources Information Center

    Kinnun, Jacob J.; Leftin, Avigdor; Brown, Michael F.

    2013-01-01

    Solid-state nuclear magnetic resonance (NMR) spectroscopy finds growing application to inorganic and organic materials, biological samples, polymers, proteins, and cellular membranes. However, this technique is often neither included in laboratory curricula nor typically covered in undergraduate courses. On the other hand, spectroscopy and

  14. Solid-State NMR Spectroscopy for the Physical Chemistry Laboratory

    ERIC Educational Resources Information Center

    Kinnun, Jacob J.; Leftin, Avigdor; Brown, Michael F.

    2013-01-01

    Solid-state nuclear magnetic resonance (NMR) spectroscopy finds growing application to inorganic and organic materials, biological samples, polymers, proteins, and cellular membranes. However, this technique is often neither included in laboratory curricula nor typically covered in undergraduate courses. On the other hand, spectroscopy and…

  15. GFT projection NMR spectroscopy for proteins in the solid state

    PubMed Central

    Franks, W. Trent; Atreya, Hanudatta S.; Szyperski, Thomas

    2011-01-01

    Recording of four-dimensional (4D) spectra for proteins in the solid state has opened new avenues to obtain virtually complete resonance assignments and three-dimensional (3D) structures of proteins. As in solution state NMR, the sampling of three indirect dimensions leads per se to long minimal measurement time. Furthermore, artifact suppression in solid state NMR relies primarily on radio-frequency pulse phase cycling. For an n-step phase cycle, the minimal measurement times of both 3D and 4D spectra are increased n times. To tackle the associated ‘sampling problem’ and to avoid sampling limited data acquisition, solid state G-Matrix Fourier Transform (SS GFT) projection NMR is introduced to rapidly acquire 3D and 4D spectral information. Specifically, (4,3)D (HA)CANCOCX and (3,2)D (HACA)NCOCX were implemented and recorded for the 6 kDa protein GB1 within about 10% of the time required for acquiring the conventional congeners with the same maximal evolution times and spectral widths in the indirect dimensions. Spectral analysis was complemented by comparative analysis of expected spectral congestion in conventional and GFT NMR experiments, demonstrating that high spectral resolution of the GFT NMR experiments enables one to efficiently obtain nearly complete resonance assignments even for large proteins. PMID:21052779

  16. Advances in solid-state NMR of cellulose.

    PubMed

    Foston, Marcus

    2014-06-01

    Nuclear magnetic resonance (NMR) spectroscopy is a well-established analytical and enabling technology in biofuel research. Over the past few decades, lignocellulosic biomass and its conversion to supplement or displace non-renewable feedstocks has attracted increasing interest. The application of solid-state NMR spectroscopy has long been seen as an important tool in the study of cellulose and lignocellulose structure, biosynthesis, and deconstruction, especially considering the limited number of effective solvent systems and the significance of plant cell wall three-dimensional microstructure and component interaction to conversion yield and rate profiles. This article reviews common and recent applications of solid-state NMR spectroscopy methods that provide insight into the structural and dynamic processes of cellulose that control bulk properties and biofuel conversion. PMID:24590189

  17. On the solid-state NMR spectra of naproxen

    NASA Astrophysics Data System (ADS)

    Czernek, Jiří

    2015-01-01

    Two previous measurements of the 13C and 1H NMR isotropic chemical shifts in crystalline naproxen, which is an important pharmaceutical compound, are confronted with the results obtained from several theoretical approaches capable of the proper treatment of solid-phase effects. In the underlying geometrical optimizations, two crystal structures are considered. The agreement between the data sets is quantified, including an evaluation of the similarity between the experimental solid-state NMR spectra. The 13C-1H heteronuclear correlations are analyzed, and their various assignments are discussed employing the statistical treatment of the differences between the measured and theoretical isotropic chemical shifts.

  18. Solid state 33S NMR of inorganic sulfides.

    PubMed

    Wagler, Todd A; Daunch, William A; Rinaldi, Peter L; Palmer, Allen R

    2003-04-01

    Solid state 33S NMR spectra of a variety of inorganic sulfides have been obtained at magnetic field strengths of 4.7 and 17.6T. Spectra acquired with magic angle spinning show considerable improvements in sensitivity and resolution when compared with static spectra. Multiple factors are considered when analyzing the spectral line widths, including; magnetic field inhomogeneity, dipolar coupling, chemical shift anisotropy, chemical shift dispersion (CSD), T(2) relaxation, and quadrupolar coupling. Quadrupolar coupling was expected to be the dominant line broadening mechanism. However, for most of the samples CSD was the prevailing line broadening mechanism. Thus, for many of the metal sulfides studied at a high magnetic field strength, the line widths were actually larger than those observed in the spectra at low field. This is atypical in solid state 33S NMR. Solid state 33S spin-lattice (T(1)) and spin-spin (T(2)) relaxation rates were measured for the first time and are discussed. This information will be useful in future efforts to use 33S NMR in the compositional and structural analysis of sulfur containing materials. PMID:12713969

  19. A "special perspectives" issue: Recent achievements and new directions in biomolecular solid state NMR

    NASA Astrophysics Data System (ADS)

    Tycko, Robert

    2015-04-01

    Twenty years ago, applications of solid state nuclear magnetic resonance (NMR) methods to real problems involving biological systems or biological materials were few and far between. Starting in the 1980s, a small number of research groups had begun to explore the possibility of obtaining structural and dynamical information about peptides, proteins, and other biopolymers from solid state NMR spectra. Progress was initially slow due to the relatively primitive state of solid state NMR probes, spectrometers, sample preparation methods, and pulse sequence techniques, coupled with the small number of people contributing to this research area. By the early 1990s, with the advent of new ideas about pulse sequence techniques such as dipolar recoupling, improvements in techniques for orienting membrane proteins and in technology for magic-angle spinning (MAS), improvements in the capabilities of commercial NMR spectrometers, and general developments in multidimensional spectroscopy, it began to appear that biomolecular solid state NMR might have a viable future. It was not until 1993 that the annual number of publications in this area crept above twenty.

  20. Investigating albendazole desmotropes by solid-state NMR spectroscopy.

    PubMed

    Chattah, Ana K; Zhang, Rongchun; Mroue, Kamal H; Pfund, Laura Y; Longhi, Marcela R; Ramamoorthy, Ayyalusamy; Garnero, Claudia

    2015-03-01

    Characterization of the molecular structure and physicochemical solid-state properties of the solid forms of pharmaceutical compounds is a key requirement for successful commercialization as potential active ingredients in drug products. These properties can ultimately have a critical effect on the solubility and bioavailability of the final drug product. Here, the desmotropy of Albendazole forms I and II was investigated at the atomic level. Ultrafast magic angle spinning (MAS) solid-state nuclear magnetic resonance (NMR) spectroscopy, together with powder X-ray diffraction, thermal analysis, and Fourier transform infrared spectroscopy, were performed on polycrystalline samples of the two solids in order to fully characterize and distinguish the two forms. High-resolution one-dimensional (1)H, (13)C, and (15)N together with two-dimensional (1)H/(1)H single quantum-single quantum, (1)H/(1)H single quantum-double quantum, and (1)H/(13)C chemical shift correlation solid-state NMR experiments under MAS conditions were extensively used to decipher the intramolecular and intermolecular hydrogen bonding interactions present in both solid forms. These experiments enabled the unequivocal identification of the tautomers of each desmotrope. Our results also revealed that both solid forms may be described as dimeric structures, with different intermolecular hydrogen bonds connecting the tautomers in each dimer. PMID:25584993

  1. RNA structure determination by solid-state NMR spectroscopy

    PubMed Central

    Marchanka, Alexander; Simon, Bernd; Althoff-Ospelt, Gerhard; Carlomagno, Teresa

    2015-01-01

    Knowledge of the RNA three-dimensional structure, either in isolation or as part of RNP complexes, is fundamental to understand the mechanism of numerous cellular processes. Because of its flexibility, RNA represents a challenge for crystallization, while the large size of cellular complexes brings solution-state NMR to its limits. Here, we demonstrate an alternative approach on the basis of solid-state NMR spectroscopy. We develop a suite of experiments and RNA labeling schemes and demonstrate for the first time that ssNMR can yield a RNA structure at high-resolution. This methodology allows structural analysis of segmentally labelled RNA stretches in high-molecular weight cellular machinesindependent of their ability to crystallize and opens the way to mechanistic studies of currently difficult-to-access RNA-protein assemblies. PMID:25960310

  2. Solid-state 33S MAS NMR of inorganic sulfates.

    PubMed

    Wagler, Todd A; Daunch, William A; Panzner, Matthew; Youngs, Wiley J; Rinaldi, Peter L

    2004-10-01

    Solid-state (33)S MAS NMR spectra of a variety of inorganic sulfates have been obtained at magnetic field strengths of 4.7, 14.1, 17.6, and 18.8 T. Some of the difficulties associated with obtaining natural abundance (33)S NMR spectra have been overcome by using a high magnetic field strength and magic angle spinning (MAS). Multiple factors were considered when analyzing the spectral linewidths, including magnetic field inhomogeneity, dipolar coupling, chemical shift anisotropy, chemical shift dispersion, and quadrupolar coupling. In most of these sulfate samples, quadrupolar coupling was the dominant line broadening mechanism. Nuclear electric quadrupolar coupling constants (C(q)) as large as 2.05 MHz were calculated using spectral simulation software. Spectral information from these new data are compared with X-ray measurements and GAUSSIAN 98W calculations. A general correlation was observed between the magnitude of the C(q) and the increasing difference between S-O bond distances within the sulfate groups. Solid-state (33)S spin-lattice (T(1)) relaxation times were measured and show a significant reduction in T(1) for the hydrated sulfates. This is most likely the result of the modulation of the time-dependent electric field gradient at the nuclear site by motion of water molecules. This information will be useful in future efforts to use (33)S NMR in the compositional and structural analysis of sulfur containing materials. PMID:15388098

  3. Membrane protein structure determination using solid-state NMR.

    PubMed

    Watts, Anthony; Straus, Suzana K; Grage, Stephan L; Kamihira, Miya; Lam, Yuen Han; Zhao, Xin

    2004-01-01

    Solid-state NMR is emerging as a method for resolving structural information for large biomolecular complexes, such as membrane-embedded proteins. In principle, there is no molecular weight limit to the use of the approach, although the complexity and volume of data is still outside complete assignment and structural determinations for any large (Mr > approx 30,000) complex unless specific methods to reduce the information content to a manageable amount are employed. Such methods include specific residue-type labeling, labeling of putative segments of a protein, or examination of complexes made up of smaller, manageable units, such as oligomeric ion channels. Labeling possibilities are usually limited to recombinant or synthesized proteins, and labeling strategies often follow models from a bioinformatics approach. In all cases, and in common with most membrane studies, sample preparation is vital, and this activity alone can take considerable effort before NMR can be applied--peptide or protein production (synthesis or expression) followed by reconstitution into bilayers and resolution of suitable sample geometry is still technically challenging. As experience is gained in the field, this development time should decrease. Here, the practical aspects of the use of solid-state NMR for membrane protein structural determinations are presented, as well as how the methodology can be applied. Some successes to date are discussed, with an indication of how the area might develop. PMID:15318005

  4. Magic-angle spinning solid-state multinuclear NMR on low-field instrumentation

    NASA Astrophysics Data System (ADS)

    Srensen, Morten K.; Bakharev, Oleg; Jensen, Ole; Jakobsen, Hans J.; Skibsted, Jrgen; Nielsen, Niels Chr.

    2014-01-01

    Mobile and cost-effective NMR spectroscopy exploiting low-field permanent magnets is a field of tremendous development with obvious applications for arrayed large scale analysis, field work, and industrial screening. So far such demonstrations have concentrated on relaxation measurements and lately high-resolution liquid-state NMR applications. With high-resolution solid-state NMR spectroscopy being increasingly important in a broad variety of applications, we here introduce low-field magic-angle spinning (MAS) solid-state multinuclear NMR based on a commercial ACT 0.45 T 62 mm bore Halbach magnet along with a homebuilt FPGA digital NMR console, amplifiers, and a modified standard 45 mm wide MAS probe for 7 mm rotors. To illustrate the performance of the instrument and address cases where the low magnetic field may offer complementarity to high-field NMR experiments, we demonstrate applications for 23Na MAS NMR with enhanced second-order quadrupolar coupling effects and 31P MAS NMR where reduced influence from chemical shift anisotropy at low field may facilitate determination of heteronuclear dipole-dipole couplings.

  5. Solid-state NMR spectroscopy of protein complexes.

    PubMed

    Sun, Shangjin; Han, Yun; Paramasivam, Sivakumar; Yan, Si; Siglin, Amanda E; Williams, John C; Byeon, In-Ja L; Ahn, Jinwoo; Gronenborn, Angela M; Polenova, Tatyana

    2012-01-01

    Protein-protein interactions are vital for many biological processes. These interactions often result in the formation of protein assemblies that are large in size, insoluble, and difficult to crystallize, and therefore are challenging to study by structure biology techniques, such as single crystal X-ray diffraction and solution NMR spectroscopy. Solid-state NMR (SSNMR) spectroscopy is emerging as a promising technique for studies of such protein assemblies because it is not limited by molecular size, solubility, or lack of long-range order. In the past several years, we have applied magic angle spinning SSNMR-based methods to study several protein complexes. In this chapter, we discuss the general SSNMR methodologies employed for structural and dynamics analyses of protein complexes with specific examples from our work on thioredoxin reassemblies, HIV-1 capsid protein assemblies, and microtubule-associated protein assemblies. We present protocols for sample preparation and characterization, pulse sequences, SSNMR spectra collection, and data analysis. PMID:22167681

  6. Solid-state NMR characterization of Mowry Formation shales

    SciTech Connect

    Miknis, F.P.

    1992-04-01

    Solid-state [sup 13]C and [sup 29]Si NMR measurements were carried out on a series of petroleum source rocks from the Mowry Formation of the Powder River Basin in Wyoming. The objectives of this study wereto use CP/MAS [sup 13]C NMR measurements to monitor changes in the carbon structure of the kerogen that result from depth of burial, and to examine the feasibility of [sup 29]Si NMR for studying the thermal alteration of clay minerals during diagenesis. Carbon and silicon NMR measurements were made on a suite of samples covering a present-day depth interval of 3,000 to 11,500 ft.In general, the NMR results endorsed other geochemical analyses that were performed on the source rocks as part of another study to examine pressure compartmentalization in the Mowry Formation. The carbon aromaticity of the kerogen increased with depth of burial, and at depths greater that approximately 10,000 ft the kerogen showed little capacity to generate additional oil because of the small fraction of residual aliphatic carbon. By combining NMR and Rock-Eval measurements, an estimate of the hydrogen budget was obtained. The calculations indicated that approximately 20% of the kerogen was converted to hydrocarbons, and that sufficient hydrogen was liberated from aromatization and condensation reactions to stabilize the generated products. The [sup 29]Si NMR spectra were characterized by a relatively sharp quartz resonance and a broad resonance from the clay minerals. With increasing depth of burial, the clay resonance became broader and shifted slightly downfield. These changes qualitatively support X-ray analysis that shows progressive alteration of illite to smectite with depth of burial.

  7. Solid-state NMR characterization of Mowry Formation shales

    SciTech Connect

    Miknis, F.P.

    1992-04-01

    Solid-state {sup 13}C and {sup 29}Si NMR measurements were carried out on a series of petroleum source rocks from the Mowry Formation of the Powder River Basin in Wyoming. The objectives of this study wereto use CP/MAS {sup 13}C NMR measurements to monitor changes in the carbon structure of the kerogen that result from depth of burial, and to examine the feasibility of {sup 29}Si NMR for studying the thermal alteration of clay minerals during diagenesis. Carbon and silicon NMR measurements were made on a suite of samples covering a present-day depth interval of 3,000 to 11,500 ft.In general, the NMR results endorsed other geochemical analyses that were performed on the source rocks as part of another study to examine pressure compartmentalization in the Mowry Formation. The carbon aromaticity of the kerogen increased with depth of burial, and at depths greater that approximately 10,000 ft the kerogen showed little capacity to generate additional oil because of the small fraction of residual aliphatic carbon. By combining NMR and Rock-Eval measurements, an estimate of the hydrogen budget was obtained. The calculations indicated that approximately 20% of the kerogen was converted to hydrocarbons, and that sufficient hydrogen was liberated from aromatization and condensation reactions to stabilize the generated products. The {sup 29}Si NMR spectra were characterized by a relatively sharp quartz resonance and a broad resonance from the clay minerals. With increasing depth of burial, the clay resonance became broader and shifted slightly downfield. These changes qualitatively support X-ray analysis that shows progressive alteration of illite to smectite with depth of burial.

  8. Recrystallized S-layer protein of a probiotic Propionibacterium: structural and nanomechanical changes upon temperature or pH shifts probed by solid-state NMR and AFM.

    PubMed

    de sa Peixoto, Paulo; Roiland, Claire; Thomas, Daniel; Briard-Bion, Valérie; Le Guellec, Rozenn; Parayre, Sandrine; Deutsch, Stéphanie-Marie; Jan, Gwénaël; Guyomarc'h, Fanny

    2015-01-01

    Surface protein layers (S layers) are common constituents of the bacterial cell wall and originate from the assembly of strain-dependent surface layer proteins (Slps). These proteins are thought to play important roles in the bacteria's biology and to have very promising technological applications as biomaterials or as part of cell-host cross-talk in probiotic mechanism. The SlpA from Propionibacterium freudenreichii PFCIRM 118 strain was isolated and recrystallized to investigate organization and assembly of the protein using atomic force microscopy and solid-state (1)H and (13)C-nuclear magnetic resonance. SlpA was found to form hexagonal p1 monolayer lattices where the protein exhibited high proportions of disordered regions and of bound water. The lattice structure was maintained, but softened, upon mild heating or acidification, probably in relation with the increasing mobilities of the disordered protein regions. These results gave structural insights on the mobile protein regions exposed by S layer films, upon physiologically relevant changes of their environmental conditions. PMID:25479375

  9. Probing intermolecular hydrogen bonding in sibenadet hydrochloride polymorphs by high-resolution (1) H double-quantum solid-state NMR spectroscopy.

    PubMed

    Bradley, Jonathan P; Pickard, Chris J; Burley, Jonathan C; Martin, Dave R; Hughes, Leslie P; Cosgrove, Stephen D; Brown, Steven P

    2012-05-01

    Molecular packing in two polymorphs of sibenadet hydrochloride (AR-C68397AA, Viozan™) is investigated using a combined experimental (1) H double-quantum (DQ) solid-state magic-angle spinning nuclear magnetic resonance and computational (gauge including projected augmented wave calculation of chemical shifts) approach. For Form I, NH-NH and NH-OH (1) H DQ peaks are observed corresponding to nearest distances of 2.62 and 2.87 Å, respectively, for the intermolecular hydrogen-bonding arrangement in the single-crystal X-ray diffraction structure. The same (1) H DQ peaks at the same (1) H chemical shifts are observed for Form II, for which there is no single-crystal diffraction structure, indicating the same intermolecular hydrogen-bonding arrangement of the benzothiazolone moieties as in Form I. (1) H DQ build-up (as a function of the DQ recoupling time) curves are presented for the resolved NH-NH and NH-OH DQ peaks for the two polymorphs. For Form I, the ratio of the maximum intensity for the NH-OH and NH-NH DQ peaks is in excellent agreement with the ratio of the summed squares of the H-H dipolar couplings, as determined using H-H distances from the crystal structure up to 4 Å. Small differences in the (1) H DQ build-up behaviour for the two polymorphs are attributed to differences in the longer-range NH-OH distances associated with different inter-layer arrangements. PMID:22359321

  10. Probing the Role of Ceramide Headgroup Polarity in Short-Chain Model Skin Barrier Lipid Mixtures by (2)H Solid-State NMR Spectroscopy.

    PubMed

    Stahlberg, Sören; Lange, Stefan; Dobner, Bodo; Huster, Daniel

    2016-03-01

    The thermoptropic phase behaviors of two stratum corneum model lipid mixtures composed of equimolar contributions of either Cer[NS18] or Cer[NP18] with stearic acid and cholesterol were compared. Each component of the mixture was specifically deuterated such that the temperature-dependent (2)H NMR spectra allowed disentanglement of the complicated phase polymorphism of these lipid mixtures. While Cer[NS] is based on the sphingosine backbone, Cer[NP] features a phytosphingosine, which introduces an additional hydroxyl group into the headgroup of the ceramide and abolishes the double bond. From the NMR spectra, the individual contributions of all lipids to the respective phases could be determined. The comparison of the two lipid mixtures reveals that Cer[NP] containing mixtures have a tendency to form more fluid phases. It is concluded that the additional hydroxyl group of the phytosphingosine-containing ceramide Cer[NP18] in mixture with chain-matched stearic acid and cholesterol creates a packing defect that destabilizes the orthorhombic phase state of canonical SC mixtures. This steric clash favors the gel phase and promotes formation of fluid phases of Cer[NP] containing lipid mixtures at lower temperature compared to those containing Cer[NS18]. PMID:26828109

  11. Protein Dynamics in the Solid State from 2H NMR Line Shape Analysis. II. MOMD Applied to C–D and C–CD3 Probes

    PubMed Central

    2015-01-01

    Deuterium line shape analysis from mobile C–D and C–CD3 groups has emerged as a particularly useful tool for studying dynamics in the solid state. The theoretical models devised so far consist typically of sets of independent dynamic modes. Each such mode is simple and usually case-specific. In this scenario, model improvement entails adding yet another mode (thereby changing the overall model), comparison of different cases is difficult, and ambiguity is unavoidable. We recently developed the microscopic order macroscopic disorder (MOMD) approach as a single-mode alternative. In MOMD, the local spatial restrictions are expressed by an anisotropic potential, the local motion by a diffusion tensor, and the local molecular geometry by relative (magnetic and model-related) tensor orientations, all of adjustable symmetry. This approach provides a consistent method of analysis, thus resolving the issues above. In this study, we apply MOMD to PS-adsorbed LKα14 peptide and dimethylammonium tetraphenylborate (C–CD3 and N–CD3 dynamics, respectively), as well as HhaI methyltransferase target DNA and phase III of benzene-6-hexanoate (C–D dynamics). The success with fitting these four disparate cases, as well as the two cases in the previous report, demonstrates the generality of this MOMD-based approach. In this study, C–D and C–CD3 are both found to execute axial diffusion (rates R⊥ and R∥) in the presence of a rhombic potential given by the L = 2 spherical harmonics (coefficients c02 and c22). R⊥ (R∥) is in the 102–103 (104–105) s–1 range, and c02 and c22 are on the order of 2–3 kBT. Specific parameter values are determined for each mobile site. The diffusion and quadrupolar tensors are tilted at either 120° (consistent with trans–gauche isomerization) or nearly 110.5° (consistent with methyl exchange). Future prospects include extension of the MOMD formalism to include MAS, and application to 15N and 13C nuclei. PMID:26402431

  12. Solid State NMR Studies of the Aluminum Hydride Phases

    NASA Technical Reports Server (NTRS)

    Hwang, Son-Jong; Bowman, R. C., Jr.; Graetz, Jason; Reilly, J. J.

    2006-01-01

    Several solid state NMR techniques including magic-angle-spinning (MAS) and multiple-quantum (MQ) MAS experiments have been used to characterize various AlH3 samples. MAS-NMR spectra for the 1H and 27Al nuclei have been obtained on a variety of AlH3 samples that include the (beta)- and (gamma)- phases as well as the most stable (alpha)-phase. While the dominant components in these NMR spectra correspond to the aluminum hydride phases, other species were found that include Al metal, molecular hydrogen (H2), as well as peaks that can be assigned to Al-O species in different configurations. The occurrence and concentration of these extraneous components are dependent upon the initial AlH3 phase composition and preparation procedures. Both the (beta)-AlH3 and (gamma)-AlH3 phases were found to generate substantial amounts of Al metal when the materials were stored at room temperature while the (alpha)-phase materials do not exhibit these changes.

  13. Solid-State NMR Investigations of a Perfluorinated Ionomer (Nafion)

    NASA Astrophysics Data System (ADS)

    Chen, Qiang

    2005-03-01

    The chain dynamics and supramolecular structure of Nafion^, a perfluorinated ionomer which is widely used as a hydrophilic permselective membrane in fuel cells and chloralkali electrolysis, have been studied by solid-state NMR. With 1D and 2D NMR under 30-kHz magic-angle spinning (MAS), the ^19F and ^13C NMR peak widths and positions are determined, which corrects several previous assignments. The peak widths reveal static disorder around the branch point, increasing mobility towards the side group end, and a conformationally ordered backbone, which is essentially polytetrafluoroethylene (PTFE). Fast rotations of the helical backbone segments around their axis are confirmed in PTFE and observed similar in Nafion. The equal ^19F chemical shifts within parallel packed rotating chains in PTFE crystals result in slow ^19F spin diffusion between differently oriented chains. This spin diffusion is observed very fast for a majority of backbone segments in Nafion and the orientational correlations of the remainder backbones are weak. The typical diameter of backbone ``clusters'' in Nafion was 1 - 3 nm estimated by ^19F spin diffusion. Relatively fast ^19F spin exchange from any site in the side group to the backbone is observed. Absorbed water increases the side-group dynamics and conformational averaging, but not the segments near the branch point.

  14. In situ solid-state NMR for heterogeneous catalysis: a joint experimental and theoretical approach.

    PubMed

    Zhang, Weiping; Xu, Shutao; Han, Xiuwen; Bao, Xinhe

    2012-01-01

    In situ solid-state NMR is a well-established tool for investigations of the structures of the adsorbed reactants, intermediates and products on the surface of solid catalysts. The techniques allow identifications of both the active sites such as acidic sites and reaction processes after introduction of adsorbates and reactants inside an NMR rotor under magic angle spinning (MAS). The in situ solid-state NMR studies of the reactions can be achieved in two ways, i.e. under batch-like or continuous-flow conditions. The former technique is low cost and accessible to the commercial instrument while the latter one is close to the real catalytic reactions on the solids. This critical review describes the research progress on the in situ solid-state NMR techniques and the applications in heterogeneous catalysis under batch-like and continuous-flow conditions in recent years. Some typical probe molecules are summarized here to detect the Brønsted and Lewis acidic sites by MAS NMR. The catalytic reactions discussed in this review include methane aromatization, olefin selective oxidation and olefin metathesis on the metal oxide-containing zeolites. With combining the in situ MAS NMR spectroscopy and the density functional theoretical (DFT) calculations, the intermediates on the catalyst can be identified, and the reaction mechanism is revealed. Reaction kinetic analysis in the nanospace instead of in the bulk state can also be performed by employing laser-enhanced MAS NMR techniques in the in situ flow mode (163 references). PMID:21743940

  15. Molecular Structure of Humin and Melanoidin via Solid State NMR

    PubMed Central

    Herzfeld, Judith; Rand, Danielle; Matsuki, Yoh; Daviso, Eugenio; Mak-Jurkauskas, Melody; Mamajanov, Irena

    2011-01-01

    Sugar-derived humins and melanoidins figure significantly in food chemistry, agricultural chemistry, biochemistry and prebiotic chemistry. Despite wide interest and significant experimental attention, the amorphous and insoluble nature of the polymers has made them resistant to conventional structural characterization. Here we make use of solid-state NMR methods, including selective 13C substitution, 1H-dephasing, and double quantum filtration. The spectra, and their interpretation, are simplified by relying exclusively on hydronium for catalysis. The results for polymers derived from ribose, deoxyribose and fructose indicate diverse pathways to furans, suggest a simple route to pyrroles in the presence of amines, and reveal a heterogenous network-type polymer in which sugar molecules cross-link the heterocycles. PMID:21456563

  16. A Solid-State NMR Investigation of MQ Silicone Copolymers.

    PubMed

    Vasil'ev, Sergey G; Volkov, Vitaly I; Tatarinova, Elena A; Muzafarov, Aziz M

    2013-01-01

    The structure of MQ copolymers of the general chemical formula [(CH3)3SiO0.5]m [SiO2]n was characterized by means of solid-state magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. The MQ copolymers are highly branched polycyclic compounds (densely cross-linked nanosized networks). MQ copolymers were prepared by hydrolytic polycondensation in active medium. (29)Si NMR spectra were obtained by single pulse excitation (or direct polarization, DP) and cross-polarization (CP) (29)Si{(1)H} techniques in concert with MAS. It was shown that material consist of monofunctional M (≡SiO Si (CH3)3) and two types of tetrafunctional Q units: Q(4) ((≡SiO)4 Si) and Q(3) ((≡SiO)3 SiOH). Spin-lattice relaxation times T 1 measurements of (29)Si nuclei and analysis of (29)Si{(1)H} variable contact time signal intensities allowed us to obtain quantitative data on the relative content of different sites in copolymers. These investigations indicate that MQ copolymers represent dense structure with core and shell. PMID:23914072

  17. Probing hydrogen in ZnO nanorods using solid-state 1H nuclear magnetic resonance

    SciTech Connect

    Wang, Li Q.; Exarhos, Gregory J.; Windisch, Charles F.; Yao, Chunhua; Pederson, Larry R.; Zhou, Xiao Dong

    2007-04-23

    We have developed a low-temperature reflux method to synthesize large quantities of well-dispersed free-standing ZnO nanorods using a simple and mild aqueous solution route. In this approach, different surfactants were used to control nanostructure morphologies. Bound proton states in these ZnO nanorods were characterized for the first time by high resolution solid-state 1H magic angle spinning (MAS) NMR. In contrast to commercially available ZnO nano- or micro-particles, our uniform ZnO nanorods show a surprisingly sharp 1H NMR resonance. The feature is maintained upon heating to 500 oC, which suggests that an unusually stable proton species exists, most likely associated with lattice defects within the ZnO framework. Work here has demonstrated a new approach for probing a small amount of proton species associated with defects in nano-crystalline solids using high resolution solid-state 1H MAS NMR.

  18. 33S solid state NMR of sulphur speciation in silicate glasses.

    PubMed

    Couch, S; Howes, A P; Kohn, S C; Smith, M E

    2004-01-01

    33S solid-state NMR is reported from some model crystalline sulphides, sulphates, sulphites and thiosulphates. This is the first report of (33)S NMR signals of the latter two species from a solid. Good quality spectra, which have distinct, well separated shift ranges can be identified for all these groupings, except for sulphites and hydrogen sulphites whose resonances are very broad. Nonetheless the presence of sulphites and hydrogen sulphites can be confirmed from their characteristic sharp, intense time domain echoes. (33)S MAS NMR is also applied to a range of dry and hydrous silicate glasses with approximately 1wt% 99at% (33)S-enriched sulphur. It is clear that in all these glasses sulphate is present, and in mixed cation systems there is some evidence of preferential association of sulphate with a specific cation. In a dry potassium silicate glass sample two resolved (33)S resonances are observed, a sulphate, and the second from thiosulphate. Hence solid state (33)S NMR is shown to be a feasible probe that can be applied to this problem that can readily distinguish different sulphur species. PMID:15388185

  19. Structure determination of helical filaments by solid-state NMR spectroscopy.

    PubMed

    He, Lichun; Bardiaux, Benjamin; Ahmed, Mumdooh; Spehr, Johannes; König, Renate; Lünsdorf, Heinrich; Rand, Ulfert; Lührs, Thorsten; Ritter, Christiane

    2016-01-19

    The controlled formation of filamentous protein complexes plays a crucial role in many biological systems and represents an emerging paradigm in signal transduction. The mitochondrial antiviral signaling protein (MAVS) is a central signal transduction hub in innate immunity that is activated by a receptor-induced conversion into helical superstructures (filaments) assembled from its globular caspase activation and recruitment domain. Solid-state NMR (ssNMR) spectroscopy has become one of the most powerful techniques for atomic resolution structures of protein fibrils. However, for helical filaments, the determination of the correct symmetry parameters has remained a significant hurdle for any structural technique and could thus far not be precisely derived from ssNMR data. Here, we solved the atomic resolution structure of helical MAVS(CARD) filaments exclusively from ssNMR data. We present a generally applicable approach that systematically explores the helical symmetry space by efficient modeling of the helical structure restrained by interprotomer ssNMR distance restraints. Together with classical automated NMR structure calculation, this allowed us to faithfully determine the symmetry that defines the entire assembly. To validate our structure, we probed the protomer arrangement by solvent paramagnetic resonance enhancement, analysis of chemical shift differences relative to the solution NMR structure of the monomer, and mutagenesis. We provide detailed information on the atomic contacts that determine filament stability and describe mechanistic details on the formation of signaling-competent MAVS filaments from inactive monomers. PMID:26733681

  20. integrating Solid State NMR and Computations in Membrane Protein Science

    NASA Astrophysics Data System (ADS)

    Cross, Timothy

    2015-03-01

    Helical membrane protein structures are influenced by their native environment. Therefore the characterization of their structure in an environment that models as closely as possible their native environment is critical for achieving not only structural but functional understanding of these proteins. Solid state NMR spectroscopy in liquid crystalline lipid bilayers provides an excellent tool for such characterizations. Two classes of restraints can be obtained - absolute restraints that constrain the structure to a laboratory frame of reference when using uniformly oriented samples (approximately 1° of mosaic spread) and relative restraints that restrain one part of the structure with respect to another part such as torsional and distance restraints. Here, I will discuss unique restraints derived from uniformly oriented samples and the characterization of initial structures utilizing both restraint types, followed by restrained molecular dynamics refinement in the same lipid bilayer environment as that used for the experimental restraint collection. Protein examples will be taken from Influenza virus and Mycobacterium tuberculosis. When available comparisons of structures to those obtained using different membrane mimetic environments will be shown and the causes for structural distortions explained based on an understanding of membrane biophysics and its sophisticated influence on membrane proteins.

  1. Coherent and stochastic averaging in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Nevzorov, Alexander A.

    2014-12-01

    A new approach for calculating solid-state NMR lineshapes of uniaxially rotating membrane proteins under the magic-angle spinning conditions is presented. The use of stochastic Liouville equation (SLE) allows one to account for both coherent sample rotation and stochastic motional averaging of the spherical dipolar powder patterns by uniaxial diffusion of the spin-bearing molecules. The method is illustrated via simulations of the dipolar powder patterns of rigid samples under the MAS conditions, as well as the recent method of rotational alignment in the presence of both MAS and rotational diffusion under the conditions of dipolar recoupling. It has been found that it is computationally more advantageous to employ direct integration over a spherical grid rather than to use a full angular basis set for the SLE solution. Accuracy estimates for the bond angles measured from the recoupled amide 1H-15N dipolar powder patterns have been obtained at various rotational diffusion coefficients. It has been shown that the rotational alignment method is applicable to membrane proteins approximated as cylinders with radii of approximately 20 , for which uniaxial rotational diffusion within the bilayer is sufficiently fast and exceeds the rate 2 105 s-1.

  2. High resolution 1H solid state NMR studies of polyethyleneterephthalate

    NASA Astrophysics Data System (ADS)

    Cheung, T. T. P.; Gerstein, B. C.; Ryan, L. M.; Taylor, R. E.; Dybowski, D. R.

    1980-12-01

    Molecular motions and spatial properties of the solid polymer polyethyleneterephthalate have been investigated using high resolution 1H solid state NMR techniques. The longitudinal spin relaxation time T1ρ of protons (1H) in the rotating frame was measured for a spin locking field ranging from 5 to 20 G. The decay of the 1H magnetization indicated the existence of two distinct T1ρ's and their field dependence shows that they are associated with two mobile phases of the polymer. The 1H magnetization also relaxes under the dipolar narrowed Carr-Purcell (DNCP) multipulse sequence with two dintinct T1y relaxation times. The ratios T1y's and T1ρ's deviate significantly from the expected theoretical values. The combined experiment with magic angle spinning and the DNCP sequence followed by homonuclear dipolar decoupling reveals the individual T1y relaxation of the resolved methylene and aromatic protons. These two species of protons were found to relax with the same T1y's, thus implying that spin diffusion must have taken place under the homonuclear dipolar decoupling multipulse. The qualitative description of spin diffusion under homonuclear decoupling is given. The combined experiment with spin locking and the DNCP sequence yields the correspondence between the two T1ρ's and the two T1y's. The long T1ρ corresponds to the short T1y whereas the short T1ρ corresponds to the long T1y. Communication between the two spatial phases via spin diffusion was also observed in this experiment by monitoring the recovery of the 1H magnitization associated with the short T1ρ after it has been eliminated during the spin locking. The total 1H magnetization is allowed to equilibrate in the laboratory frame for a variable time much shorter than T1 after the spin locking field has been turned off. The spatial relationship between the two phases is discussed.

  3. Arabidopsis thalianafrom Polarization Transfer Solid-State NMR

    SciTech Connect

    White, Paul B; Wang, Tuo; Park, Yong Bum; Cosgrove, Daniel J; Hong, Mei

    2014-07-23

    Polysaccharide-rich plant cell walls are hydrated under functional conditions, but the molecular interactions between water and polysaccharides in the wall have not been investigated. In this work, we employ polarization transfer solid-state NMR techniques to study the hydration of primary-wall polysaccharides of the model plant, Arabidopsis thaliana. By transferring water 1H polarization to polysaccharides through distance- and mobility-dependent 1H–1H dipolar couplings and detecting it through polysaccharide 13C signals, we obtain information about water proximity to cellulose, hemicellulose, and pectins as well as water mobility. Both intact and partially extracted cell wall samples are studied. Our results show that water–pectin polarization transfer is much faster than water–cellulose polarization transfer in all samples, but the extent of extraction has a profound impact on the water–polysaccharide spin diffusion. Removal of calcium ions and the consequent extraction of homogalacturonan (HG) significantly slowed down spin diffusion, while further extraction of matrix polysaccharides restored the spin diffusion rate. These trends are observed in cell walls with similar water content, thus they reflect inherent differences in the mobility and spatial distribution of water. Combined with quantitative analysis of the polysaccharide contents, our results indicate that calcium ions and HG gelation increase the amount of bound water, which facilitates spin diffusion, while calcium removal disrupts the gel and gives rise to highly dynamic water, which slows down spin diffusion. The recovery of spin diffusion rates after more extensive extraction is attributed to increased water-exposed surface areas of the polysaccharides. Water–pectin spin diffusion precedes water–cellulose spin diffusion, lending support to the single-network model of plant primary walls in which a substantial fraction of the cellulose surface is surrounded by pectins.

  4. Structural studies of proteins by paramagnetic solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Jaroniec, Christopher P.

    2015-04-01

    Paramagnetism-based nuclear pseudocontact shifts and spin relaxation enhancements contain a wealth of information in solid-state NMR spectra about electron-nucleus distances on the ∼20 Å length scale, far beyond that normally probed through measurements of nuclear dipolar couplings. Such data are especially vital in the context of structural studies of proteins and other biological molecules that suffer from a sparse number of experimentally-accessible atomic distances constraining their three-dimensional fold or intermolecular interactions. This perspective provides a brief overview of the recent developments and applications of paramagnetic magic-angle spinning NMR to biological systems, with primary focus on the investigations of metalloproteins and natively diamagnetic proteins modified with covalent paramagnetic tags.

  5. Recent technique developments and applications of solid state NMR in characterising inorganic materials.

    PubMed

    Hanna, J V; Smith, M E

    2010-07-01

    A broad overview is given of some key recent developments in solid state NMR techniques that have driven enhanced applications to inorganic materials science. Reference is made to advances in hardware, pulse sequences and associated computational methods (e.g. first principles calculations, spectral simulation), along with their combination to provide more information about solid phases. The resulting methodology has allowed more nuclei to be observed and more structural information to be extracted. Cross referencing between experimental parameters and their calculation from the structure has given an added dimension to NMR as a characterisation probe of materials. Emphasis is placed on the progress made in the last decade especially from those nuclei that were little studied previously. The general points about technique development and the increased range of nuclei observed are illustrated through some specific exemplars from inorganic materials science. PMID:20605082

  6. Synthesis and solid state NMR characterization of novel peptide/silica hybrid materials.

    PubMed

    Werner, Mayke; Heil, Andreas; Rothermel, Niels; Breitzke, Hergen; Groszewicz, Pedro Braga; Thankamony, Aany Sofia; Gutmann, Torsten; Buntkowsky, Gerd

    2015-11-01

    The successful synthesis and solid state NMR characterization of silica-based organic-inorganic hybrid materials is presented. For this, collagen-like peptides are immobilized on carboxylate functionalized mesoporous silica (COOH/SiOx) materials. A pre-activation of the silica material with TSTU (O-(N-Succinimidyl)-N,N,N',N'-tetramethyluronium tetrafluoroborate) is performed to enable a covalent binding of the peptides to the linker. The success of the covalent immobilization is indicated by the decrease of the (13)C CP-MAS NMR signal of the TSTU moiety. A qualitative distinction between covalently bound and adsorbed peptide is feasible by (15)N CP-MAS Dynamic Nuclear Polarization (DNP). The low-field shift of the (15)N signal of the peptide's N-terminus clearly identifies it as the binding site. The DNP enhancement allows the probing of natural abundance (15)N nuclei, rendering expensive labeling of peptides unnecessary. PMID:26411982

  7. Preparation of RNA samples with narrow line widths for solid state NMR investigations

    NASA Astrophysics Data System (ADS)

    Huang, Wei; Bardaro, Michael F.; Varani, Gabriele; Drobny, Gary P.

    2012-10-01

    Solid state NMR can provide detailed structural and dynamic information on biological systems that cannot be studied under solution conditions, and can investigate motions which occur with rates that cannot be fully studied by solution NMR. This approach has successfully been used to study proteins, but the application of multidimensional solid state NMR to RNA has been limited because reported line widths have been too broad to execute most multidimensional experiments successfully. A reliable method to generate spectra with narrow line widths is necessary to apply the full range of solid state NMR spectroscopic approaches to RNA. Using the HIV-1 transactivation response (TAR) RNA as a model, we present an approach based on precipitation with polyethylene glycol that improves the line width of 13C signals in TAR from >6 ppm to about 1 ppm, making solid state 2D NMR studies of selectively enriched RNAs feasible at ambient temperature.

  8. Solid-State NMR Examination of Alteration Layers on a Nuclear Waste Glasses

    SciTech Connect

    Murphy, Kelly A.; Washton, Nancy M.; Ryan, Joseph V.; Pantano, Carlo G.; Mueller, Karl T.

    2013-06-01

    Solid-state NMR is a powerful tool for probing the role and significance of alteration layers in determining the kinetics for the corrosion of nuclear waste glass. NMR methods are used to probe the chemical structure of the alteration layers to elucidate information about their chemical complexity, leading to increased insight into the mechanism of altered layer formation. Two glass compositions were examined in this study: a glass preliminarily designed for nuclear waste immobilization (called AFCI) and a simplified version of this AFCI glass (which we call SA1R). Powdered glasses with controlled and known particles sizes were corroded at 90 °C for periods of one and five months with a surface-area to solution-volume ratio of 100,000 m-1. 1H-29Si CP-CPMG MAS NMR, 1H-27Al CP-MAS NMR, 1H-11B CP-MAS NMR, and 1H-23Na CP-MAS NMR experiments provide isolated structural information about the alteration layers, which differ in structure from that of the pristine glass. Both glasses studied here develop alteration layers composed primarily of [IV]Si species. Aluminum is also retained in the alteration layers, perhaps facilitated by the observed increase in coordination from [IV]Al to [VI]Al, which correlates with a loss of charge balancing cations. 1H-11B CP-MAS NMR observations indicated a retention of boron in hydrated glass layers, which has not been characterized by previous work. For the AFCI glass, secondary phase formation begins during the corrosion times considered here, and these neophases are detected within the alteration layers. We identify precursor phases as crystalline sodium metasilicates. An important finding is that layer thickness depends on the length of the initial alteration stages and varies only with respect to silicon species during the residual rate regime.

  9. Solid-state NMR studies of biomineralization peptides and proteins.

    PubMed

    Roehrich, Adrienne; Drobny, Gary

    2013-09-17

    Nature has evolved sophisticated strategies for engineering hard tissues through the interaction of proteins, and ultimately cells, with inorganic mineral phases. This process, called biomineralization, is how living organisms transform inorganic materials such as hydroxyapatite, calcite, and silica into highly intricate and organized structures. The remarkable material properties of shell, bone, and teeth come from the activities of proteins that function at the organic-inorganic interface. A better understanding of the biomolecular mechanisms used to promote or retard the formation of mineral-based structures could provide important design principles for the development of calcification inhibitors and promoters in orthopedics, cardiology, urology, and dentistry. With the knowledge of the structural basis for control of hard tissue growth by proteins, scientists could potentially develop materials using biomimetic principles with applications in catalysis, biosensors, electronic devices, and chromatographic separations, to name a few. Additionally, biomineralization also has potential applications in electronics, catalysis, magnetism, sensory devices, and mechanical design. Where man-made hard materials require the use of extreme temperatures, high pressure, and pH, biological organisms can accomplish these feats at ambient temperature and at physiological pH. Despite the fact that many researchers want to identify and control the structure of proteins at material and biomineral interfaces, there is a decided lack of molecular-level structure information available for proteins at biomaterial interfaces in general. In particular, this holds for mammalian proteins that directly control calcification processes in hard tissue. The most fundamental questions regarding the secondary and tertiary structures of proteins adsorbed to material surfaces, how proteins catalyze the formation of biomineral composites, or how proteins interact at biomaterial interfaces remain unanswered. This is largely due to a lack of methods capable of providing high-resolution structural information for proteins adsorbed to material surfaces under physiologically relevant conditions. In this Account, we highlight recent work that is providing insight into the structure and crystal recognition mechanisms of a salivary protein model system, as well as the structure and interactions of a peptide that catalyzes the formation of biosilica composites. To develop a better understanding of the structure and interactions of proteins in biomaterials, we have used solid-state NMR techniques to determine the molecular structure and dynamics of proteins and peptides adsorbed onto inorganic crystal surfaces and embedded within biomineral composites. This work adds to the understanding of the structure and crystal recognition mechanisms of an acidic human salivary phosphoprotein, statherin. PMID:23932180

  10. QUANTITATIVE SOLID-STATE 13C NMR SPECTROSCOPY OF ORGANIC MATTER FRACTIONS IN LOWLAND RICE SOILS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spin counting on solid-state **13C cross-polarization (CP) nuclear magnetic resonance (NMR) spectra of two humic fractions isolated from tropical lowland soils showed that only 32-81% of potential **13C NMR signal was detected. The observability of **13C NMR signal (Cobs) was higher in the mobile h...

  11. Solid State FT-IR and (31)P NMR Spectral Features of Phosphate Compounds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Solid-state spectroscopic techniques, including Fourier transform infrared (FT-IR) and solid-state 31P magic angle spinning nuclear magnetic resonance (MAS NMR) spectroscopies, are powerful tools for evaluating metal speciation and transformation mechanisms of P compounds in the environment. Studie...

  12. Analysis of trivalent cation complexation to functionalized mesoporous silica using solid-state NMR spectroscopy.

    PubMed

    Shusterman, Jennifer; Mason, Harris; Bruchet, Anthony; Zavarin, Mavrik; Kersting, Annie B; Nitsche, Heino

    2014-11-28

    Functionalized mesoporous silica has applications in separations science, catalysis, and sensors. In this work, we studied the fundamental interactions of trivalent cations with functionalized mesoporous silica. We contacted trivalent cations of varying ionic radii with N-[5-(trimethoxysilyl)-2-aza-1-oxopentyl]caprolactam functionalized mesoporous silica with the aim of probing the binding mechanism of the metal to the surface of the solid. We studied the functionalized silica using solid-state nuclear magnetic resonance (NMR) spectroscopy before and after contact with the metals of interest. We collected NMR spectra of the various metals, as well as of (29)Si and (13)C to probe the silica substrate and the ligand properties, respectively. The NMR spectra indicate that the metals bind to the functionalized silica via two mechanisms. Aluminum sorbed to both the silica and the ligand, but with different coordination for each. Scandium also sorbed to both the silica and the ligand, and unlike the aluminum, had the same coordination number. Additionally, the functionalized silica was susceptible to acid hydrolysis and two primary mechanisms of degradation were observed: detachment from the silica surface and opening of the seven-membered ring in the ligand. Opening of the seven-membered ring may be beneficial in that it decreases steric hindrance of the molecule for binding. PMID:25265419

  13. Optimized multiple quantum MAS lineshape simulations in solid state NMR

    NASA Astrophysics Data System (ADS)

    Brouwer, William J.; Davis, Michael C.; Mueller, Karl T.

    2009-10-01

    The majority of nuclei available for study in solid state Nuclear Magnetic Resonance have half-integer spin I>1/2, with corresponding electric quadrupole moment. As such, they may couple with a surrounding electric field gradient. This effect introduces anisotropic line broadening to spectra, arising from distinct chemical species within polycrystalline solids. In Multiple Quantum Magic Angle Spinning (MQMAS) experiments, a second frequency dimension is created, devoid of quadrupolar anisotropy. As a result, the center of gravity of peaks in the high resolution dimension is a function of isotropic second order quadrupole and chemical shift alone. However, for complex materials, these parameters take on a stochastic nature due in turn to structural and chemical disorder. Lineshapes may still overlap in the isotropic dimension, complicating the task of assignment and interpretation. A distributed computational approach is presented here which permits simulation of the two-dimensional MQMAS spectrum, generated by random variates from model distributions of isotropic chemical and quadrupole shifts. Owing to the non-convex nature of the residual sum of squares (RSS) function between experimental and simulated spectra, simulated annealing is used to optimize the simulation parameters. In this manner, local chemical environments for disordered materials may be characterized, and via a re-sampling approach, error estimates for parameters produced. Program summaryProgram title: mqmasOPT Catalogue identifier: AEEC_v1_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/AEEC_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 3650 No. of bytes in distributed program, including test data, etc.: 73 853 Distribution format: tar.gz Programming language: C, OCTAVE Computer: UNIX/Linux Operating system: UNIX/Linux Has the code been vectorised or parallelized?: Yes RAM: Example: (1597 powder angles) × (200 Samples) × (81 F2 frequency pts) × (31 F1 frequency points) = 3.5M, SMP AMD opteron Classification: 2.3 External routines: OCTAVE ( http://www.gnu.org/software/octave/), GNU Scientific Library ( http://www.gnu.org/software/gsl/), OPENMP ( http://openmp.org/wp/) Nature of problem: The optimal simulation and modeling of multiple quantum magic angle spinning NMR spectra, for general systems, especially those with mild to significant disorder. The approach outlined and implemented in C and OCTAVE also produces model parameter error estimates. Solution method: A model for each distinct chemical site is first proposed, for the individual contribution of crystallite orientations to the spectrum. This model is averaged over all powder angles [1], as well as the (stochastic) parameters; isotropic chemical shift and quadrupole coupling constant. The latter is accomplished via sampling from a bi-variate Gaussian distribution, using the Box-Muller algorithm to transform Sobol (quasi) random numbers [2]. A simulated annealing optimization is performed, and finally the non-linear jackknife [3] is applied in developing model parameter error estimates. Additional comments: The distribution contains a script, mqmasOpt.m, which runs in the OCTAVE language workspace. Running time: Example: (1597 powder angles) × (200 Samples) × (81 F2 frequency pts) × (31 F1 frequency points) = 58.35 seconds, SMP AMD opteron. References:S.K. Zaremba, Annali di Matematica Pura ed Applicata 73 (1966) 293. H. Niederreiter, Random Number Generation and Quasi-Monte Carlo Methods, SIAM, 1992. T. Fox, D. Hinkley, K. Larntz, Technometrics 22 (1980) 29.

  14. Solid state NMR: The essential technology for helical membrane protein structural characterization

    NASA Astrophysics Data System (ADS)

    Cross, Timothy A.; Ekanayake, Vindana; Paulino, Joana; Wright, Anna

    2014-02-01

    NMR spectroscopy of helical membrane proteins has been very challenging on multiple fronts. The expression and purification of these proteins while maintaining functionality has consumed countless graduate student hours. Sample preparations have depended on whether solution or solid-state NMR spectroscopy was to be performed - neither have been easy. In recent years it has become increasingly apparent that membrane mimic environments influence the structural result. Indeed, in these recent years we have rediscovered that Nobel laureate, Christian Anfinsen, did not say that protein structure was exclusively dictated by the amino acid sequence, but rather by the sequence in a given environment (Anfinsen, 1973) [106]. The environment matters, molecular interactions with the membrane environment are significant and many examples of distorted, non-native membrane protein structures have recently been documented in the literature. However, solid-state NMR structures of helical membrane proteins in proteoliposomes and bilayers are proving to be native structures that permit a high resolution characterization of their functional states. Indeed, solid-state NMR is uniquely able to characterize helical membrane protein structures in lipid environments without detergents. Recent progress in expression, purification, reconstitution, sample preparation and in the solid-state NMR spectroscopy of both oriented samples and magic angle spinning samples has demonstrated that helical membrane protein structures can be achieved in a timely fashion. Indeed, this is a spectacular opportunity for the NMR community to have a major impact on biomedical research through the solid-state NMR spectroscopy of these proteins.

  15. Solid state NMR: The essential technology for helical membrane protein structural characterization

    PubMed Central

    Cross, Timothy A.; Ekanayake, Vindana; Paulino, Joana; Wright, Anna

    2014-01-01

    NMR spectroscopy of helical membrane proteins has been very challenging on multiple fronts. The expression and purification of these proteins while maintaining functionality has consumed countless graduate student hours. Sample preparations have depended on whether solution or solid-state NMR spectroscopy was to be performed – neither have been easy. In recent years it has become increasingly apparent that membrane mimic environments influence the structural result. Indeed, in these recent years we have rediscovered that Nobel laureate, Christian Anfinsen, did not say that protein structure was exclusively dictated by the amino acid sequence, but rather by the sequence in a given environment (Anfinsen, 1973) [106]. The environment matters, molecular interactions with the membrane environment are significant and many examples of distorted, non-native membrane protein structures have recently been documented in the literature. However, solid-state NMR structures of helical membrane proteins in proteoliposomes and bilayers are proving to be native structures that permit a high resolution characterization of their functional states. Indeed, solid-state NMR is uniquely able to characterize helical membrane protein structures in lipid environments without detergents. Recent progress in expression, purification, reconstitution, sample preparation and in the solid-state NMR spectroscopy of both oriented samples and magic angle spinning samples has demonstrated that helical membrane protein structures can be achieved in a timely fashion. Indeed, this is a spectacular opportunity for the NMR community to have a major impact on biomedical research through the solid-state NMR spectroscopy of these proteins. PMID:24412099

  16. Solid state NMR and LVSEM studies on the hardening of latex modified tile mortar systems

    SciTech Connect

    Rottstegge, J.; Arnold, M.; Herschke, L.; Glasser, G.; Wilhelm, M.; Spiess, H.W. . E-mail: spiess@mpip-mainz.mpg.de; Hergeth, W.D.

    2005-12-15

    Construction mortars contain a broad variety of both inorganic and organic additives beside the cement powder. Here we present a study of tile mortar systems based on portland cement, quartz, methyl cellulose and different latex additives. As known, the methyl cellulose stabilizes the freshly prepared cement paste, the latex additive enhances final hydrophobicity, flexibility and adhesion. Measurements were performed by solid state nuclear magnetic resonance (NMR) and low voltage scanning electron microscopy (LVSEM) to probe the influence of the latex additives on the hydration, hardening and the final tile mortar properties. While solid state NMR enables monitoring of the bulk composition, scanning electron microscopy affords visualization of particles and textures with respect to their shape and the distribution of the different phases. Within the alkaline cement paste, the poly(vinyl acetate) (VAc)-based latex dispersions stabilized by poly(vinyl alcohol) (PVA) were found to be relatively stable against hydrolysis. The influence of the combined organic additives methyl cellulose, poly(vinyl alcohol) and latexes stabilized by poly(vinyl alcohol) on the final silicate structure of the cement hydration products is small. But even small amounts of additives result in an increased ratio of ettringite to monosulfate within the final hydrated tile mortar as monitored by {sup 27}Al NMR. The latex was found to be adsorbed to the inorganic surfaces, acting as glue to the inorganic components. For similar latex water interfaces built up by poly(vinyl alcohol), a variation in the latex polymer composition results in modified organic textures. In addition to the networks of the inorganic cement and of the latex, there is a weak network build up by thin polymer fibers, most probably originating from poly(vinyl alcohol). Besides the weak network, polymer fibers form well-ordered textures covering inorganic crystals such as portlandite.

  17. A Solid-State NMR Experiment: Analysis of Local Structural Environments in Phosphate Glasses

    ERIC Educational Resources Information Center

    Anderson, Stanley E.; Saiki, David; Eckert, Hellmut; Meise-Gresch, Karin

    2004-01-01

    An experiment that can be used to directly study the local chemical environments of phosphorus in solid amorphous materials is demonstrated. The experiment aims at familiarizing the students of chemistry with the principles of solid-state NMR, by having them synthesize a simple phosphate glass, and making them observe the (super 31)P NMR spectrum,…

  18. CHARACTERIZATION OF SOIL ORGANIC MATTER IN TROPICAL RICE SOILS BY ADVANCED SOLID-STATE NMR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The chemical structures of soil organic matter in a continually submerged, triple-cropped lowland rice soil having large inputs of rice straw and an aerobic dryland rice soil were compared using advanced solid-state nuclear magnetic resonance (NMR) techniques. Quantitative 13C NMR shows that a humi...

  19. Heat management strategies for solid-state NMR of functional proteins.

    PubMed

    Fowler, Daniel J; Harris, Michael J; Thompson, Lynmarie K

    2012-09-01

    Modern solid-state NMR methods can acquire high-resolution protein spectra for structure determination. However, these methods use rapid sample spinning and intense decoupling fields that can heat and denature the protein being studied. Here we present a strategy to avoid destroying valuable samples. We advocate first creating a sacrificial sample, which contains unlabeled protein (or no protein) in buffer conditions similar to the intended sample. This sample is then doped with the chemical shift thermometer Sm2Sn2O7. We introduce a pulse scheme called TCUP (for Temperature Calibration Under Pulseload) that can characterize the heating of this sacrificial sample rapidly, under a variety of experimental conditions, and with high temporal resolution. Sample heating is discussed with respect to different instrumental variables such as spinning speed, decoupling strength and duration, and cooling gas flow rate. The effects of different sample preparation variables are also discussed, including ionic strength, the inclusion of cryoprotectants, and the physical state of the sample (i.e. liquid, solid, or slurry). Lastly, we discuss probe detuning as a measure of sample thawing that does not require retuning the probe or using chemical shift thermometer compounds. Use of detuning tests and chemical shift thermometers with representative sample conditions makes it possible to maximize the efficiency of the NMR experiment while retaining a functional sample. PMID:22868258

  20. Heat management strategies for solid-state NMR of functional proteins

    NASA Astrophysics Data System (ADS)

    Fowler, Daniel J.; Harris, Michael J.; Thompson, Lynmarie K.

    2012-09-01

    Modern solid-state NMR methods can acquire high-resolution protein spectra for structure determination. However, these methods use rapid sample spinning and intense decoupling fields that can heat and denature the protein being studied. Here we present a strategy to avoid destroying valuable samples. We advocate first creating a sacrificial sample, which contains unlabeled protein (or no protein) in buffer conditions similar to the intended sample. This sample is then doped with the chemical shift thermometer Sm2Sn2O7. We introduce a pulse scheme called TCUP (for Temperature Calibration Under Pulseload) that can characterize the heating of this sacrificial sample rapidly, under a variety of experimental conditions, and with high temporal resolution. Sample heating is discussed with respect to different instrumental variables such as spinning speed, decoupling strength and duration, and cooling gas flow rate. The effects of different sample preparation variables are also discussed, including ionic strength, the inclusion of cryoprotectants, and the physical state of the sample (i.e. liquid, solid, or slurry). Lastly, we discuss probe detuning as a measure of sample thawing that does not require retuning the probe or using chemical shift thermometer compounds. Use of detuning tests and chemical shift thermometers with representative sample conditions makes it possible to maximize the efficiency of the NMR experiment while retaining a functional sample.

  1. Heat Management Strategies for Solid-state NMR of Functional Proteins

    PubMed Central

    Fowler, Daniel J.; Harris, Michael J.; Thompson, Lynmarie K.

    2012-01-01

    Modern solid-state NMR methods can acquire high-resolution protein spectra for structure determination. However, these methods use rapid sample spinning and intense decoupling fields that can heat and denature the protein being studied. Here we present a strategy to avoid destroying valuable samples. We advocate first creating a sacrificial sample, which contains unlabeled protein (or no protein) in buffer conditions similar to the intended sample. This sample is then doped with the chemical shift thermometer Sm2Sn2O7. We introduce a pulse scheme called TCUP (for Temperature Calibration Under Pulseload) that can characterize the heating of this sacrificial sample rapidly, under a variety of experimental conditions, and with high temporal resolution. Sample heating is discussed with respect to different instrumental variables such as spinning speed, decoupling strength and duration, and cooling gas flow rate. The effects of different sample preparation variables are also discussed, including ionic strength, the inclusion of cryoprotectants, and the physical state of the sample (i.e. liquid, solid, or slurry). Lastly, we discuss probe detuning as a measure of sample thawing that does not require retuning the probe or using chemical shift thermometer compounds. Use of detuning tests and chemical shift thermometers with representative sample conditions makes it possible to maximize the efficiency of the NMR experiment while retaining a functional sample. PMID:22868258

  2. Detecting substrates bound to the secondary multidrug efflux pump EmrE by DNP-enhanced solid-state NMR.

    PubMed

    Ong, Yean Sin; Lakatos, Andrea; Becker-Baldus, Johanna; Pos, Klaas M; Glaubitz, Clemens

    2013-10-23

    Escherichia coli EmrE, a homodimeric multidrug antiporter, has been suggested to offer a convenient paradigm for secondary transporters due to its small size. It contains four transmembrane helices and forms a functional dimer. We have probed the specific binding of substrates TPP(+) and MTP(+) to EmrE reconstituted into 1,2-dimyristoyl-sn-glycero-3-phosphocholine liposomes by (31)P MAS NMR. Our NMR data show that both substrates occupy the same binding pocket but also indicate some degree of heterogeneity of the bound ligand population, reflecting the promiscuous nature of ligand binding by multidrug efflux pumps. Direct interaction between (13)C-labeled TPP(+) and key residues within the EmrE dimer has been probed by through-space (13)C-(13)C correlation spectroscopy. This was made possible by the use of solid-state NMR enhanced by dynamic nuclear polarization (DNP) through which a 19-fold signal enhancement was achieved. Our data provide clear evidence for the long assumed direct interaction between substrates such as TPP(+) and the essential residue E14 in transmembrane helix 1. Our work also demonstrates the power of DNP-enhanced solid-state NMR at low temperatures for the study for secondary transporters, which are highly challenging for conventional NMR detection. PMID:24047229

  3. In situ solid-state NMR spectroscopy of electrochemical cells: batteries, supercapacitors, and fuel cells.

    PubMed

    Blanc, Frédéric; Leskes, Michal; Grey, Clare P

    2013-09-17

    Electrochemical cells, in the form of batteries (or supercapacitors) and fuel cells, are efficient devices for energy storage and conversion. These devices show considerable promise for use in portable and static devices to power electronics and various modes of transport and to produce and store electricity both locally and on the grid. For example, high power and energy density lithium-ion batteries are being developed for use in hybrid electric vehicles where they improve the efficiency of fuel use and help to reduce greenhouse gas emissions. To gain insight into the chemical reactions involving the multiple components (electrodes, electrolytes, interfaces) in the electrochemical cells and to determine how cells operate and how they fail, researchers ideally should employ techniques that allow real-time characterization of the behavior of the cells under operating conditions. This Account reviews the recent use of in situ solid-state NMR spectroscopy, a technique that probes local structure and dynamics, to study these devices. In situ NMR studies of lithium-ion batteries are performed on the entire battery, by using a coin cell design, a flat sealed plastic bag, or a cylindrical cell. The battery is placed inside the NMR coil, leads are connected to a potentiostat, and the NMR spectra are recorded as a function of state of charge. (7)Li is used for many of these experiments because of its high sensitivity, straightforward spectral interpretation, and relevance to these devices. For example, (7)Li spectroscopy was used to detect intermediates formed during electrochemical cycling such as LixC and LiySiz species in batteries with carbon and silicon anodes, respectively. It was also used to observe and quantify the formation and growth of metallic lithium microstructures, which can cause short circuits and battery failure. This approach can be utilized to identify conditions that promote dendrite formation and whether different electrolytes and additives can help prevent dendrite formation. The in situ method was also applied to monitor (by (11)B NMR) electrochemical double-layer formation in supercapacitors in real time. Though this method is useful, it comes with challenges. The separation of the contributions from the different cell components in the NMR spectra is not trivial because of overlapping resonances. In addition, orientation-dependent NMR interactions, including the spatial- and orientation-dependent bulk magnetic susceptibility (BMS) effects, can lead to resonance broadening. Efforts to understand and mitigate these BMS effects are discussed in this Account. The in situ NMR investigation of fuel cells initially focused on the surface electrochemistry at the electrodes and the electrochemical oxidation of methanol and CO to CO2 on the Pt cathode. On the basis of the (13)C and (195)Pt NMR spectra of the adsorbates and electrodes, CO adsorbed on Pt and other reaction intermediates and complete oxidation products were detected and their mode of binding to the electrodes investigated. Appropriate design and engineering of the NMR hardware has allowed researchers to integrate intact direct methanol fuel cells into NMR probes. Chemical transformations of the circulating methanol could be followed and reaction intermediates could be detected in real time by either (2)H or (13)C NMR spectroscopy. By use of the in situ NMR approach, factors that control fuel cell performance, such as methanol cross over and catalyst performance, were identified. PMID:24041242

  4. Solid-state NMR studies of Ziegler-Natta and metallocene catalysts.

    PubMed

    Tijssen, Koen C H; Blaakmeer, E S Merijn; Kentgens, Arno P M

    2015-01-01

    Ziegler-Natta catalysts are the workhorses of polyolefin production. However, although they have been used and intensively studied for half a century, there is still no comprehensive picture of their mechanistic operation. New techniques are needed to gain more insight in these catalysts. Solid-state NMR has reached a high level of sophistication over the last few decades and holds great promise for providing a deeper insight in Ziegler-Natta catalysis. This review outlines the possibilities for solid-state NMR to characterize the different components and interactions in Ziegler-Natta and metallocene catalysts. An overview is given of some of the expected mechanisms and the resulting polymer microstructure and other characteristics. In the second part of this review we present studies that have used solid-state NMR to investigate the composition of Ziegler-Natta and metallocene catalysts or the interactions between their components. PMID:25957882

  5. Solid-state NMR studies of the dynamics and structure of mouse keratin intermediate filaments

    SciTech Connect

    Mack, J.W.; Torchia, D.A.; Steinert, P.M.

    1988-07-26

    The molecular dynamics and structural organization of mouse epidermal keratin intermediate filaments (IF) have been studied via solid-state nuclear magnetic resonance (NMR) experiments performed on IF labeled both in vivo and in vitro with isotopically enriched amino acids. As a probe of the organization of the peripheral glycine-rich end domains of the IF, carbon-13 NMR experiments have been performed on subfilamentous forms (prekeratin) and on IF reassembled in vitro that had been labeled with either (1-/sup 13/C)glycine or (2-/sup 13/C)glycine, as more than 90% of the glycines of the keratins are located in the end domains. Measurements of carbon relaxation times, nuclear Overhauser enhancements, and signal intensities show that the motions of the peptide backbone in the end domains are effectively isotropic. These results indicate that the end domains of IF are remarkably flexible and have little or no structural order. To probe the structural organization of the coiled-coil rod domains of the IF, separate samples of native keratin IF, raised in primary tissue culture, were labeled with L-(1-/sup 13/C)leucine, L-(/sup 2/H/sub 10/)leucine, or L-(2,3,3-/sup 2/H/sub 3/)leucine, as greater than 90% of the leucyl residues of the keratin IF types studied are located in the coiled coils which form the central core of IF. Deuterium NMR experiments performed on IF labeled with deuteriated leucines indeed reveal a marked degree of peptide backbone rigidity within the coiled coils, confirming the initial conclusions of the carbon-13 data. These data, demonstrating relative peptide backbone rigidity yet side-chain flexibility, are interpreted to mean that the coiled coils of these keratin IF are not tightly packed together but rather form a somewhat looser structure which permits a significant degree of side-chain mobility.

  6. Strategies for solid-state NMR in high-field Bitter and hybrid magnets

    NASA Astrophysics Data System (ADS)

    van Bentum, P. J. M.; Maan, J. C.; van Os, J. W. M.; Kentgens, A. P. M.

    2003-07-01

    Strategies for successful solid-state NMR experiments in high-field Bitter and hybrid magnets are discussed and implemented on an older generation 24 T system. In general, the stability and homogeneity of this type of magnets do not meet the requirements for most NMR experiments. Nevertheless, we will show that a new type of shimming with ferromagnetic inserts, combined with feedback control of the power-supply and NMR reference deconvolution can be used to obtain well-resolved solid-state NMR spectra. We will demonstrate that this approach is especially useful for the study of half-integer quadrupolar nuclei. Experimental results include static and Magic Angle Spinning (MAS) 27Al NMR spectra of the minerals sillimanite and andalusite.

  7. Is solid-state NMR enhanced by dynamic nuclear polarization?

    PubMed

    Lee, Daniel; Hediger, Sabine; De Paëpe, Gaël

    2015-01-01

    The recent trend of high-field (~5-20 T), low-temperature (~100 K) ssNMR combined with dynamic nuclear polarization (DNP) under magic angle spinning (MAS) conditions is analyzed. A brief overview of the current theory of hyperpolarization for so-called MAS-DNP experiments is given, along with various reasons why the DNP-enhancement, the ratio of the NMR signal intensities obtained in the presence and absence of microwave irradiation suitable for hyperpolarization, should not be used alone to gauge the value of performing MAS-DNP experiments relative to conventional ssNMR. This is demonstrated through a dissection of the current conditions required for MAS-DNP with particular attention to resulting absolute sensitivities and spectral resolution. Consequently, sample preparation methods specifically avoiding the surplus of glass-forming solvents so as to improve the absolute sensitivity and resolution are discussed, as are samples that are intrinsically pertinent for MAS-DNP studies (high surface area, amorphous, and porous). Owing to their pertinence, examples of recent applications on these types of samples where chemically-relevant information has been obtained that would have been impossible without the sensitivity increases bestowed by MAS-DNP are also detailed. Additionally, a promising further implementation for MAS-DNP is exampled, whereby the sensitivity improvements shown for (correlation) spectroscopy of nuclei at low natural isotopic abundance, facilitate internuclear distance measurements, especially for long distances (absence of dipolar truncation). Finally, we give some speculative perspectives for MAS-DNP. PMID:25779337

  8. Solid-state 17O NMR study of benzoic acid adsorption on metal oxide surfaces.

    PubMed

    Hagaman, Edward W; Chen, Banghao; Jiao, Jian; Parsons, William

    2012-02-01

    Solid-state (17)O NMR spectra of (17)O-labeled benzoic and anisic acids are reported and benzoic acid is used to probe the surface of metal oxides. Complexes formed when benzoic acid is dry mixed with mesoporous silica, and nonporous titania and alumina are characterized. Chemical reactions with silica are not observed. The nature of benzoic acid on silica is a function of the water content of the oxide. The acid disperses in the pores of the silica if the silica is in equilibrium with ambient laboratory humidity. The acid displays high mobility as evidenced by a liquid-like, Lorentzian resonance. Excess benzoic acid remains as the crystalline hydrogen-bonded dimer. Benzoic acid reacts with titania and alumina surfaces in equilibrium with laboratory air to form the corresponding titanium and aluminum benzoates. In both materials the oxygen of the (17)O-labeled acid is bound to the metal, showing the reaction proceeds by bond formation between oxygen deficient metal sites and the oxygen of the carboxylic acid. (27)Al MAS NMR confirms this mechanism for the reaction on alumina. Dry mixing of benzoic acid with alumina rapidly quenches pentacoordinate aluminum sites, excellent evidence that these sites are confined to the surface of the alumina particles. PMID:22245610

  9. Zinc Solid-State NMR Spectroscopy of Human Carbonic Anhydrase: Implications for the Enzymatic Mechanism.

    SciTech Connect

    Lipton, Andrew S.; Heck, Robert W.; Ellis, Paul D.

    2004-04-14

    Many zinc enzymes utilize zinc bound water as a critical component of a catalytic reaction. The Zn2+ ion activates water through ionization, polarization, or simple displacement depending upon the mechanistic details. The fate of one proton from the bound water is determined primarily by the influence of directly bound Zn-ligands, as well as hydrogen bonding with a secondary coordination sphere of side chains and/or bound waters within the protein. We have employed low temperature solid-state 67Zn NMR spectroscopy to probe the nature of the bonding at Zn2+ in human carbonic anhydrase isozyme II (CAII). In particular we wanted to characterize the 67Zn NMR parameters of the metal with both water and hydroxide as the fourth ligand, but instead we show that hydroxide is bound to Zn2+ over the pH range of 5 to 8.5. These results suggest the accepted mechanism of action of CAII needs to be revised. These data serve to provide further understanding of the observed pH dependence of the activity of this well studied protein.

  10. Structural Dynamics and Conformational Equilibria of SERCA Regulatory Proteins in Membranes by Solid-State NMR Restrained Simulations

    PubMed Central

    De Simone, Alfonso; Mote, Kaustubh R.; Veglia, Gianluigi

    2014-01-01

    Solid-state NMR spectroscopy is emerging as a powerful approach to determine structure, topology, and conformational dynamics of membrane proteins at the atomic level. Conformational dynamics are often inferred and quantified from the motional averaging of the NMR parameters. However, the nature of these motions is difficult to envision based only on spectroscopic data. Here, we utilized restrained molecular dynamics simulations to probe the structural dynamics, topology and conformational transitions of regulatory membrane proteins of the calcium ATPase SERCA, namely sarcolipin and phospholamban, in explicit lipid bilayers. Specifically, we employed oriented solid-state NMR data, such as dipolar couplings and chemical shift anisotropy measured in lipid bicelles, to refine the conformational ensemble of these proteins in lipid membranes. The samplings accurately reproduced the orientations of transmembrane helices and showed a significant degree of convergence with all of the NMR parameters. Unlike the unrestrained simulations, the resulting sarcolipin structures are in agreement with distances and angles for hydrogen bonds in ideal helices. In the case of phospholamban, the restrained ensemble sampled the conformational interconversion between T (helical) and R (unfolded) states for the cytoplasmic region that could not be observed using standard structural refinements with the same experimental data set. This study underscores the importance of implementing NMR data in molecular dynamics protocols to better describe the conformational landscapes of membrane proteins embedded in realistic lipid membranes. PMID:24940774

  11. Solid State OXYGEN-17 NMR Studies of Hydrate in Biomolecules and Deuterium NMR Studies of Chain Dynamics in Crystalline bis

    NASA Astrophysics Data System (ADS)

    Zhang, Huiming

    1993-01-01

    There are two parts in this dissertation. In part one (chapter II-V), solid state ^{17 }O (I = 5/2) NMR is established as a useful and complementary approach to ^2H NMR for studying dynamic disorder of water of hydration in biomolecules. A highly efficient probe is designed and constructed from available small components for conducting these experiments. This reliable probe generates intense RF fields with modest RF power and has a good signal to noise ratio (chapter II). Since ^{17 }O has a large quadrupolar coupling constant (Qcc ~ 7 MHz), a density operator formalism including the 2^{rm nd}-order quadrupolar Hamiltonian is developed for analysing excitation of ^{17} O solid state NMR signals for the +1/2 to -1/2 transition. A simple phase cycling echo sequence is developed for reducing the distortion caused by non-uniform excitation and the resultant patterns are compared with experimental spectra (chapter III). To analyze the lineshape of the central transition determined by the motionally averaged 2^{rm nd}-order quadrupolar perturbation, a 4 ^{rm th}-rank order parameter formalism for the fast motion limit (tau _{rm c} < 10^{-5} s) is developed. It is based on angular momentum coupling theory and requires a maximum of 31 order parameters. By a principal axis transformation, this number can be reduced to 8. To study slower motion, we established an adiabatic intermediate exchange theory for tau_ {rm c} ~ 10^{-5} s. The powder patterns calculated from these theories show good agreement when compared to the experimental spectra of several hydrates which undergo simple dynamics (chapter VI-V). In part two (chapter VI), solid state ^2H NMR is applied to explore phase transition related to chain dynamics in (Cd(CH_3CH _2CH_2CH _2NH_3)_2) Cl_4. This layered structure is a crystalline model of a lipid bilayer and was studied from 120K to 350K. Full order tensors for all sites labeled except for C^{(3)} were systematically measured. We find that changes in the chain dynamics are associated with the occurrence of phase transitions. For T < 120K, we find that the chain has an approximately all trans configuration with small "vibration" increasing in amplitude from the polar "head" of the chain to its hydrophilic "tail". For T > 200K, rapid, large amplitude dynamics of the chain vector caused by gauche-trans isomerization leads to significant tilting of the average chain direction. The order tensors are nonaxial showing the absence of axial rotation which is seen in the rotator phase of paraffins and the liquid crystal phase of lipids. In the range of 120K-200K, the symmetry of the dynamics decreases but substantial population of gauche isomers persists. However, the isomerization rate is slow on the ^2H NMR time scale of 10^{-5} s and spectral narrowing is due largely to small amplitude vibrations. The order tensors calculated from x-ray data or the usual "fixed" lattice rotational-isomeric model differ distinctly from the experimental tensors. We propose a flexible lattice model which successfully relates measured order tensors to the chain dynamics.

  12. Genetic algorithms and solid state NMR pulse sequences.

    PubMed

    Bechmann, Matthias; Clark, John; Sebald, Angelika

    2013-03-01

    The use of genetic algorithms for the optimisation of magic angle spinning NMR pulse sequences is discussed. The discussion uses as an example the optimisation of the C7(2)(1) dipolar recoupling pulse sequence, aiming to achieve improved efficiency for spin systems characterised by large chemical shielding anisotropies and/or small dipolar coupling interactions. The optimised pulse sequence is found to be robust over a wide range of parameters, requires only minimal a priori knowledge of the spin system for experimental implementations with buildup rates being solely determined by the magnitude of the dipolar coupling interaction, but is found to be less broadbanded than the original C7(2)(1) pulse sequence. The optimised pulse sequence breaks the synchronicity between r.f. pulses and sample spinning. PMID:23357428

  13. Weak and Transient Protein Interactions Determined by Solid-State NMR.

    PubMed

    Dannatt, Hugh R W; Felletti, Michele; Jehle, Stefan; Wang, Yao; Emsley, Lyndon; Dixon, Nicholas E; Lesage, Anne; Pintacuda, Guido

    2016-06-01

    Despite their roles in controlling many cellular processes, weak and transient interactions between large structured macromolecules and disordered protein segments cannot currently be characterized at atomic resolution by X-ray crystallography or solution NMR. Solid-state NMR does not suffer from the molecular size limitations affecting solution NMR, and it can be applied to molecules in different aggregation states, including non-crystalline precipitates and sediments. A solid-state NMR approach based on high magnetic fields, fast magic-angle sample spinning, and deuteration provides chemical-shift and relaxation mapping that enabled the characterization of the structure and dynamics of the transient association between two regions in an 80 kDa protein assembly. This led to direct verification of a mechanism of regulation of E. coli DNA metabolism. PMID:27101578

  14. Solid-state /sup 13/C NMR and X-ray diffraction of dermatan sulfate

    SciTech Connect

    Winter, W.T.; Taylor, M.G.; Stevens, E.S.; Morris, E.R.; Rees, D.A.

    1986-05-29

    Dermatan sulfate in the solid state has been studied by /sup 13/C CP/MAS nmr and X-ray diffraction in order to establish the ring conformation of the L-iduronate moiety. The solid state nmr spectrum is similar to the solution spectrum obtained previously, indicating that a ring conformation at least approximating to /sup 1/C/sub 4/ predominates in the solid state. X-ray powder diffraction data from the same sample indicate the presence of the 8-fold helix form previously observed by fiber diffraction, and interpreted in terms of a /sup 4/C/sub 1/ ring form. A likely explanation of the results is that a distorted /sup 1/C/sub 4/ L-iduronate ring conformation, not considered in the initial X-ray analysis, may emerge to provide a satisfactory interpretation of all available physical-chemical data.

  15. Investigation of Moisture Interaction with Cellulose Using Solid-State NMR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in Gossypium barbadense (Pima) cotton were investigated by 1H and 13C solid-state NMR techniques. 1H spin diffusion results indicate that water is present in multiple layers within the cotton fiber, each layer bei...

  16. Chemical structure of soil organic matter in slickspots as investigated by advanced solid-state NMR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Slickspot soils are saline, and knowledge of their humic chemistry would contribute to our limited understanding how salinity affects soil C and N stocks. We characterized humic acids (HAs) from slickspot soils with solid-state 13C nuclear magnetic resonance (NMR). Expanding on previous use of cross...

  17. 1H and 13C Solid-state NMR of Gossypium barbadense (Pima) Cotton

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in G. barbadense (Pima) cotton were investigated by 1H and 13C solid-state NMR techniques. 1H spin diffusion results from a Goldman-Shen experiment indicate that the water is multilayered. 1H MAS experiments pro...

  18. 1H and 13C Solid-state NMR of G. barbadense (Pima) Cotton

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in G. barbadense (Pima) cotton were investigated with 1H and 13C solid-state NMR techniques. 1H spin diffusion results from a Goldman-Shen experiment indicate that the water is multilayered. 1H MAS experiment...

  19. Solid-state NMR and ESR studies of activated carbons produced from pecan shells

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Activated carbon from pecan shells has shown promise as an adsorbent in water treatment and sugar refining. However, the chemistry of the material is complex and not fully understood. We report here the application of solid state NMR and ESR to study the chemical structure, mobility, and pore volu...

  20. High-Resolution Solid-State NMR Spectroscopy: Characterization of Polymorphism in Cimetidine, a Pharmaceutical Compound

    ERIC Educational Resources Information Center

    Pacilio, Julia E.; Tokarski, John T.; Quiones, Rosalynn; Iuliucci, Robbie J.

    2014-01-01

    High-resolution solid-state NMR (SSNMR) spectroscopy has many advantages as a tool to characterize solid-phase material that finds applications in polymer chemistry, nanotechnology, materials science, biomolecular structure determination, and others, including the pharmaceutical industry. The technology associated with achieving high resolution

  1. Characterization of animal manure using advanced solid-state C-13 NMR spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Knowledge of chemical structure of animal manure is necessary for its effective utilization. However, characterization of animal manure is challenging since it is a complex mixture and partially soluble. Solid-state C-13 NMR (nuclear magnetic resonance) spectroscopy is regarded as the best tool to i...

  2. High-Resolution Solid-State NMR Spectroscopy: Characterization of Polymorphism in Cimetidine, a Pharmaceutical Compound

    ERIC Educational Resources Information Center

    Pacilio, Julia E.; Tokarski, John T.; Quiñones, Rosalynn; Iuliucci, Robbie J.

    2014-01-01

    High-resolution solid-state NMR (SSNMR) spectroscopy has many advantages as a tool to characterize solid-phase material that finds applications in polymer chemistry, nanotechnology, materials science, biomolecular structure determination, and others, including the pharmaceutical industry. The technology associated with achieving high resolution…

  3. ANALYSIS OF POTENTIALLY MOBILE PHOSPHORUS IN ARABLE SOILS USING SOLID STATE NMR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In many intensive agroecosystems continued inputs of phosphorus (P) over many years can significantly increase soil P concentrations and the risk of P losses to surface waters. This study used solid-state 31P nuclear magnetic resonance (NMR) spectroscopy (high power decoupled (HPDec)/ magic angle s...

  4. Quantitative solid state {sup 13}C NMR measurements on kerogens

    SciTech Connect

    Maroto-Valer, M.M.; Love, G.D.; Snape, C.E.

    1996-12-31

    The use of the well-established techniques of high power decoupling, magic angle spinning (MAS) and cross-polarisation (CP) are used routinely to obtain high resolution {sup 13}C spectra of kerogens. However, due to unfortunate spin dynamics in CP, not all the carbons are observed. Results on a selection of type I sand II kerogens will be presented to demonstrate that the best strategy to obtain quantitative {sup 13}C NMR results is offered by a combination of a low magnetic field strength to minimize problems with spinning sidebands and the simple, albeit insensitive, Bloch decay or single pulse excitation (SPE) technique. Virtually all of the carbon in the type I and II kerogens and a wide range of coals has been observed using the SPE technique with the aromaticity and non-protonated aromatic carbon concentrations being invariably higher than those derived by the more rapid, but quantitatively unreliable CP method. Further, an extremely good correlation has been obtained between the carbon aromaticities and atomic H/C ratios for the samples investigated. The methodology has also been used to estimate the long methylene chain contents of kerogens and the extent of aromatisation that occurs in normal pyrolysis and pyrolysis under high hydrogen pressure (hydropyrolysis).

  5. Gas vesicles across kingdoms: a comparative solid state NMR study

    PubMed Central

    Daviso, Eugenio; Belenky, Marina; Griffin, Robert G.; Herzfeld, Judith

    2013-01-01

    The buoyancy organelles of aquatic microorganisms have to meet stringent specifications: allowing gases to equilibrate freely across the proteinaceous shell, preventing the condensation of water vapor inside the hollow cavity, and resisting collapse under hydrostatic pressures that vary with column depth. These properties are provided by the 7–8 kDa gas vesicle protein A (GvpA), repeats of which form all but small, specialized portions of the shell. Magic angle spinning NMR is uniquely capable of providing high resolution information on the fold and assembly of GvpA. Here we compare results for the gas vesicles of the haloarchae Halobacterium salinarum with those obtained previously for the cyanobacterium Anabaena flos-aquae. The data suggest that the two organisms follow similar strategies for avoiding water condensation. On the other hand, in its relatively shallow habitat, H. salinarum is able to avoid collapse with a less costly GvpA fold than is adopted by A. flos-aquae. PMID:23920491

  6. Methionine bound to Pd/γ-Al2O3 catalysts studied by solid-state (13)C NMR.

    PubMed

    Johnson, Robert L; Schwartz, Thomas J; Dumesic, James A; Schmidt-Rohr, Klaus

    2015-11-01

    The chemisorption and breakdown of methionine (Met) adsorbed on Pd/γ-Al2O3 catalysts were investigated by solid-state NMR. (13)C-enriched Met (ca. 0.4mg) impregnated onto γ-Al2O3 or Pd/γ-Al2O3 gives NMR spectra with characteristic features of binding to γ-Al2O3, to Pd nanoparticles, and oxidative or reductive breakdown of Met. The SCH3 groups of Met showed characteristic changes in chemical shift on γ-Al2O3 (13ppm) vs. Pd (19ppm), providing strong evidence for preferential binding to Pd, while the NC carbon generates a small resonance at 96ppm assigned to a distinct nonprotonated species bound to O or Pd. Additionally, NMR shows that the SCH3 groups of Met are mobile on γ-Al2O3 but immobilized by binding to Pd particles; on small Pd particles (ca. 4nm), the NCH groups undergo large-amplitude motions. In a reducing environment, Met breaks down by C-S bond cleavage followed by formation of C2-C4 organic acids. The SCH3 signal shifts to 22ppm, which is likely the signature of the principal species responsible for strong catalyst inhibition. These experiments demonstrate that solid-state magic-angle spinning NMR of (13)C-enriched Met can be a sensitive probe to investigate catalyst surfaces and characterize catalyst inhibition both before reaction and postmortem. PMID:26422257

  7. Solid-State (17)O NMR studies of organic and biological molecules: Recent advances and future directions.

    PubMed

    Wu, Gang

    2016-02-01

    This Trends article highlights the recent advances published between 2012 and 2015 in solid-state (17)O NMR for organic and biological molecules. New developments in the following areas are described: (1) new oxygen-containing functional groups, (2) metal organic frameworks, (3) pharmaceuticals, (4) probing molecular motion in organic solids, (5) dynamic nuclear polarization, and (6) paramagnetic coordination compounds. For each of these areas, the author offers his personal views on important problems to be solved and possible future directions. PMID:26651417

  8. Characterization of covalent protein conjugates using solid-state sup 13 C NMR spectroscopy

    SciTech Connect

    Garbow, J.R.; Fujiwara, Hideji; Sharp, C.R.; Logusch, E.W. )

    1991-07-23

    Cross-polarization magic-angle spinning (CPMAS) {sup 13}C NMR spectroscopy has been used to characterize covalent conjugates of alachlor, an {alpha}-chloroacetamide hapten, with glutathione (GSH) and bovine serum albumin (BSA). The solid-state NMR method demonstrates definitively the covalent nature of these conjugates and can also be used to characterize the sites of hapten attachment to proteins. Three different sites of alachlor binding are observed in the BSA system. Accurate quantitation of the amount of hapten covalently bound to GSH and BSA is reported. The solid-state {sup 13}C NMR technique can easily be generalized to study other small molecule/protein conjugates and can be used to assist the development and refinement of synthetic methods needed for the successful formation of such protein alkylation products.

  9. Solid-state 19F-NMR analysis of 19F-labeled tryptophan in gramicidin A in oriented membranes.

    PubMed Central

    Grage, Stephan L; Wang, Junfeng; Cross, Timothy A; Ulrich, Anne S

    2002-01-01

    The response of membrane-associated peptides toward the lipid environment or other binding partners can be monitored by solid-state NMR of suitably labeled side chains. Tryptophan is a prominent amino acid in transmembrane helices, and its (19)F-labeled analogues are generally biocompatible and cause little structural perturbation. Hence, we use 5F-Trp as a highly sensitive NMR probe to monitor the conformation and dynamics of the indole ring. To establish this (19)F-NMR strategy, gramicidin A was labeled with 5F-Trp in position 13 or 15, whose chi(1)/chi(2) torsion angles are known from previous (2)H-NMR studies. First, the alignment of the (19)F chemical shift anisotropy tensor within the membrane was deduced by lineshape analysis of oriented samples. Next, the three principal axes of the (19)F chemical shift anisotropy tensor were assigned within the molecular frame of the indole ring. Finally, determination of chi(1)/chi(2) for 5F-Trp in the lipid gel phase showed that the side chain alignment differs by up to 20 degrees from its known conformation in the liquid crystalline state. The sensitivity gain of (19)F-NMR and the reduction in the amount of material was at least 10-fold compared with previous (2)H-NMR studies on the same system and 100-fold compared with (15)N-NMR. PMID:12496101

  10. Solid-state NMR studies of theophylline co-crystals with dicarboxylic acids.

    PubMed

    Pindelska, Edyta; Sokal, Agnieszka; Szeleszczuk, Lukasz; Pisklak, Dariusz Maciej; Kolodziejski, Waclaw

    2014-11-01

    In this work, three polycrystalline materials containing co-crystals of theophylline with malonic, maleic, and glutaric acids were studied using (13)C, (15)N and (1)H solid-state NMR and FT-IR spectroscopy. The NMR assignments were supported by gauge including projector augmented waves (GIPAW) calculations of chemical shielding, performed using X-ray determined geometry. The experimental (13)C cross polarization/magic angle spinning (CP/MAS) NMR results and the calculated isotropic chemical shifts were in excellent agreement. A rapid and convenient method for theophylline co-crystals crystal structure analysis has been proposed for co-crystals, which are potentially new APIs. PMID:25194346

  11. Solid-State NMR Characterization of Mixed Phosphonic Acid Ligand Binding and Organization on Silica Nanoparticles.

    PubMed

    Davidowski, Stephen K; Holland, Gregory P

    2016-04-01

    As ligand functionalization of nanomaterials becomes more complex, methods to characterize the organization of multiple ligands on surfaces is required. In an effort to further the understanding of ligand-surface interactions, a combination of multinuclear ((1)H, (29)Si, (31)P) and multidimensional solid-state nuclear magnetic resonance (NMR) techniques was utilized to characterize the phosphonic acid functionalization of fumed silica nanoparticles using methylphosphonic acid (MPA) and phenylphosphonic acid (PPA). (1)H → (29)Si cross-polarization (CP)-magic angle spinning (MAS) solid-state NMR was used to selectively detect silicon atoms near hydrogen atoms (primarily surface species); these results indicate that geminal silanols are preferentially depleted during the functionalization with phosphonic acids. (1)H → (31)P CP-MAS solid-state NMR measurements on the functionalized silica nanoparticles show three distinct resonances shifted upfield (lower ppm) and broadened compared to the resonances of the crystalline ligands. Quantitative (31)P MAS solid-state NMR measurements indicate that ligands favor a monodentate binding mode. When fumed silica nanoparticles were functionalized with an equal molar ratio of MPA and PPA, the MPA bound the nanoparticle surface preferentially. Cross-peaks apparent in the 2D (1)H exchange spectroscopy (EXSY) NMR measurements of the multiligand sample at short mixing times indicate that the MPA and PPA are spatially close (≤5 Å) on the surface of the nanostructure. Furthermore, (1)H-(1)H double quantum-single quantum (DQ-SQ) back-to-back (BABA) 2D NMR spectra further confirmed that MPA and PPA are strongly dipolar coupled with observation of DQ intermolecular contacts between the ligands. DQ experimental buildup curves and simulations indicate that the average distance between MPA and PPA is no further than 4.2 ± 0.2 Å. PMID:26914738

  12. Analysis of RF heating and sample stability in aligned static solid-state NMR spectroscopy.

    PubMed

    Li, Conggang; Mo, Yiming; Hu, Jun; Chekmenev, Eduard; Tian, Changlin; Gao, Fei Philip; Fu, Riqiang; Gor'kov, Peter; Brey, William; Cross, Timothy A

    2006-05-01

    Sample instability during solid-state NMR experiments frequently arises due to RF heating in aligned samples of hydrated lipid bilayers. A new, simple approach for estimating sample temperature is used to show that, at 9.4 T, sample heating depends mostly on (1)H decoupling power rather than on (15)N irradiation in PISEMA experiments. Such heating for different sample preparations, including lipid composition, salt concentration and hydration level was assessed and the hydration level was found to be the primary parameter correlated with sample heating. The contribution to RF heating from the dielectric loss appears to be dominant under our experimental conditions. The heat generated by a single scan was approximately calculated from the Q values of the probe, to be a 1.7 degrees C elevation per single pulse sequence iteration under typical sample conditions. The steady-state sample temperature during PISEMA experiments can be estimated based on the method presented here, which correlates the loss factor with the temperature rise induced by the RF heating of the sample. PMID:16483809

  13. Quantification of protein secondary structure by (13)C solid-state NMR.

    PubMed

    Andrade, Fabiana Diuk; Forato, Lucimara Aparecida; Bernardes Filho, Rubens; Colnago, Luiz Alberto

    2016-05-01

    High-resolution (13)C solid-state NMR stands out as one of the most promising techniques to solve the structure of insoluble proteins featuring biological and technological importance. The simplest nuclear magnetic resonance (NMR) spectroscopy method to quantify the secondary structure of proteins uses the areas of carbonyl and alpha carbon peaks. The quantification obtained by fitting procedures depends on the assignment of the peaks to the structure, type of line shape, number of peaks to be used, and other parameters that are set by the operator. In this paper, we demonstrate that the analysis of (13)C NMR spectra by a pattern recognition method-based on the singular value decomposition (SVD) regression, which does not depend on the operator-shows higher correlation coefficients for α-helix and β-sheet (0.96 and 0.91, respectively) than Fourier transform infrared spectroscopy (FTIR) method. Therefore, the use of (13)C solid-state NMR spectra and SVD is a simple and reliable method for quantifying the secondary structures of insoluble proteins in solid-state. PMID:27068694

  14. Membrane structure and dynamics as viewed by solid-state NMR spectroscopy.

    PubMed

    Auger, M

    1997-10-01

    The purpose of the present study is the investigation of the structure and dynamics of biological membranes using solid-state nuclear magnetic resonance (NMR) spectroscopy. Two approaches are used in our laboratory. The first involves the measurement of high-resolution 13C and 1H spectra obtained by the magic angle spinning (MAS) technique while the second approach involves the measurement of 31P and 2H powder spectra in static samples. This paper will present some recent results obtained by high-resolution solid-state 1H NMR on the conformation of gramicidin A incorporated in a phosphatidylcholine bilayers. More specifically, we were able to observe changes in the gramicidin spectra as a function of the cosolubilization solvent initially used to prepare the samples. The interaction between lipid bilayers and an anticancer drug derived from chloroethylurea was also investigated using proton NMR spectroscopy. Finally, we have studied the interaction between cardiotoxin, a toxic protein extracted from snake venom, and negatively charged lipid bilayers using 31P solid-state NMR spectroscopy. PMID:9468622

  15. Magic-angle-spinning solid-state NMR of membrane proteins.

    PubMed

    Baker, Lindsay A; Folkers, Gert E; Sinnige, Tessa; Houben, Klaartje; Kaplan, Mohammed; van der Cruijsen, Elwin A W; Baldus, Marc

    2015-01-01

    Solid-state NMR spectroscopy (ssNMR) provides increasing possibilities to examine membrane proteins in different molecular settings, ranging from synthetic bilayers to whole cells. This flexibility often enables ssNMR experiments to be directly correlated with membrane protein function. In this contribution, we discuss experimental aspects of such studies starting with protein expression and labeling, leading to membrane protein isolation or to membrane proteins in a cellular environment. We show that optimized procedures can depend on aspects such as the achieved levels of expression, the stability of the protein during purification or proper refolding. Dealing with native membrane samples, such as isolated cellular membranes, can alleviate or entirely remove such biochemical challenges. Subsequently, we outline ssNMR experiments that involve the use of magic-angle-spinning and can be used to study membrane protein structure and their functional aspects. We pay specific attention to spectroscopic issues such as sensitivity and spectral resolution. The latter aspect can be controlled using a combination of tailored preparation procedures with solid-state NMR experiments that simplify the spectral analysis using specific filtering and correlation methods. Such approaches have already provided access to obtain structural views of membrane proteins and study their function in lipid bilayers. Ongoing developments in sample preparation and NMR methodology, in particular in using hyperpolarization or proton-detection schemes, offer additional opportunities to study membrane proteins close to their cellular function. These considerations suggest a further increase in the potential of using solid-state NMR in the context of prokaryotic or eukaryotic membrane protein systems in the near future. PMID:25950971

  16. Ab Initio Quality NMR Parameters in Solid-State Materials Using a High-Dimensional Neural-Network Representation.

    PubMed

    Cuny, Jérôme; Xie, Yu; Pickard, Chris J; Hassanali, Ali A

    2016-02-01

    Nuclear magnetic resonance (NMR) spectroscopy is one of the most powerful experimental tools to probe the local atomic order of a wide range of solid-state compounds. However, due to the complexity of the related spectra, in particular for amorphous materials, their interpretation in terms of structural information is often challenging. These difficulties can be overcome by combining molecular dynamics simulations to generate realistic structural models with an ab initio evaluation of the corresponding chemical shift and quadrupolar coupling tensors. However, due to computational constraints, this approach is limited to relatively small system sizes which, for amorphous materials, prevents an adequate statistical sampling of the distribution of the local environments that is required to quantitatively describe the system. In this work, we present an approach to efficiently and accurately predict the NMR parameters of very large systems. This is achieved by using a high-dimensional neural-network representation of NMR parameters that are calculated using an ab initio formalism. To illustrate the potential of this approach, we applied this neural-network NMR (NN-NMR) method on the (17)O and (29)Si quadrupolar coupling and chemical shift parameters of various crystalline silica polymorphs and silica glasses. This approach is, in principal, general and has the potential to be applied to predict the NMR properties of various materials. PMID:26730889

  17. Solid-state NMR approaches to measure topological equilibria and dynamics of membrane polypeptides.

    PubMed

    Salnikov, Evgeniy; Aisenbrey, Christopher; Vidovic, Verica; Bechinger, Burkhard

    2010-02-01

    Biological membranes are characterized by a high degree of dynamics. In order to understand the function of membrane proteins and even more of membrane-associated peptides, these motional aspects have to be taken into consideration. Solid-state NMR spectroscopy is a method of choice when characterizing topological equilibria, molecular motions, lateral and rotational diffusion as well as dynamic oligomerization equilibria within fluid phase lipid bilayers. Here we show and review examples where the (15)N chemical shift anisotropy, dipolar interactions and the deuterium quadrupolar splittings have been used to analyze motions of peptides such as peptaibols, antimicrobial sequences, Vpu, phospholamban or other channel domains. In particular, simulations of (15)N and (2)H-solid-state NMR spectra are shown of helical domains in uniaxially oriented membranes when rotation around the membrane normal or the helix long axis occurs. PMID:19596252

  18. Direct observation of zeolite a synthesis by in situ solid-state NMR

    SciTech Connect

    Shi, J.; Anderson, M.W.; Carr, S.W.

    1996-02-01

    This paper describes the use of in situ solid-state NMR and X-ray powder diffraction to study the real-time synthesis of zeolite. In particular {sup 27}Al and {sup 29}Si are used to monitor the growth in situ X-ray diffraction study was used to investigate the development of long range order of the material. Conclusions concerning the mechanism of the formation of zeolite A are proposed. 28 refs., 12 figs., 2 tabs.

  19. Benchmarks for the 13C NMR chemical shielding tensors in peptides in the solid state

    NASA Astrophysics Data System (ADS)

    Czernek, Jiří; Pawlak, Tomasz; Potrzebowski, Marek J.

    2012-02-01

    The benchmark set is proposed, which comprises 126 principal elements of chemical shielding tensors, and the respective isotropic chemical shielding values, of all 42 13C nuclei in crystalline Tyr-D-Ala-Phe and Tyr-Ala-Phe tripeptides with known, but highly dissimilar structures. These data are obtained by both the NMR measurements and the density functional theory in the pseudopotential plane-wave scheme. Using the CASTEP program, several computational strategies are employed, for which the level of agreement between calculations and experiment is established. This set is mainly intended for the validation of methods capable of predicting the 13C NMR parameters in solid-state systems.

  20. Detergent Optimized Membrane Protein Reconstitution in Liposomes for Solid State NMR

    PubMed Central

    2015-01-01

    For small helical membrane proteins, their structures are highly sensitive to their environment, and solid state NMR is a structural technique that can characterize these membrane proteins in native-like lipid bilayers and proteoliposomes. To date, a systematic method by which to evaluate the effect of the solubilizing detergent on proteoliposome preparations for solid state NMR of membrane proteins has not been presented in the literature. A set of experiments are presented aimed at determining the conditions most amenable to dialysis mediated reconstitution sample preparation. A membrane protein from M. tuberculosis is used to illustrate the method. The results show that a detergent that stabilizes the most protein is not always ideal and sometimes cannot be removed by dialysis. By focusing on the lipid and protein binding properties of the detergent, proteoliposome preparations can be readily produced, which provide double the signal-to-noise ratios for both the oriented sample and magic angle spinning solid state NMR. The method will allow more membrane protein drug targets to be structurally characterized in lipid bilayer environments. PMID:24665863

  1. LARGE SCALE PRODUCTION, PURIFICATION, AND 65CU SOLID STATE NMR OF AZURIN

    SciTech Connect

    Gao, A.; Heck, R.W.

    2008-01-01

    This paper details a way to produce azurin with an effi ciency over 10 times greater than previously described and demonstrates the fi rst solid state nuclear magnetic resonance spectrum of 65Cu(I) in a metalloprotein. A synthetic gene for azurin based upon the DNA sequence from Pseudomonas aeruginosa including the periplasmic targeting sequence was subcloned into a T7 overexpression vector to create the plasmid pGS-azurin, which was transformed into BL21 (DE3) competent cells. The leader sequence on the expressed protein causes it to be exported to the periplasmic space of Escherichia coli. Bacteria grown in a fermentation unit were induced to overexpress the azurin, which was subsequently purifi ed through an endosmotic shock procedure followed by high performance liquid chromatography (HPLC). 1,500 mg of azurin were purifi ed per liter of culture. 65Cu(II) was added to apo-azurin and then reduced. The 65Cu metal cofactor in azurin was observed with solid state nuclear magnetic resonance (NMR) to determine any structural variations that accompanied copper reduction. This is the fi rst solid state NMR spectra of a copper(I) metalloprotein. Analysis of the NMR spectra is being used to complement hypotheses set forth by x-ray diffraction and computational calculations of electron transfer mechanisms in azurin.

  2. Mechanism of dilute-spin-exchange in solid-state NMR

    SciTech Connect

    Lu, George J.; Opella, Stanley J.

    2014-03-28

    In the stationary, aligned samples used in oriented sample (OS) solid-state NMR, {sup 1}H-{sup 1}H homonuclear dipolar couplings are not attenuated as they are in magic angle spinning solid-state NMR; consequently, they are available for participation in dipolar coupling-based spin-exchange processes. Here we describe analytically the pathways of {sup 15}N-{sup 15}N spin-exchange mediated by {sup 1}H-{sup 1}H homonuclear dipolar couplings. The mixed-order proton-relay mechanism can be differentiated from the third spin assisted recoupling mechanism by setting the {sup 1}H to an off-resonance frequency so that it is at the “magic angle” during the spin-exchange interval in the experiment, since the “magic angle” irradiation nearly quenches the former but only slightly attenuates the latter. Experimental spectra from a single crystal of N-acetyl leucine confirm that this proton-relay mechanism plays the dominant role in {sup 15}N-{sup 15}N dilute-spin-exchange in OS solid-state NMR in crystalline samples. Remarkably, the “forbidden” spin-exchange condition under “magic angle” irradiation results in {sup 15}N-{sup 15}N cross-peaks intensities that are comparable to those observed with on-resonance irradiation in applications to proteins. The mechanism of the proton relay in dilute-spin-exchange is crucial for the design of polarization transfer experiments.

  3. Monitoring the Electrochemical Processes in the Lithium–Air Battery by Solid State NMR Spectroscopy

    PubMed Central

    2013-01-01

    A multi-nuclear solid-state NMR approach is employed to investigate the lithium–air battery, to monitor the evolution of the electrochemical products formed during cycling, and to gain insight into processes affecting capacity fading. While lithium peroxide is identified by 17O solid state NMR (ssNMR) as the predominant product in the first discharge in 1,2-dimethoxyethane (DME) based electrolytes, it reacts with the carbon cathode surface to form carbonate during the charging process. 13C ssNMR provides evidence for carbonate formation on the surface of the carbon cathode, the carbonate being removed at high charging voltages in the first cycle, but accumulating in later cycles. Small amounts of lithium hydroxide and formate are also detected in discharged cathodes and while the hydroxide formation is reversible, the formate persists and accumulates in the cathode upon further cycling. The results indicate that the rechargeability of the battery is limited by both the electrolyte and the carbon cathode stability. The utility of ssNMR spectroscopy in directly detecting product formation and decomposition within the battery is demonstrated, a necessary step in the assessment of new electrolytes, catalysts, and cathode materials for the development of a viable lithium–oxygen battery. PMID:24489976

  4. Solid-state NMR: An emerging technique in structural biology of self-assemblies.

    PubMed

    Habenstein, Birgit; Loquet, Antoine

    2016-03-01

    Protein self-assemblies are ubiquitous biological systems involved in many cellular processes, ranging from bacterial and viral infection to the propagation of neurodegenerative disorders. Studying the atomic three-dimensional structures of protein self-assemblies is a particularly demanding task, as these systems are usually insoluble, non-crystalline and of large size. Solid-state NMR (ssNMR) is an emerging method that can provide atomic-level structural data on intact macromolecular assemblies. We here present recent progress in magic-angle spinning ssNMR to study protein assemblies and give an overview on its combination with complementary techniques such as cryo-EM, mass-per-length measurements, SAXS and X-ray diffraction. Applications of ssNMR on its own and in hybrid approaches have revealed precious atomic details and first high-resolution structures of complex biological assemblies, including amyloid fibrils, bacterial filaments, phages or virus capsids. PMID:26234527

  5. High-resolution solid-state NMR studies on uniformly [13C,15N]-labeled ubiquitin.

    PubMed

    Seidel, Karsten; Etzkorn, Manuel; Heise, Henrike; Becker, Stefan; Baldus, Marc

    2005-09-01

    Understanding of the effects of intermolecular interactions, molecular dynamics, and sample preparation on high-resolution magic-angle spinning NMR data is currently limited. Using the example of a uniformly [13C,15N]-labeled sample of ubiquitin, we discuss solid-state NMR methods tailored to the construction of 3D molecular structure and study the influence of solid-phase protein preparation on solid-state NMR spectra. A comparative analysis of 13C', 13Calpha, and 13Cbeta resonance frequencies suggests that 13C chemical-shift variations are most likely to occur in protein regions that exhibit an enhanced degree of molecular mobility. Our results can be refined by additional solid-state NMR techniques and serve as a reference for ongoing efforts to characterize the structure and dynamics of (membrane) proteins, protein complexes, and other biomolecules by high-resolution solid-state NMR. PMID:16094694

  6. Preparation of protein nanocrystals and their characterization by solid state NMR.

    PubMed

    Martin, Rachel W; Zilm, Kurt W

    2003-11-01

    Preparation of proteins in their crystalline state has been found to be important in producing stable therapeutic protein formulations, cross-linked enzyme crystals for application in industrial processes, generating novel porous media for separations, and of course in structure elucidation. Of these applications only X-ray crystallography requires large crystals, defined here as being crystals 100s of microns or greater in size. Smaller crystals have attractive attributes in many instances, and are just as useful in structure determination by solid state NMR (ssNMR) as are large crystals. In this paper we outline a simple set of procedures for preparing nanocrystalline protein samples for ssNMR or other applications and describe the characterization of their crystallinity by ssNMR and X-ray powder diffraction. The approach is demonstrated in application to five different proteins: ubiquitin, lysozyme, ribonuclease A, streptavidin, and cytochrome c. In all instances the nanocrystals produced are found to be highly crystalline as judged by natural abundance 13C ssNMR and optical and electron microscopy. We show for ubiquitin that nanocrystals prepared by rapid batch crystallization yield equivalent 13C ssNMR spectra to those of larger X-ray diffraction quality crystals. Single crystal and powder X-ray diffraction measurements are made to compare the degree of order present in polycrystalline, nanocrystalline, and lyophilized ubiquitin. Solid state 13C NMR is also used to show that ubiquitin nanocrystals are thermally robust, giving no indication of loss of local order after repeated temperature cycling between liquid nitrogen and room temperature. The methods developed are rapid and should scale well from the tenths of milligram to multi-gram scales, and as such should find wide utility in the preparation of protein nanocrystals for applications in catalysis, separations, and especially in sample preparation for structural studies using ssNMR. PMID:14568526

  7. Solid-State (63)Cu, (65)Cu, and (31)P NMR Spectroscopy of Photoluminescent Copper(I) Triazole Phosphine Complexes.

    PubMed

    Yu, Huaguang; Tan, Xiuzhen; Bernard, Guy M; Terskikh, Victor V; Chen, Jinglin; Wasylishen, Roderick E

    2015-07-30

    The results of a solid-state (63/65)Cu and (31)P NMR investigation of several copper(I) complexes with functionalized 3-(2'-pyridyl)-1,2,4-triazole and phosphine ligands that have shown potential in the preparation of photoluminescent devices are reported. For each complex studied, distinct NMR parameters, with moderate (63)Cu nuclear quadrupolar coupling constant (CQ) values ranging from -17.2 to -23.7 MHz, are attributed to subtle variations in the distorted four-coordinate environments about the copper nuclei. The spans of the copper chemical shift (CS) tensors, δ11-δ33, for the mono- and bisphosphine complexes are also similar, ranging from 1000 to 1150 ppm, but that for a complex with a strained bidentate phosphine ligand is only 650 ppm. The effects of residual dipolar and indirect spin-spin coupling arising from the (63/65)Cu- (31)P spin pairs, observed in the solid-state (31)P NMR spectra of these complexes, yield information about the orientations of the copper electric field gradient (EFG) tensors relative to the Cu-P bond. Variable-temperature (31)P NMR measurements for [Cu(bptzH)(dppe)]ClO4 (bptzH = 5-tert-butyl-3-(2'-pyridyl)-1,2,4-triazole; dppe = 1,2-bis(diphenylphosphino)ethane), undertaken to investigate the cause of the broad unresolved spectra observed at room temperature, demonstrate that the broadening arises from partial self-decoupling of the (63/65)Cu nuclei, a consequence of rapid quadrupolar relaxation. Ab initio calculations of copper EFG and CS tensors were performed to probe relationships between NMR parameters and molecular structure. The analysis demonstrated that CQ((63/65)Cu) is negative for all complexes studied here and that the largest components of the EFG tensors are generally coincident with δ11. PMID:26101890

  8. Solid State NMR Measurements for Preliminary Lifetime Assessments in (gamma)-Irradiated and Thermally Aged Siloxane Elastomers

    SciTech Connect

    Chinn, S C; Herberg, J L; Sawvel, A M; Maxwell, R S

    2005-02-03

    Siloxanes have a wide variety of applications throughout the aerospace industry which take advantage of their exceptional insulating and adhesive properties and general resilience. They also offer a wide range of tailorable engineering properties with changes in composition and filler content. They are, however, subject to degradation in radiatively and thermally harsh environments. We are using solid state nuclear magnetic resonance techniques to investigate changes in network and interfacial structure in siloxane elastomers and their correlations to changes in engineering performance in a series of degraded materials. Nuclear magnetic resonance (NMR) parameters such as transverse (T{sub 2}) relaxation times, cross relaxation rates, and residual dipolar coupling constants provide excellent probes of changes crosslink density and motional dynamics of the polymers caused by multi-mechanism degradation. The results of NMR studies on aged siloxanes are being used in conjunction with other mechanical tests to provide insight into component failure and degradation kinetics necessary for preliminary lifetime assessments of these materials as well as into the structure-property relationships of the polymers. NMR and magnetic resonance imaging (MRI) results obtained both from high resolution NMR spectrometers as well as low resolution benchtop NMR screening tools will be presented.

  9. Solid State NMR Measurements for Preliminary Lifetime Assessments in gamma-Irradiated and Thermally Aged Siloxane Elastomers

    SciTech Connect

    Chinn, S C; Herberg, J L; Sawvel, A M; Maxwell, R S

    2004-11-29

    Siloxanes have a wide variety of applications throughout the aerospace industry which take advantage of their exceptional insulating and adhesive properties and general resilience. They also offer a wide range of tailorable engineering properties with changes in composition and filler content. They are, however, subject to degradation in radiatively and thermally harsh environments. We are using solid state nuclear magnetic resonance techniques to investigate changes in network and interfacial structure in siloxane elastomers and their correlations to changes in engineering performance in a series of degraded materials. NMR parameters such as transverse ( T{sub 2}) relaxation times, cross relaxation rates, and residual dipolar coupling constants provide excellent probes of changes crosslink density and motional dynamics of the polymers caused by multi-mechanism degradation. The results of NMR studies on aged siloxanes are being used in conjunction with other mechanical tests to provide insight into component failure and degradation kinetics necessary for preliminary lifetime assessments of these materials as well as into the structure-property relationships of the polymers. NMR and MRI results obtained both from high resolution NMR spectrometers as well as low resolution benchtop NMR screening tools will be presented.

  10. Structure and dynamics of retinal in rhodopsin elucidated by deuterium solid state NMR

    NASA Astrophysics Data System (ADS)

    Salgado, Gilmar Fernandes De Jesus

    Rhodopsin is a seven transmembrane helix GPCR found which mediates dim light vision, in which the binding pocket is occupied by the ligand 11- cis-retinal. A site-directed 2H-labeling approach utilizing solid-state 2H NMR spectroscopy was used to investigate the structure and dynamics of retinal within its binding pocket in the dark state of rhodopsin, and as well the MetaI and MetaII. 11-cis-[5-C 2H3]-, 11-cis-[9-C 2H3]-, and 11-cis-[13-C2H 3]-retinal were used to regenerate bleached rhodopsin. Recombinant membranes comprising purified rhodopsin and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were prepared (1:50 molar ratio). Solid-state 2H NMR spectra were obtained for the aligned rhodopsin/POPC recombinant membranes at temperatures below the order-disorder phase transition temperature of POPC. The solid-state NMR studies of aligned samples, give the orientations of the 2H nuclear coupling tensor relative to the membrane frame, which involve both the conformation and orientation of the bound retinal chromophore. Theoretical simulations of the experimental 2H NMR spectra employed a new lineshape treatment for a semi-random distribution due to static uniaxial disorder. The analysis gives the orientation of the 2H-labeled C-C2H3 methyl bond axes relative to the membrane plane as well as the extent of three-dimensional alignment disorder (mosaic spread). These results clearly demonstrate the applicability of site-directed 2H NMR methods for investigating conformational changes and dynamics of ligands bound to rhodopsin and other GPCRs in relation to their characteristic mechanisms of action.

  11. 1H to 13C Energy Transfer in Solid State NMR Spectroscopy of Natural Organic Systems

    NASA Astrophysics Data System (ADS)

    Berns, Anne E.; Conte, Pellegrino

    2010-05-01

    Cross polarization (CP) magic angle spinning (MAS) 13C-NMR spectroscopy is a solid state NMR technique widely used to study chemical composition of organic materials with low or no solubility in the common deuterated solvents used to run liquid state NMR experiments. Based on the magnetization transfer from abundant nuclei (with spin of 1 -2) having a high gyromagnetic ratio (γ), such as protons, to the less abundant 13C nuclei with low γ values, 13C-CPMAS NMR spectroscopy is often applied in environmental chemistry to obtain quantitative information on the chemical composition of natural organic matter (NOM) (Conte et al., 2004), although its quantitative assessment is still matter of heavy debates. Many authors (Baldock et al., 1997; Conte et al., 1997, 2002; Dria et al., 2002; Kiem et al., 2000; Kögel-Knabner, 2000; Preston, 2001), reported that the application of appropriate instrument setup as well as the use of special pulse sequences and correct spectra elaboration may provide signal intensities that are directly proportional to the amount of nuclei creating a NMR signal. However, many other papers dealt with the quantitative unsuitability of 13C-CPMAS NMR spectroscopy. Among those, Mao et al. (2000), Smernik and Oades (2000 a,b), and Preston (2001) reported that cross-polarized NMR techniques may fail in a complete excitation of the 13C nuclei. In fact, the amount of observable carbons via 13C-CPMAS NMR spectroscopy appeared, in many cases, lower than that measured by a direct observation of the 13C nuclei. As a consequence, cross-polarized NMR techniques may provide spectra where signal distribution may not be representative of the quantitative distribution of the different natural organic matter components. Cross-polarization is obtained after application of an initial 90° x pulse on protons and a further spin lock pulse (along the y axis) having a fixed length (contact time) for both nuclei (1H and 13C) once the Hartmann-Hahn condition is matched. The Hartmann-Hahn condition can be expressed as γHB1H = γCB1C, where γH and γC are the gyromagnetic ratios of protons and carbons, whereas B1H and B1C are the 1H and 13C radio-frequency (r.f.) fields applied to the nuclei. The Hartmann-Hahn condition is affected by the H-C dipolar interaction strength (Stejskal & Memory, 1994). All the factors affecting dipolar interactions may mismatch the Hartmann-Hahn condition and prevent a quantitative representation of the NOM chemical composition (Conte et al., 2004). It has been reported that under low speed MAS conditions, broad matching profiles are centered around the Hartmann-Hahn condition....... With increasing spinning speed the Hartmann-Hahn matching profiles break down in a series of narrow matching bands separated by the rotor frequency (Stejskal & Memory, 1994). In order to account for the instability of the Hartmann-Hahn condition at higher rotor spin rates (>10 kHz), variable amplitude cross-polarization techniques (RAMP-CP) have been developed (Metz et al., 1996). So far, to our knowledge, the prevailing way used to obtain quantitative 13C-CPMAS NMR results was to optimize the 1H and 13C spin lock r.f. fields on simple standard systems such as glycine and to use those r.f. field values to run experiments on unknown organic samples. The aim of the present study was to experimentally evidence that the stability of the Hartmann-Hahn condition was different for different samples with a known structure. Moreover, Hartmann-Hahn profiles of four different humic acids (HAs) were also provided in order to show that the 1H/13C r.f. spin lock field strength must also be tested on the HAs prior to a quantitative evaluation of their 13C-CPMAS NMR spectra. Baldock, J.A., Oades, J.M., Nelson, P.N., Skene, T.M., Golchin, A. & Clarke, P., 1997. Assessing the extent of decomposition of natural organic materials using solid-state C-13 NMR spectroscopy. Australian Journal of Soil Research, 35, 1061-1083. Conte, P., Piccolo, A., van Lagen, B., Buurman, P. & de Jager, P.A., 1997. Quantitative Aspects of Solid-State 13C-NMR Spectra of Humic Substances from Soils of Volcanic Systems. Geoderma, 80, 327-338. Conte, P., Piccolo, A., van Lagen, B., Buurman, P. & Hemminga, M.A., 2002. Elemental quantitation of natural organic matter by CPMAS C-13 NMR spectroscopy. Solid State Nuclear Magnetic Resonance, 21, 158-170. Conte, P., Spaccini, R. & Piccolo, A., 2004. State of the art of CPMAS C-13-NMR spectroscopy applied to natural organic matter. Progress in Nuclear Magnetic Resonance Spectroscopy, 44, 215-223. Dria, K.J., Sachleben, J.R. & Hatcher, P.G., 2002. Solid-state carbon-13 nuclear magnetic resonance of humic acids at high magnetic field strengths. Journal of Environmental Quality, 31, 393-401. Kiem, R., Knicker, H., Korschens, M. & Kogel-Knabner, I., 2000. Refractory organic carbon in C-depleted arable soils, as studied by C-13 NMR spectroscopy and carbohydrate analysis. Organic Geochemistry, 31, 655-668. Kögel-Knabner, I., 2000. Analytical approaches for characterizing soil organic matter. Organic Geochemistry, 31, 609-625. Mao, J.D., Hu, W.G., Schmidt-Rohr, K., Davies, G., Ghabbour, E.A. & Xing, B., 2000. Quantitative characterization of humic substances by solid-state carbon-13 nuclear magnetic resonance. Soil Science Society of America Journal, 64, 873-884. Metz, G., Ziliox, M. & Smith, S.O., 1996. Towards quantitative CP-MAS NMR. Solid State Nuclear Magnetic Resonance, 7, 155-160. Preston, C.M., 2001. Carbon-13 solid-state NMR of soil organic matter - using the technique effectively. Canadian Journal of Soil Science, 81, 255-270. Smernik, R.J. & Oades, J.M., 2000a. The use of spin counting for determining quantitation in solid state C-13 NMR spectra of natural organic matter 1. Model systems and the effects of paramagnetic impurities. Geoderma, 96, 101-129. Smernik, R.J. & Oades, J.M., 2000b. The use of spin counting for determining quantitation in solid state C-13 NMR spectra of natural organic matter 2. HF-treated soil fractions. Geoderma, 96, 159-171. Stejskal, E.O. & Memory, J.D., 1994. High Resolution NMR in the Solid State. Fundamentals of CP/MAS. Oxford University Press, New York.

  12. Ceramide Lipid Interactions studied by MD Simulations and Solid-State NMR

    PubMed Central

    Dutagaci, Bercem; Becker-Baldus, Johanna; Faraldo-Gmez, Jos D.; Glaubitz, Clemens

    2014-01-01

    Ceramides play a key modulatory role in many cellular processes, which results from their effect on the structure and dynamics of biological membranes. In this study, we investigate the influence of C16-ceramide (C16) on the biophysical properties of DMPC lipid bilayers using solid-state NMR and atomistic molecular dynamics (MD) simulations. MD simulations and NMR measurements were carried out for a pure DMPC bilayer and for a 20% DMPC-C16 mixture. Calculated key structural properties, namely area per lipid, chain order parameters, and mass density profiles, indicate that C16 has an ordering effect on the DMPC bilayer. Furthermore, the simulations predict that specific hydrogen-bonds form between DMPC and C16 molecules. Multi-nuclear solid-state NMR was used to verify these theoretical predictions. Chain order parameters extracted from 13C-1H dipole couplings were measured for both lipid and ceramide and follow the trend suggested by the MD simulations. Furthermore, 1H-MAS NMR experiments showed a direct contact between ceramide and lipids. PMID:24882733

  13. Structural characterization of nylon 7 by solid-state NMR, DSC, and ATR-FTIR

    NASA Astrophysics Data System (ADS)

    Johnson, C. G.; Mathias, L. J.

    1993-06-01

    Samples of commercial nylon 7 were given different thermal or precipitation histories. Structure and crystallinity were followed by DSC, solid-state NMR, and attenuated total reflectance FTIR (ATR-FTIR). Delta H and T(sub m) values ranged from 52 to 93 J/g and 228 to 242 C, respectively. Surprisingly, annealing did not give material with the greatest delta H although it possessed the highest melting point. Solid-state C-13 and N-15 NMR methods were used to observe the amorphous and crystalline fractions. For example, the amide nitrogens show resonances near 86.5 ppm in amorphous domains, near 84 ppm in alpha-crystals, and near 89 ppm in gamma-crystals. N-15 CP/MAS spectra of solution cast samples contained peaks consistent with all three domains in various intensity ratios but with the gamma-peak being the most intense for most samples. Solid-state C-13 spectra contained peaks which supported the presence of these phases but with the alpha-phase peaks predominating.

  14. Freezing Point Depression of Water in Phospholipid Membranes — A Solid-State NMR Study

    PubMed Central

    Lee, Dong-Kuk; Kwon, ByungSoo; Ramamoorthy, Ayyalusamy

    2009-01-01

    Lipid-water interaction plays an important role in the properties of lipid bilayers, cryoprotectants, and membrane-associated peptides and proteins. The temperature at which water bound to lipid bilayers freezes is lower than that of free water. Here, we report a solid-state NMR investigation on the freezing point depression of water in phospholipid bilayers in the presence and absence of cholesterol. Deuterium NMR spectra at different temperatures ranging from -75 to +10°C were obtained from fully 2H2O-hydrated POPC (1-palmitoyl-2-oleoyl-phosphatidylcholine) multilamellar vesicles (MLVs), prepared with and without cholesterol, to determine the freezing temperature of water and the effect of cholesterol on the freezing temperature of water in POPC bilayers. Our 2H NMR experiments reveal the motional behavior of unfrozen water molecules in POPC bilayers even at temperatures significantly below 0°C and show that the presence of cholesterol further lowered the freezing temperature of water in POPC bilayers. These results suggest that in the presence of cholesterol the fluidity and dynamics of lipid bilayers can be retained even at very low temperatures as exist in the liquid crystalline phase of the lipid. Therefore, bilayer samples prepared with a cryoprotectant like cholesterol should enable the performance of multidimensional solid-state NMR experiments to investigate the structure, dynamics, and topology of membrane proteins at a very low temperature with enhanced sample stability and possibly a better sensitivity. Phosphorus-31 NMR data suggest that lipid bilayers can be aligned at low temperatures, while 15N NMR experiments demonstrate that such aligned samples can be used to enhance the signal-to-noise ratio of 15N chemical shift spectra of a 37-residue human antimicrobial peptide, LL-37. PMID:18991419

  15. Hydration-optimized oriented phospholipid bilayer samples for solid-state NMR structural studies of membrane proteins.

    PubMed

    Marassi, Francesca M; Crowell, Kevin J

    2003-03-01

    The preparation of oriented, hydration-optimized lipid bilayer samples, for NMR structure determination of membrane proteins, is described. The samples consist of planar phospholipid bilayers, containing membrane proteins, that are oriented on single pairs of glass slides, and are placed in the coil of the NMR probe with the bilayer plane perpendicular to the direction of the magnetic field. Lipid bilayers provide a medium that closely resembles the biological membrane, and sample orientation both preserves the intrinsic membrane-defined directional quality of membrane proteins, and provides the mechanism for resonance line narrowing. The hydration-optimized samples overcome some of the difficulties associated with multi-dimensional, high-resolution, solid-state NMR spectroscopy of membrane proteins. These samples have greater stability over the course of multi-dimensional NMR experiments, they have lower sample conductance for greater rf power efficiency, and enable greater rf coil filling factors to be obtained for improved experimental sensitivity. Sample preparation is illustrated for the membrane protein CHIF (channel inducing factor), a member of the FXYD family of ion transport regulators. PMID:12660112

  16. Interactions between Nafion resin and protonated dodecylamine modified montmorillonite: a solid state NMR study.

    PubMed

    Zhang, Limin; Xu, Jun; Hou, Guangjin; Tang, Huiru; Deng, Feng

    2007-07-01

    A series of nanocomposites have been prepared from perfluorosulfonylfluoride copolymer resin (Nafion) and layered montmorillonite (MMT) modified with protonated dodecylamine by conventional sol-gel intercalation. The structure of these nanocomposite materials have been characterized using FT-IR, elemental analysis, XRD and solid state NMR techniques, including 19F magic-angle spinning (MAS) NMR, 19F NMR relaxation time measurements, 29Si MAS, 1H MAS, 1H-13C cross-polarization magic-angle spinning (CPMAS), and 1H-13C heteronuclear correlation (HETCOR) 2D NMR. The results showed that thermal stability of Nafion was improved moderately by the addition of dodecylamine modified MMT without intercalation. FT-IR and 29Si MAS NMR results indicated that dodecylamine modification did not result in obvious changes in the MMT lattice structure. The XRD results showed that the protonated dodecylamine has been embedded and intercalated into the MMT interlayers, whereas Nafion was not. Elemental analysis results also suggested that some dodecylamine was adsorbed on the surface of MMT. 1H-13C HETCOR 2D NMR experiment clearly indicated that strong electrostatic interactions were present between the NH+3 group of dodecylamine and the fluorine-containing groups (CF3, OCF2, and SCF2) of Nafion resin. Such electrostatic interactions are probably the major contributors for the improved thermal stability of the resultant composite materials. PMID:17382953

  17. Characterization of Two Distinct Amorphous Forms of Valsartan by Solid-State NMR.

    PubMed

    Skotnicki, Marcin; Apperley, David C; Aguilar, Juan A; Milanowski, Bartłomiej; Pyda, Marek; Hodgkinson, Paul

    2016-01-01

    Valsartan (VAL) is an antihypertensive drug marketed in an amorphous form. Amorphous materials can have different physicochemical properties depending on preparation method, thermal history, etc., but the nature of such materials is difficult to study by diffraction techniques. This study characterizes two different amorphous forms of valsartan (AR and AM) using solid-state NMR (SSNMR) as a primary investigation tool, supported by solution-state NMR, FT-IR, TMDSC, and dissolution tests. The two forms are found to be clearly distinct, with a significantly higher level of structural arrangement in the AR form, as observed in (13)C, (15)N, and (1)H SSNMR. (13)C and (15)N NMR indicates that the fully amorphous material (AM) contains an approximately equal ratio of cis-trans conformers about the amide bond, whereas the AR form exists mainly as one conformer, with minor conformational "defects". (1)H ultrafast MAS NMR shows significant differences in the hydrogen bonding involving the tetrazole and acid hydrogens between the two materials, while (15)N NMR shows that both forms exist as a 1,2,3,4-tetrazole tautomer. NMR relaxation times show subtle differences in local and bulk molecular mobility, which can be connected with the glass transition, the stability of the glassy material, and its response to aging. Counterintuitively the fully amorphous material is found to have a significantly lower dissolution rate than the apparently more ordered AR material. PMID:26602457

  18. Motion-adapted pulse sequences for oriented sample (OS) solid-state NMR of biopolymers

    NASA Astrophysics Data System (ADS)

    Lu, George J.; Opella, Stanley J.

    2013-08-01

    One of the main applications of solid-state NMR is to study the structure and dynamics of biopolymers, such as membrane proteins, under physiological conditions where the polypeptides undergo global motions as they do in biological membranes. The effects of NMR radiofrequency irradiations on nuclear spins are strongly influenced by these motions. For example, we previously showed that the MSHOT-Pi4 pulse sequence yields spectra with resonance line widths about half of those observed using the conventional pulse sequence when applied to membrane proteins undergoing rapid uniaxial rotational diffusion in phospholipid bilayers. In contrast, the line widths were not changed in microcrystalline samples where the molecules did not undergo global motions. Here, we demonstrate experimentally and describe analytically how some Hamiltonian terms are susceptible to sample motions, and it is their removal through the critical π/2 Z-rotational symmetry that confers the "motion adapted" property to the MSHOT-Pi4 pulse sequence. This leads to the design of separated local field pulse sequence "Motion-adapted SAMPI4" and is generalized to an approach for the design of decoupling sequences whose performance is superior in the presence of molecular motions. It works by cancelling the spin interaction by explicitly averaging the reduced Wigner matrix to zero, rather than utilizing the 2π nutation to average spin interactions. This approach is applicable to both stationary and magic angle spinning solid-state NMR experiments.

  19. Toxin-induced conformational changes in a potassium channel revealed by solid-state NMR

    NASA Astrophysics Data System (ADS)

    Lange, Adam; Giller, Karin; Hornig, Sönke; Martin-Eauclaire, Marie-France; Pongs, Olaf; Becker, Stefan; Baldus, Marc

    2006-04-01

    The active site of potassium (K+) channels catalyses the transport of K+ ions across the plasma membrane-similar to the catalytic function of the active site of an enzyme-and is inhibited by toxins from scorpion venom. On the basis of the conserved structures of K+ pore regions and scorpion toxins, detailed structures for the K+ channel-scorpion toxin binding interface have been proposed. In these models and in previous solution-state nuclear magnetic resonance (NMR) studies using detergent-solubilized membrane proteins, scorpion toxins were docked to the extracellular entrance of the K+ channel pore assuming rigid, preformed binding sites. Using high-resolution solid-state NMR spectroscopy, here we show that high-affinity binding of the scorpion toxin kaliotoxin to a chimaeric K+ channel (KcsA-Kv1.3) is associated with significant structural rearrangements in both molecules. Our approach involves a combined analysis of chemical shifts and proton-proton distances and demonstrates that solid-state NMR is a sensitive method for analysing the structure of a membrane protein-inhibitor complex. We propose that structural flexibility of the K+ channel and the toxin represents an important determinant for the high specificity of toxin-K+ channel interactions.

  20. Computer-intensive simulation of solid-state NMR experiments using SIMPSON.

    PubMed

    Tošner, Zdeněk; Andersen, Rasmus; Stevensson, Baltzar; Edén, Mattias; Nielsen, Niels Chr; Vosegaard, Thomas

    2014-09-01

    Conducting large-scale solid-state NMR simulations requires fast computer software potentially in combination with efficient computational resources to complete within a reasonable time frame. Such simulations may involve large spin systems, multiple-parameter fitting of experimental spectra, or multiple-pulse experiment design using parameter scan, non-linear optimization, or optimal control procedures. To efficiently accommodate such simulations, we here present an improved version of the widely distributed open-source SIMPSON NMR simulation software package adapted to contemporary high performance hardware setups. The software is optimized for fast performance on standard stand-alone computers, multi-core processors, and large clusters of identical nodes. We describe the novel features for fast computation including internal matrix manipulations, propagator setups and acquisition strategies. For efficient calculation of powder averages, we implemented interpolation method of Alderman, Solum, and Grant, as well as recently introduced fast Wigner transform interpolation technique. The potential of the optimal control toolbox is greatly enhanced by higher precision gradients in combination with the efficient optimization algorithm known as limited memory Broyden-Fletcher-Goldfarb-Shanno. In addition, advanced parallelization can be used in all types of calculations, providing significant time reductions. SIMPSON is thus reflecting current knowledge in the field of numerical simulations of solid-state NMR experiments. The efficiency and novel features are demonstrated on the representative simulations. PMID:25093693

  1. Computer-intensive simulation of solid-state NMR experiments using SIMPSON

    NASA Astrophysics Data System (ADS)

    Tošner, Zdeněk; Andersen, Rasmus; Stevensson, Baltzar; Edén, Mattias; Nielsen, Niels Chr.; Vosegaard, Thomas

    2014-09-01

    Conducting large-scale solid-state NMR simulations requires fast computer software potentially in combination with efficient computational resources to complete within a reasonable time frame. Such simulations may involve large spin systems, multiple-parameter fitting of experimental spectra, or multiple-pulse experiment design using parameter scan, non-linear optimization, or optimal control procedures. To efficiently accommodate such simulations, we here present an improved version of the widely distributed open-source SIMPSON NMR simulation software package adapted to contemporary high performance hardware setups. The software is optimized for fast performance on standard stand-alone computers, multi-core processors, and large clusters of identical nodes. We describe the novel features for fast computation including internal matrix manipulations, propagator setups and acquisition strategies. For efficient calculation of powder averages, we implemented interpolation method of Alderman, Solum, and Grant, as well as recently introduced fast Wigner transform interpolation technique. The potential of the optimal control toolbox is greatly enhanced by higher precision gradients in combination with the efficient optimization algorithm known as limited memory Broyden-Fletcher-Goldfarb-Shanno. In addition, advanced parallelization can be used in all types of calculations, providing significant time reductions. SIMPSON is thus reflecting current knowledge in the field of numerical simulations of solid-state NMR experiments. The efficiency and novel features are demonstrated on the representative simulations.

  2. Solid-State NMR Studies of HIV-1 Capsid Protein Assemblies

    PubMed Central

    Han, Yun; Ahn, Jinwoo; Concel, Jason; Byeon, In-Ja L.; Gronenborn, Angela M.; Yang, Jun; Polenova, Tatyana

    2010-01-01

    In mature HIV-1 virions, a 26.6 kDa CA protein is assembled into a characteristic cone shaped core (capsid) that encloses the RNA viral genome. The assembled capsid structure is best described by a fullerene cone model that is made up from a hexameric lattice containing a variable number of CA pentamers, thus allowing for closure of tubular or conical structures. In this report, we present a solid-state NMR analysis of the wild type HIV-1 CA protein, prepared as conical and spherical assemblies that are stable and are not affected by magic angle spinning of the samples at frequencies between 10 and 25 kHz. Multidimensional homo- and heteronuclear correlation spectra of CA assemblies of uniformly 13C,15N-labelled CA exhibit narrow lines, indicative of conformational homogeneity of the protein in these assemblies. For the conical assemblies, partial residue-specific resonance assignments were obtained. Analysis of the NMR spectra recorded for the conical and spherical assemblies indicates that the CA protein structure is not significantly different in the different morphologies. The present results demonstrate that the assemblies of CA protein are amenable to detailed structural analysis by solid-state NMR spectroscopy. PMID:20092249

  3. Biophysical investigations of membrane perturbations by polypeptides using solid-state NMR spectroscopy (review).

    PubMed

    Bechinger, B

    2000-01-01

    Polypeptides have been prepared by solid-phase peptide synthesis and labelled with 15N at single sites to be used for static or magic angle spinning solid-state NMR spectroscopy. After reconstitution into oriented membranes, the alignment of polypeptide alpha-helices with respect to the bilayer surface is accessible by proton-decoupled 15N solid-state NMR spectroscopy. In addition, limiting values of rotational diffusion coefficients are obtained. The effects of membrane inserted peptides on the bilayer phospholipids have been investigated by 2H and 31P solid-state NMR spectroscopy. Long hydrophobic peptides such as the channel-forming domains of Vpu of HIV-1 or M2 of influenza A adopt stable alignments approximately parallel to the bilayer normal in agreement with models suggesting transmembrane helical bundle formation. The 15N chemical shift data agree with tilt angles of approximately 20 degrees and 33 degrees, respectively. In contrast, multi-charged amphipathic alpha-helices adopt stable orientations parallel to the bilayer surface. In the presence of these peptides, decreased order parameters of the fatty acyl chains, membrane thinning, and the loss of long-range order are observed. Peptides that change topology in a pH dependent manner are more potent in antibiotic assays under experimental conditions where they show in-plane alignments. This result suggests that their detergent-like properties, rather than the formation of transmembrane helical bundles, are responsible for their cell-killing activities. Topological equilibria are also observed within proteins or for polypeptides that do not match the hydrophobic thickness of the bilayer. PMID:11128972

  4. Enlightening the photoactive site of channelrhodopsin-2 by DNP-enhanced solid-state NMR spectroscopy.

    PubMed

    Becker-Baldus, Johanna; Bamann, Christian; Saxena, Krishna; Gustmann, Henrik; Brown, Lynda J; Brown, Richard C D; Reiter, Christian; Bamberg, Ernst; Wachtveitl, Josef; Schwalbe, Harald; Glaubitz, Clemens

    2015-08-11

    Channelrhodopsin-2 from Chlamydomonas reinhardtii is a light-gated ion channel. Over recent years, this ion channel has attracted considerable interest because of its unparalleled role in optogenetic applications. However, despite considerable efforts, an understanding of how molecular events during the photocycle, including the retinal trans-cis isomerization and the deprotonation/reprotonation of the Schiff base, are coupled to the channel-opening mechanism remains elusive. To elucidate this question, changes of conformation and configuration of several photocycle and conducting/nonconducting states need to be determined at atomic resolution. Here, we show that such data can be obtained by solid-state NMR enhanced by dynamic nuclear polarization applied to (15)N-labeled channelrhodopsin-2 carrying 14,15-(13)C2 retinal reconstituted into lipid bilayers. In its dark state, a pure all-trans retinal conformation with a stretched C14-C15 bond and a significant out-of-plane twist of the H-C14-C15-H dihedral angle could be observed. Using a combination of illumination, freezing, and thermal relaxation procedures, a number of intermediate states was generated and analyzed by DNP-enhanced solid-state NMR. Three distinct intermediates could be analyzed with high structural resolution: the early [Formula: see text] K-like state, the slowly decaying late intermediate [Formula: see text], and a third intermediate populated only under continuous illumination conditions. Our data provide novel insight into the photoactive site of channelrhodopsin-2 during the photocycle. They further show that DNP-enhanced solid-state NMR fills the gap for challenging membrane proteins between functional studies and X-ray-based structure analysis, which is required for resolving molecular mechanisms. PMID:26216996

  5. Enlightening the photoactive site of channelrhodopsin-2 by DNP-enhanced solid-state NMR spectroscopy

    PubMed Central

    Becker-Baldus, Johanna; Bamann, Christian; Saxena, Krishna; Gustmann, Henrik; Brown, Lynda J.; Brown, Richard C. D.; Reiter, Christian; Bamberg, Ernst; Wachtveitl, Josef; Schwalbe, Harald; Glaubitz, Clemens

    2015-01-01

    Channelrhodopsin-2 from Chlamydomonas reinhardtii is a light-gated ion channel. Over recent years, this ion channel has attracted considerable interest because of its unparalleled role in optogenetic applications. However, despite considerable efforts, an understanding of how molecular events during the photocycle, including the retinal trans-cis isomerization and the deprotonation/reprotonation of the Schiff base, are coupled to the channel-opening mechanism remains elusive. To elucidate this question, changes of conformation and configuration of several photocycle and conducting/nonconducting states need to be determined at atomic resolution. Here, we show that such data can be obtained by solid-state NMR enhanced by dynamic nuclear polarization applied to 15N-labeled channelrhodopsin-2 carrying 14,15-13C2 retinal reconstituted into lipid bilayers. In its dark state, a pure all-trans retinal conformation with a stretched C14-C15 bond and a significant out-of-plane twist of the H-C14-C15-H dihedral angle could be observed. Using a combination of illumination, freezing, and thermal relaxation procedures, a number of intermediate states was generated and analyzed by DNP-enhanced solid-state NMR. Three distinct intermediates could be analyzed with high structural resolution: the early P1500 K-like state, the slowly decaying late intermediate P4480, and a third intermediate populated only under continuous illumination conditions. Our data provide novel insight into the photoactive site of channelrhodopsin-2 during the photocycle. They further show that DNP-enhanced solid-state NMR fills the gap for challenging membrane proteins between functional studies and X-ray–based structure analysis, which is required for resolving molecular mechanisms. PMID:26216996

  6. Structural modeling of Vpu from HIV-1 based on solid-state NMR observables

    NASA Astrophysics Data System (ADS)

    Ahn, SeonJoo; Lim, GyuTae; Nam, Seungyoon; Lee, Jinhyuk

    2014-04-01

    Vpu is one of the accessory proteins of HIV-1, and is involved in viral particle release. Viral particle release is enhanced by two proposed functions of Vpu: (i) tetherin interaction and (ii) membrane polarization. Two Vpu functions in viral particle release are still controversial. Here, we investigated the proposed functions by extensive structural modeling of Vpu based on solid-state NMR (Nuclear Magnetic Resonance) observables. We found that Vpu can co-exist in two structural forms: left-handed and right-handed conformation. The co-existence of the two conformations provides a clue to reconcile the controversial issue of its two functions in virus particle release.

  7. Solid-state NMR study of geopolymer prepared by sol-gel chemistry

    NASA Astrophysics Data System (ADS)

    Tsai, Yi-Ling; Hanna, John V.; Lee, Yuan-Ling; Smith, Mark E.; Chan, Jerry C. C.

    2010-12-01

    Geopolymers are a new class of materials formed by the condensation of aluminosilicates and silicates obtained from natural minerals or industrial wastes. In this work, the sol-gel method is used to synthesize precursor materials for the preparation of geopolymers. The geopolymer samples prepared by our synthetic route have been characterized by a series of physical techniques, including Fourier-transform infrared, X-ray diffraction, and multinuclear solid-state NMR. The results are very similar to those obtained for the geopolymers prepared from natural kaolinite. We believe that our synthetic approach can offer a good opportunity for the medical applications of geopolymer.

  8. Exploring connections between phase-modulated heteronuclear dipolar decoupling schemes in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Mithu, Venus Singh; Madhu, Perunthiruthy K.

    2013-01-01

    We here show a commonality in the approach to the design of prominent phase-modulated schemes used for heteronuclear dipolar decoupling in solid-state NMR. This concerns mainly two pulse phase modulation (TPPM), small phase incremental alternation (SPINAL), and swept-frequency TPPM (SWf-TPPM) schemes. SPINAL turns out to be a supercycled version of TPPM without any need for phase increments and yielding the same performance. This version of TPPM, called TPPM64, is hence easier to implement than SPINAL. We further compare TPPM64 with SWf-TPPM, another derivative of TPPM which has been shown to perform better than other contemporary heteronuclear dipolar decoupling schemes.

  9. Interference of homonuclear decoupling and exchange in the solid-state NMR of perfluorocyclohexane

    NASA Astrophysics Data System (ADS)

    McMillan, Deborah E.; Hazendonk, Paul; Hodgkinson, Paul

    2003-04-01

    We observe an interference between RF irradiation used for homonuclear decoupling of 19F and conformational exchange in the 13C spectrum of perfluorocyclohexane. We show that these effects can be readily reproduced in simulation, and characterise their dependence on the various NMR and experimental parameters. Their application to observing exchange rates on the kHz timescale is evaluated with respect to T1 ρ measurements and the connections between the two approaches established. The implications for experiments that use homonuclear decoupling of 1H to resolve 1J CH couplings in the solid-state are also evaluated in detail.

  10. Solid-state NMR of the Yersinia pestis outer membrane protein Ail in lipid bilayer nanodiscs sedimented by ultracentrifugation

    PubMed Central

    Ding, Yi; Fujimoto, L. Miya; Yao, Yong; Marassi, Francesca M.

    2015-01-01

    Solid-state NMR studies of sedimented soluble proteins has been recently developed as an attractive approach for overcoming the size limitations of solution NMR spectroscopy and bypassing the need for sample crystallization or precipitation (Bertini et al. 2011). Inspired by the potential benefits of this method, we have investigated the ability to sediment lipid bilayer nanodiscs reconstituted with a membrane protein. In this study, we show that nanodiscs containing the outer membrane protein Ail from Yersinia pestis can be sedimented for solid-state NMR structural studies, without the need for precipitation or lyophilization. Optimized preparations of Ail in phospholipid nanodiscs support both the structure and the fibronectin binding activity of the protein. The same sample can be used for solution NMR, solid-state NMR and activity assays, facilitating structure-activity correlation experiments across a wide range of timescales. PMID:25578899

  11. Proton-Detected Solid-State NMR Spectroscopy of Bone with Ultrafast Magic Angle Spinning

    PubMed Central

    Mroue, Kamal H.; Nishiyama, Yusuke; Kumar Pandey, Manoj; Gong, Bo; McNerny, Erin; Kohn, David H.; Morris, Michael D.; Ramamoorthy, Ayyalusamy

    2015-01-01

    While obtaining high-resolution structural details from bone is highly important to better understand its mechanical strength and the effects of aging and disease on bone ultrastructure, it has been a major challenge to do so with existing biophysical techniques. Though solid-state NMR spectroscopy has the potential to reveal the structural details of bone, it suffers from poor spectral resolution and sensitivity. Nonetheless, recent developments in magic angle spinning (MAS) NMR technology have made it possible to spin solid samples up to 110 kHz frequency. With such remarkable capabilities, 1H-detected NMR experiments that have traditionally been challenging on rigid solids can now be implemented. Here, we report the first application of multidimensional 1H-detected NMR measurements on bone under ultrafast MAS conditions to provide atomistic-level elucidation of the complex heterogeneous structure of bone. Our investigations demonstrate that two-dimensional 1H/1H chemical shift correlation spectra for bone are obtainable using fp-RFDR (finite-pulse radio-frequency-driven dipolar recoupling) pulse sequence under ultrafast MAS. Our results infer that water exhibits distinct 1H−1H dipolar coupling networks with the backbone and side-chain regions in collagen. These results show the promising potential of proton-detected ultrafast MAS NMR for monitoring structural and dynamic changes caused by mechanical loading and disease in bone. PMID:26153138

  12. A Solid-State NMR Study of Selenium Substitution into Nanocrystalline Hydroxyapatite

    PubMed Central

    Kolmas, Joanna; Kuras, Marzena; Oledzka, Ewa; Sobczak, Marcin

    2015-01-01

    The substitution of selenium oxyanions in the hydroxyapatite structure was examined using multinuclear solid-state resonance spectroscopy (ssNMR). The study was supported by powder X-ray diffractometry (PXRD) and wavelength dispersion X-ray fluorescence (WD-XRF). Samples of pure hydroxyapatite (HA300) and selenate (HA300-1.2SeO4) or selenite (HA300-1.2SeO3) substituted hydroxyapatites were synthesized using the standard wet method and heated at 300 °C to remove loosely bonded water. PXRD data showed that all samples are single-phase, nanocrystalline hydroxyapatite. The incorporation of selenite and selenate ions affected the lattice constants. In selenium-containing samples the concentration of Se was very similar and amounted to 9.55% and 9.64%, for HA300-1.2SeO4 and HA300-1.2SeO3, respectively. PXRD and ssNMR data showed that the selenite doping significantly decreases the crystallite size and crystallinity degree. 31P and 1H NMR experiments demonstrated the developed surface hydrated layer in all samples, especially in HA300-1.2SeO3. 1H NMR studies showed the dehydroxylation of HA during the selenium oxyanions substitution and the existence of hydrogen bonding in structural hydroxyl group channels. 1H→77Se cross polarization NMR experiments indicated that selenites and selenates are located in the crystal lattice and on the crystal surface. PMID:25997001

  13. Proton-Detected Solid-State NMR Spectroscopy of Bone with Ultrafast Magic Angle Spinning

    NASA Astrophysics Data System (ADS)

    Mroue, Kamal H.; Nishiyama, Yusuke; Kumar Pandey, Manoj; Gong, Bo; McNerny, Erin; Kohn, David H.; Morris, Michael D.; Ramamoorthy, Ayyalusamy

    2015-07-01

    While obtaining high-resolution structural details from bone is highly important to better understand its mechanical strength and the effects of aging and disease on bone ultrastructure, it has been a major challenge to do so with existing biophysical techniques. Though solid-state NMR spectroscopy has the potential to reveal the structural details of bone, it suffers from poor spectral resolution and sensitivity. Nonetheless, recent developments in magic angle spinning (MAS) NMR technology have made it possible to spin solid samples up to 110 kHz frequency. With such remarkable capabilities, 1H-detected NMR experiments that have traditionally been challenging on rigid solids can now be implemented. Here, we report the first application of multidimensional 1H-detected NMR measurements on bone under ultrafast MAS conditions to provide atomistic-level elucidation of the complex heterogeneous structure of bone. Our investigations demonstrate that two-dimensional 1H/1H chemical shift correlation spectra for bone are obtainable using fp-RFDR (finite-pulse radio-frequency-driven dipolar recoupling) pulse sequence under ultrafast MAS. Our results infer that water exhibits distinct 1H-1H dipolar coupling networks with the backbone and side-chain regions in collagen. These results show the promising potential of proton-detected ultrafast MAS NMR for monitoring structural and dynamic changes caused by mechanical loading and disease in bone.

  14. Homogeneous nanoparticles to enhance the efficiency of a hydrophobic drug, anti-hyperlipidemic Probucol, characterized by solid-state NMR

    PubMed Central

    Io, Takeshi; Fukami, Toshiro; Yamamoto, Kazutoshi; Suzuki, Toyofumi; Xu, Jiadi; Tomono, Kazuo; Ramamoorthy, Ayyalusamy

    2009-01-01

    A low absorption in the gastrointestinal tract of hydrophobic pharmaceutical compounds in use today considerably limits their bioavailability and therefore they are taken in large doses in order to reach the therapeutic plasma concentration, which inevitably results in undesired side effects. In this study, we demonstrate a new nanoparticle approach to overcome this problem and our experimental results show that this approach has a high efficiency of drug loading and is easily adaptable to industrial scale. Characterization of nanoparticles containing a cholesterol-lowering hydrophobic drug, probucol, using a variety of biophysical techniques revealed higher homogeneity of these particles compared to those prepared using other approaches. Intermolecular interactions of these nanoparticles are probed at a high-resolution by magic angle spinning solid-state NMR experiments. PMID:19938876

  15. Restoring Resolution in Biological Solid-State NMR under Conditions of Off-Magic-Angle Spinning.

    PubMed

    Sarkar, Riddhiman; Rodriguez Camargo, Diana C; Pintacuda, Guido; Reif, Bernd

    2015-12-17

    Spin-state-selective excitation (S3E) experiments allow the selection of individual transitions in a coupled two spin system. We show that in the solid state, the dipole-dipole interaction (DD) between (15)N and (1)H in a (1)H-(15)N bond and the chemical shift anisotropy (CSA) of (15)N in an amide moiety mutually cancel each other for a particular multiplet component at high field, when the sample is spun off the magic angle (Arctan [√2] = 54.74°). The accuracy of the adjustment of the spinning angle is crucial in conventional experiments. We demonstrate that for S3E experiments, the requirement to spin the sample exactly at the magic angle is not mandatory. Applications of solid state NMR in narrow bore magnets will be facilitated where the adjustment of the magic angle is often difficult. The method opens new perspectives for the development of schemes to determine distances and to quantify dynamics in the solid state. PMID:26641130

  16. Advances in solid-state relaxation methodology for probing site-specific protein dynamics.

    PubMed

    Lewandowski, Józef Romuald

    2013-09-17

    Dynamics are intimately linked to protein stability and play a crucial role in important biological processes, such as ligand binding, allosteric regulation, protein folding, signaling, and enzymatic catalysis. Solid-state NMR relaxation measurements allow researchers to determine the amplitudes, time scales, and under favorable conditions, directionality of motions at atomic resolution over the entire range of dynamic processes from picoseconds to milliseconds. Because this method allows researchers to examine both the amplitudes and time scales of motions in this range, they can link different tiers of protein motions in protein energy landscapes. As a result, scientists can better understand the relationships between protein motions and functions. Such studies are possible both with the primary targets of solid-state NMR studies, such as amyloid fibrils, membrane proteins, or other heterogeneous systems, and others that researchers typically study by solution NMR and X-ray crystallography. In addition, solid-state NMR, with the absence of tumbling in solution, eliminates the intrinsic size limitation imposed by slow tumbling of large proteins. Thus, this technique allows researchers to characterize interdomain and intermolecular interactions in large complexes at the atomic scale. In this Account, we discuss recent advances in solid-state relaxation methodology for studying widespread site-specific protein dynamics. We focus on applications involving magic angle spinning, one of the primary methods used in high-resolution solid-state NMR. We give an overview of challenges and solutions for measuring (15)N and (13)C spin-lattice relaxation (R1) to characterize fast picosecond-nanosecond motions, spin-lattice in the rotating frame (R1ρ), and other related relaxation rates for characterization of picosecond-millisecond protein motions. In particular, we discuss the problem of separating incoherent effects caused by random motions from coherent effects arising from incomplete averaging of orientation-dependent NMR interactions. We mention a number of quantitative studies of protein dynamics based on solid-state relaxation measurements. Finally, we discuss the potential use of relaxation measurements for extracting the directionality of motions. Using the (15)N and (13)C R1 and R1ρ measurements, we illustrate the backbone and side-chain dynamics in the protein GB1 and comment on this emerging dynamic picture within the context of data from solution NMR measurements and simulations. PMID:23621579

  17. Quantitative structural characterization of POSS and octavinyl-POSS nanocomposites by solid state NMR.

    PubMed

    Zhao, Huipeng; Shu, Jie; Chen, Qun; Zhang, Shanmin

    2012-01-01

    The ratio between two different (29)Si atoms in chloromethylphenyl isobutyl Polyhedral Oligomeric Silsesquioxane (POSS) was determined based on the quantitative cross polarization (QCP) (Shu et al., Chem. Phys. Lett. 462 (2008) 125) in solid-state NMR. For a (29)Si/(1)H spin system, cross polarization and depolarization together with the reciprocity relation were performed with optimized experimental conditions. It saves considerable experimental time compared to the (29)Si direct polarization experiment. The same method was further applied to octavinyl-POSS nanocomposites containing perfluoropolyether (PFPE) for deriving directly and accurately the average numbers of reacted vinyl groups, which may not be obtained by combining FTIR and solution (1)H NMR. In principle, the aforementioned method proves to be valuable in quantitative characterization of silicon related structures in bulk materials. PMID:22503875

  18. The rehydration of metakaolinite to kaolinite: Evidence from solid-state NMR and cognate techniques

    SciTech Connect

    Rocha, J.; Klinowski, J. ); Adams, J.M. )

    1990-12-01

    The authors have studied the process of rehydroxylation of metakaolinite to kaolinite by {sup 27}Al and {sup 29}Si MAS NMR in tandem with Fourier transform infrared spectroscopy, powder X-ray diffraction, thermogravimetric analysis, specific surface area measurement, atomic absorption spectrophotometry, and electron microscopy. The efficiency of the process strongly depends on temperature. The reaction is initiated at the edges of metakaolinite particles and is followed by diffusion of the water into their bulk. Regions of the metakaolinite structure, which locally retain the symmetry of kaolinite, act as nuclei for the reconstitution of kaolinite particles. The latter grown perpendicular to the c{sup *} axis and their edges are parallel to those of metakaolinite, showing that rehydroxylation is a topotactic solid-state process. {sup 27}Al and {sup 29}Si MAS NMR reveal that metakaolinite is an ill-defined mixture of amorphous silica nd alumina, and can be transformed into kaolinite by a correct choice of experimental conditions.

  19. Solid state 13C NMR characterisation study on fourth generation Ziegler-Natta catalysts.

    PubMed

    Heikkinen, Harri; Liitiä, Tiina; Virkkunen, Ville; Leinonen, Timo; Helaja, Tuulamari; Denifl, Peter

    2012-01-01

    In this study, solid state (13)C NMR spectroscopy was utilised to characterize and identify the metal-ester coordination in active fourth generation (phthalate) Ziegler-Natta catalysts. It is known that different donors affect the active species in ZN catalysts. However, there is still limited data available of detailed molecular information how the donors and the active species are interplaying. One of the main goals of this work was to get better insight into the interactions of donor and active species. Based on the anisotropy tensor values (δ(11), δ(22), δ(33)) from low magic-angle spinning (MAS) (13)C NMR spectra in combination with chemical shift anisotropy (CSA) calculations (δ(aniso) and η), both the coordinative metal (Mg/Ti) and the symmetry of this interaction between metal and the internal donor in the active catalyst (MgCl(2)/TiCl(4)/electron donor) system could be identified. PMID:22425229

  20. β-Helical architecture of cytoskeletal bactofilin filaments revealed by solid-state NMR

    PubMed Central

    Vasa, Suresh; Lin, Lin; Shi, Chaowei; Habenstein, Birgit; Riedel, Dietmar; Kühn, Juliane; Thanbichler, Martin; Lange, Adam

    2015-01-01

    Bactofilins are a widespread class of bacterial filament-forming proteins, which serve as cytoskeletal scaffolds in various cellular pathways. They are characterized by a conserved architecture, featuring a central conserved domain (DUF583) that is flanked by variable terminal regions. Here, we present a detailed investigation of bactofilin filaments from Caulobacter crescentus by high-resolution solid-state NMR spectroscopy. De novo sequential resonance assignments were obtained for residues Ala39 to Phe137, spanning the conserved DUF583 domain. Analysis of the secondary chemical shifts shows that this core region adopts predominantly β-sheet secondary structure. Mutational studies of conserved hydrophobic residues located in the identified β-strand segments suggest that bactofilin folding and polymerization is mediated by an extensive and redundant network of hydrophobic interactions, consistent with the high intrinsic stability of bactofilin polymers. Transmission electron microscopy revealed a propensity of bactofilin to form filament bundles as well as sheet-like, 2D crystalline assemblies, which may represent the supramolecular arrangement of bactofilin in the native context. Based on the diffraction pattern of these 2D crystalline assemblies, scanning transmission electron microscopy measurements of the mass per length of BacA filaments, and the distribution of β-strand segments identified by solid-state NMR, we propose that the DUF583 domain adopts a β-helical architecture, in which 18 β-strand segments are arranged in six consecutive windings of a β-helix. PMID:25550503

  1. Solid-state NMR analyses of the structure and dynamics of polymers in the different states

    NASA Astrophysics Data System (ADS)

    Horii, Fumitaka; Kaji, Hironori; Ishida, Hiroyuki; Kuwabara, Kazuhiro; Masuda, Kenji; Tai, Toshihiro

    1998-01-01

    This paper reviews our recent investigations of the structure and molecular motions of synthetic polymers in the different states, which were mainly carried out by solid-state 13C or 2H NMR spectroscopy. First, the crystalline-noncrystalline structure is characterized in detail for metallocene-catalyzed linear low density polyethylene, which was isothermally crystallized from the melt. Secondly, the characteristic chain conformation, the alternate trans and rapid trans-gauche exchange conformation, is revealed for the spacer methylene sequence in the frozen liquid crystalline region for thermotropic liquid crystalline polyurethane, which was crystallized from the isotropic phase through the nematic liquid crystalline phase. Thirdly, different types of splittings of CH resonance lines, which will be induced mainly by the formation of intramolecular hydrogen bonds in the triad sequence, are shown in the frozen solution states for poly(vinyl alcohol) samples with different tacticities. It is finally pointed out by solid-state 2H NMR analyses that both distributions in flip frequency and in flip angle should be taken into account for the precise characterization of the phenylene flip motion in glassy poly(ethylene terephthalate).

  2. β-Helical architecture of cytoskeletal bactofilin filaments revealed by solid-state NMR.

    PubMed

    Vasa, Suresh; Lin, Lin; Shi, Chaowei; Habenstein, Birgit; Riedel, Dietmar; Kühn, Juliane; Thanbichler, Martin; Lange, Adam

    2015-01-13

    Bactofilins are a widespread class of bacterial filament-forming proteins, which serve as cytoskeletal scaffolds in various cellular pathways. They are characterized by a conserved architecture, featuring a central conserved domain (DUF583) that is flanked by variable terminal regions. Here, we present a detailed investigation of bactofilin filaments from Caulobacter crescentus by high-resolution solid-state NMR spectroscopy. De novo sequential resonance assignments were obtained for residues Ala39 to Phe137, spanning the conserved DUF583 domain. Analysis of the secondary chemical shifts shows that this core region adopts predominantly β-sheet secondary structure. Mutational studies of conserved hydrophobic residues located in the identified β-strand segments suggest that bactofilin folding and polymerization is mediated by an extensive and redundant network of hydrophobic interactions, consistent with the high intrinsic stability of bactofilin polymers. Transmission electron microscopy revealed a propensity of bactofilin to form filament bundles as well as sheet-like, 2D crystalline assemblies, which may represent the supramolecular arrangement of bactofilin in the native context. Based on the diffraction pattern of these 2D crystalline assemblies, scanning transmission electron microscopy measurements of the mass per length of BacA filaments, and the distribution of β-strand segments identified by solid-state NMR, we propose that the DUF583 domain adopts a β-helical architecture, in which 18 β-strand segments are arranged in six consecutive windings of a β-helix. PMID:25550503

  3. Broadband Heteronuclear Solid-State NMR Experiments by Exponentially Modulated Dipolar Recoupling without Decoupling

    PubMed Central

    Nielsen, Anders B.; Straasø, Lasse A.; Nieuwkoop, Andrew J.; Rienstra, Chad M.; Bjerring, Morten; Nielsen, Niels Chr.

    2010-01-01

    We present a novel solid-state NMR method for heteronuclear dipolar recoupling without decoupling. The method, which introduces the concept of exponentially modulated rf fields, provides efficient broadband recoupling with large flexibility with respect to hetero- or homonuclear applications, sample spinning frequency, and operation without the need for high-power 1H decoupling. For previous methods, the latter has been a severe source of sample heating which may cause detoriation of costly samples. The so-called EXPonentially mOdulated Recoupling Technique (EXPORT) is described analytically and numerically, and demonstrated experimentally by 1D 13C spectra and 2D 13C-15N correlation spectra of 13C,15N-labeled samples of GB1, ubiquitin, and fibrils of the SNNFGAILSS fragment of amylin. Through its flexible operation, robustness, and strong performance, it is anticipated that EXPORT will find immediate application for both hetero- and homonuclear dipolar recoupling in solid-state NMR of 13C,15N-labeled proteins and compounds of relevance in chemistry. PMID:20689682

  4. Chemical structure and heterogeneity differences of two lignins from loblolly pine as investigated by advanced solid-state NMR spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Advanced solid-state NMR was employed to investigate differences in chemical structure and heterogeneity between milled wood lignin (MWL) and residual enzyme lignin (REL). Wiley and conventional milled woods were also studied. The advanced NMR techniques included 13C quantitative direct polarization...

  5. Experimental aspects in acquisition of wide bandwidth solid-state MAS NMR spectra of low-? nuclei with different opportunities on two commercial NMR spectrometers.

    PubMed

    Jakobsen, Hans J; Bildse, Henrik; Gan, Zhehong; Brey, William W

    2011-08-01

    The acquisition and different appearances observed for wide bandwidth solid-state MAS NMR spectra of low-? nuclei, using (14)N as an illustrative nucleus and employing two different commercial spectrometers (Varian, 14.1T and Bruker, 19.6T), have been compared/evaluated and optimized from an experimental NMR and an electronic engineering point of view, to account for the huge differences in these spectra. The large differences in their spectral appearances, employing the recommended/standard experimental set-up for the two different spectrometers, are shown to be associated with quite large differences in the electronic design of the two types of preamplifiers, which are connected to their respective probes through a 50? cable, and are here completely accounted for. This has led to different opportunities for optimum performances in the acquisition of nearly ideal wide bandwidth spectra for low-? nuclei on the two spectrometers by careful evaluation of the length for the 50? probe-to-preamp cable for the Varian system and appropriate changes to the bandwidth (Q) of the NMR probe used on the Bruker spectrometer. Earlier, we reported quite distorted spectra obtained with Varian Unity INOVA spectrometers (at 11.4 and 14.1T) in several exploratory wide bandwidth (14)N MAS NMR studies of inorganic nitrates and amino acids. These spectra have now been compared/evaluated with fully analyzed (14)N MAS spectra correspondingly acquired at 19.6T on a Bruker spectrometer. It is shown that our upgraded version of the STARS simulation/iterative-fitting software is capable of providing identical sets for the molecular spectral parameters and corresponding fits to the experimental spectra, which fully agree with the electronic measurements, despite the highly different appearances for the MAS NMR spectra acquired on the Varian and Bruker spectrometers. PMID:21704544

  6. Combining solid-state NMR spectroscopy with first-principles calculations - a guide to NMR crystallography.

    PubMed

    Ashbrook, Sharon E; McKay, David

    2016-06-01

    Recent advances in the application of first-principles calculations of NMR parameters to periodic systems have resulted in widespread interest in their use to support experimental measurement. Such calculations often play an important role in the emerging field of "NMR crystallography", where NMR spectroscopy is combined with techniques such as diffraction, to aid structure determination. Here, we discuss the current state-of-the-art for combining experiment and calculation in NMR spectroscopy, considering the basic theory behind the computational approaches and their practical application. We consider the issues associated with geometry optimisation and how the effects of temperature may be included in the calculation. The automated prediction of structural candidates and the treatment of disordered and dynamic solids are discussed. Finally, we consider the areas where further development is needed in this field and its potential future impact. PMID:27117884

  7. Solid-State Ru-99 NMR Spectroscopy: A Useful Tool for Characterizing Prototypal Diamagnetic Ruthenium Compounds Compounds

    SciTech Connect

    Ooms, Kristopher J.; Wasylishen, Roderick E.

    2004-09-08

    The feasibility of 99Ru NMR spectroscopy as a tool to characterize solid compounds is demonstrated. Results of the first solid-state 99Ru NMR investigation of diamagnetic compounds are presented for Ru(NH3)6Cl2, K4Ru(CN)6 xH2O (x) 0, 3, LaKRu(CN)6, and Ru3(CO)12. The sensitivity of the ruthenium magnetic shielding tensor to subtle changes in the local structure about the ruthenium nucleus is highlighted by comparing the 99Ru isotropic chemical shift of Ru(NH3)6Cl2 in aqueous solutions and in the solid state. The narrow isotropic 99Ru NMR peak observed for solid Ru(NH3)6Cl2 indicates that this compound is an ideal secondary reference sample for solid-state 99Ru NMR studies.

  8. Molecular structure of crude beeswax studied by solid-state 13C NMR

    PubMed Central

    Kameda, Tsunenori

    2004-01-01

    13C Solid-state NMR experiments were performed to investigate the structure of beeswax in the native state (crude beeswax) for the first time. From quantitative direct polarization 13C MAS NMR spectrum, it was found that the fraction of internal-chain methylene (int-(CH2)) component compared to other components of crude beeswax was over 95%. The line shape of the int-(CH2) carbon resonance region was comprehensively analyzed in terms of NMR chemical shift. The 13C broad peak component covering from 31 to 35ppm corresponds to int-(CH2) carbons with trans conformation in crystalline domains, whereas the sharp signal at 30.3 ppm corresponds to gauche conformation in the non-crystalline domain. From peak deconvolution of the aliphatic region, it was found that over 85% of the int-(CH2) has a crystal structure and several kinds of molecular packing for int-(CH2), at least three, exist in the crystalline domain. Abbreviation: NMR nuclear magnetic resonance int-(CH2) internal-chain methylene CP cross-polarization MAS magic angle spinning PMID:15861244

  9. Solid-state and unilateral NMR study of deterioration of a Dead Sea Scroll fragment.

    PubMed

    Masic, A; Chierotti, M R; Gobetto, R; Martra, G; Rabin, I; Coluccia, S

    2012-02-01

    Unilateral and solid-state nuclear magnetic resonance (NMR) analyses were performed on a parchment fragment of the Dead Sea Scroll (DSS). The analyzed sample belongs to the collection of non-inscribed and nontreated fragments of known archaeological provenance from the John Rylands University Library in Manchester. Therefore, it can be considered as original DSS material free from any contamination related to the post-discovery period. Considering the paramount significance of the DSS, noninvasive approaches and portable in situ nondestructive methods are of fundamental importance for the determination of composition, structure, and chemical-physical properties of the materials under study. NMR studies reveal low amounts of water content associated with very short proton relaxation times, T(1), indicating a high level of deterioration of collagen molecules within scroll fragments. In addition, (13)C cross-polarization magic-angle-spinning (CPMAS) NMR spectroscopy shows characteristic peaks of lipids whose presence we attribute to the production technology that did not involve liming. Extraction with chloroform led to the reduction of both lipid and protein signals in the (13)C CPMAS spectrum indicating probable involvement of lipids in parchment degradation processes. NMR absorption and relaxation measurements provide nondestructive, discriminative, and sensitive tools for studying the deterioration effects on the organization and properties of water and collagen within ancient manuscripts. PMID:21805064

  10. Quantitative solid-state 13C NMR with signal enhancement by multiple cross polarization.

    PubMed

    Johnson, Robert L; Schmidt-Rohr, Klaus

    2014-02-01

    A simple new method is presented that yields quantitative solid-state magic-angle spinning (MAS) (13)C NMR spectra of organic materials with good signal-to-noise ratios. It achieves long (>10ms) cross polarization (CP) from (1)H without significant magnetization losses due to relaxation and with a moderate duty cycle of the radio-frequency irradiation, by multiple 1-ms CP periods alternating with (1)H spin-lattice relaxation periods that repolarize the protons. The new method incorporates previous techniques that yield less distorted CP/MAS spectra, such as a linear variation ("ramp") of the radio-frequency field strength, and it overcomes their main limitation, which is T1ρ relaxation of the spin-locked (1)H magnetization. The ramp of the radio-frequency field strength and the asymptotic limit of cross polarization makes the spectral intensity quite insensitive to the exact field strengths used. The new multiCP pulse sequence is a "drop-in" replacement for previous CP methods and produces no additional data-processing burden. Compared to the only reliable quantitative (13)C NMR method for unlabeled solids previously available, namely direct-polarization NMR, the measuring time is reduced by more than a factor of 50, enabling higher-throughput quantitative NMR studies. The new multiCP technique is validated with 14-kHz MAS on amino-acid derivatives, plant matter, a highly aromatic humic acid, and carbon materials made by low-temperature pyrolysis. PMID:24374751

  11. Probing access resistance of solid-state nanopores with a scanning-probe microscope tip.

    PubMed

    Hyun, Changbae; Rollings, Ryan; Li, Jiali

    2012-02-01

    An apparatus that integrates solid-state nanopore ionic current measurement with a scanning-probe microscope is developed. When a micrometer-scale scanning-probe tip is near a voltage-biased nanometer-scale pore (10–100 nm), the tip partially blocks the flow of ions to the pore and increases the pore access resistance. The apparatus records the current blockage caused by the probe tip and the location of the tip simultaneously. By measuring the current blockage map near a nanopore as a function of the tip position in 3D space in salt solution, the relative pore resistance increases due to the tip and ΔR/R0 is estimated as a function of the tip location, nanopore geometry, and salt concentration. The amplitude of ΔR/R0 also depends on the ratio of the pore length to its radius as Ohm's law predicts. When the tip is very close to the pore surface, ≈10 nm, experiments show that ΔR/R0 depends on salt concentration as predicted by the Poisson and Nernst–Planck equations. Furthermore, the measurements show that ΔR/R0 goes to zero when the tip is about five times the pore diameter away from the center of the pore entrance. The results in this work not only demonstrate a way to probe the access resistance of nanopores experimentally; they also provide a way to locate the nanopore in salt solution, and open the door to future nanopore experiments for detecting single biomolecules attached to a probe tip. PMID:22287084

  12. Probing Access Resistance of Solid-state Nanopores with a Scanning Probe Microscope Tip.

    PubMed

    Hyun, Changbae; Rollings, Ryan; Li, Jiali

    2012-02-01

    An apparatus that integrates solid-state nanopore ionic current measurement with a Scanning Probe Microscope has been developed. When a micrometer-scale scanning probe tip is near a voltage biased nanometer-scale pore (10-100 nm), the tip partially blocks the flow of ions to the pore and increases the pore access resistance. The apparatus records the current blockage caused by the probe tip and the location of the tip simultaneously. By measuring the current blockage map near a nanopore as a function of the tip position in 3D space in salt solution, we estimate the relative pore resistance increase due to the tip, ΔR/R(0), as a function of the tip location, nanopore geometry, and salt concentration. The amplitude of ΔR/R(0) also depends on the ratio of the pore length to its radius as Ohm's law predicts. When the tip is very close to the pore surface, ~10 nm, our experiments show that ΔR/R(0) depends on salt concentration as predicted by the Poisson and Nernst-Planck equations. Furthermore, our measurements show that ΔR/R(0) goes to zero when the tip is about five times the pore diameter away from the center of the pore entrance. The results in this work not only demonstrate a way to probe the access resistance of nanopores experimentally, they also provide a way to locate the nanopore in salt solution, and open the door to future nanopore experiments for detecting single biomolecules attached to a probe tip. PMID:22393313

  13. Applications of Solid State NMR to the Study of Molecular Structure

    NASA Astrophysics Data System (ADS)

    Curtis, Ronald Dean

    This thesis illustrates several applications of dilute spin I = 1over2 solid state nmr spectroscopy to the study of molecular structure in systems of chemical interest. Specifically, the compounds studied include benzylideneaniline and several related imines, the first stable iminophosphenium cation containing a N,P triple bond and several tetracyclines. The first two applications describe the use of dipolar-chemical shift nmr of "isolated" spin-pairs to fully characterize chemical shift tensors. For example, the carbon and nitrogen shift tensors of the C=N linkage of the Schiff base benzylideneaniline have been completely specified. The most shielded principal component of both carbon and nitrogen shift tensors is approximately perpendicular to the imine fragment. For the imine carbon, the intermediate component of the shift tensor is directed approximately along the C=N bond whereas the corresponding component of the nitrogen shift tensor is oriented along the direction of the nitrogen lone pair. Examination of the nitrogen chemical shift parameters for several related imines suggests that variations in the least shielded principal component are mainly responsible for changes in the nitrogen shieldings in the imine system. For the N,P moiety of the iminophosphenium cation, the most shielded principal component of both nitrogen and phosphorus tensors is oriented along the N,P bond axis. Comparison of both shift tensors with those of related compounds suggests that the electronic environment surrounding the N,P moiety is similar to other systems containing a formal triple bond. The final application section demonstrates the utility of high-resolution ^{13} C and ^{15}N cp/mas nmr for studying the molecular structure of solid tetracycline antibiotics. Comparison of ^{15} C chemical shifts in the solid state to those determined in (CD_3)_2SO solutions indicates for the first time that the structural integrity of the A ring of the tetracyclines is maintained in solution.

  14. Solid-state NMR analysis of membrane proteins and protein aggregates by proton detected spectroscopy

    PubMed Central

    Nieuwkoop, Andrew J.; Berthold, Deborah A.; Comellas, Gemma; Sperling, Lindsay J.; Tang, Ming; Shah, Gautam J.; Brea, Elliott J.; Lemkau, Luisel R.

    2012-01-01

    Solid-state NMR has emerged as an important tool for structural biology and chemistry, capable of solving atomic-resolution structures for proteins in membrane-bound and aggregated states. Proton detection methods have been recently realized under fast magic-angle spinning conditions, providing large sensitivity enhancements for efficient examination of uniformly labeled proteins. The first and often most challenging step of protein structure determination by NMR is the site-specific resonance assignment. Here we demonstrate resonance assignments based on high-sensitivity proton-detected three-dimensional experiments for samples of different physical states, including a fully-protonated small protein (GB1, 6 kDa), a deuterated microcrystalline protein (DsbA, 21 kDa), a membrane protein (DsbB, 20 kDa) prepared in a lipid environment, and the extended core of a fibrillar protein (α-synuclein, 14 kDa). In our implementation of these experiments, including CONH, CO(CA)NH, CANH, CA(CO)NH, CBCANH, and CBCA(CO)NH, dipolar-based polarization transfer methods have been chosen for optimal efficiency for relatively high protonation levels (full protonation or 100 % amide proton), fast magic-angle spinning conditions (40 kHz) and moderate proton decoupling power levels. Each H–N pair correlates exclusively to either intra- or inter-residue carbons, but not both, to maximize spectral resolution. Experiment time can be reduced by at least a factor of 10 by using proton detection in comparison to carbon detection. These high-sensitivity experiments are especially important for membrane proteins, which often have rather low expression yield. Proton-detection based experiments are expected to play an important role in accelerating protein structure elucidation by solid-state NMR with the improved sensitivity and resolution. PMID:22986689

  15. Molecular-level characterization of probucol nanocrystal in water by in situ solid-state NMR spectroscopy.

    PubMed

    Zhang, Junying; Higashi, Kenjirou; Limwikrant, Waree; Moribe, Kunikazu; Yamamoto, Keiji

    2012-02-28

    The molecular state of colloidal probucol nanoparticles with additives was evaluated by (13)C in situ solid-state NMR spectroscopy. The nanoparticles were obtained by dispersing a ternary co-ground mixture of probucol/polyvinylpyrrolidon (PVP)/sodium dodecyl sulfate (SDS) in water. Their mean particle size was found to be approximately 150 nm by dynamic light scattering and cryogenic-scanning electron microscopy measurements. The results of the (13)C in situ solid-state NMR spectroscopy showed that probucol existed in the crystalline state (form I) in water. (13)C liquid-state NMR results indicated that PVP and SDS interacted with probucol in water. Their broad signals suggested that the surface interaction of the probucol nanocrystal with PVP and SDS stabilized the suspension. In addition, a freeze-dried sample of the suspension was studied by (13)C solid-state NMR and powder X-ray diffraction experiments, which confirmed the presence of the probucol nanocrystals. The combination of the in situ solid-state, solid-state, and liquid-state NMR measurement results provided molecular-level insights about the role of intermolecular interactions in the design of nanoformulations. PMID:22138607

  16. Solid state NMR study of SEI formation in lithium ion batteries

    NASA Astrophysics Data System (ADS)

    Zhao, Dachun

    Recently, rechargeable lithium ion batteries, which offer high energy density and long cycle life, are in great demand as power sources for our mobile electronic society. The formation of a solid electrolyte interphase (SEI) on the surface of electrodes in lithium ion batteries plays an essential role in their performance. This thesis presents solid state NMR and MAS NMR results on the SEI, which contribute to our understanding of SEI formation on both cathodes and anodes. This thesis is organized as following: Chapter 1 surveys the history of batteries and the challenges to further development of the lithium ion battery. Fundamental aspects and SEI formation mechanisms are also included in Chapter l. Chapter 2 deals with the principles and experimental techniques of solid state NMR. Chapter 3 presents studies of SEI formation on anode and cathode in lithium ion batteries using electrochemical impedance spectroscopy (EIS) and NMR. The results provide EIS and NMR evidence that cells containing electrolytes with high EC content display less irreversible capacity after high temperature storage. The irreversible capacity is attributed to SEI growth on electrode surfaces. NMR results on cathodes, on the other hand, imply that the presence of Ni in the cathode may reduce cell performance due to the oxidation of Ni 3+ to Ni4+. Our simulations show that a lower EC/DMC ratio is associated with a smaller SEI intensity for the cathode and higher intensity for the anode. Chapter 4 discusses the effect of temperature on SEI formation on anodes and cathodes. NMR measurements show that MCMB graphite based anodes exhibit high stability no chemical shift is evident over a wide temperature range. On cathodes, however, NMR does reveal changes in SEI intensity as a function of temperature. These changes are believed to be the result of decomposition of the SEI. Evidently, then, changes in the performance of the cell as a factor of temperature are, at least in part, due to changes in the SEI with temperature. In Chapter 5 we report on the use of NMR to study the effect of electrolyte/solvent and electrode structure on SEI formation. The intensity of the SEI in a cell containing LiBOB electrolyte with EC and DEC is greater than that in a cell with LiPF6 electrolyte with EC and DMC. This implies that the cell containing LiBOB electrolyte with EC and DEC is more stable. From the analysis of NMR of the cathode, it was found that it is easier to form SEI in layered structure cathode than in a spinel structure, suggesting that spinel structure materials may be better choices as cathode in lithium ion batteries. Evidently, then, the choice of electrolyte/solvent materials and electrode materials are key factors in determining the performance of lithium ion batteries.

  17. Action of the multifunctional peptide BP100 on native biomembranes examined by solid-state NMR.

    PubMed

    Misiewicz, Julia; Afonin, Sergii; Grage, Stephan L; van den Berg, Jonas; Strandberg, Erik; Wadhwani, Parvesh; Ulrich, Anne S

    2015-04-01

    Membrane composition is a key factor that regulates the destructive activity of antimicrobial peptides and the non-leaky permeation of cell penetrating peptides in vivo. Hence, the choice of model membrane is a crucial aspect in NMR studies and should reflect the biological situation as closely as possible. Here, we explore the structure and dynamics of the short multifunctional peptide BP100 using a multinuclear solid-state NMR approach. The membrane alignment and mobility of this 11 amino acid peptide was studied in various synthetic lipid bilayers with different net charge, fluidity, and thickness, as well as in native biomembranes harvested from prokaryotic and eukaryotic cells. (19)F-NMR provided the high sensitivity and lack of natural abundance background that are necessary to observe a labelled peptide even in protoplast membranes from Micrococcus luteus and in erythrocyte ghosts. Six selectively (19)F-labeled BP100 analogues gave remarkably similar spectra in all of the macroscopically oriented membrane systems, which were studied under quasi-native conditions of ambient temperature and full hydration. This similarity suggests that BP100 has the same surface-bound helical structure and high mobility in the different biomembranes and model membranes alike, independent of charge, thickness or cholesterol content of the system. (31)P-NMR spectra of the phospholipid components did not indicate any bilayer perturbation, so the formation of toroidal wormholes or micellarization can be excluded as a mechanism of its antimicrobial or cell penetrating action. However, (2)H-NMR analysis of the acyl chain order parameter profiles showed that BP100 leads to considerable membrane thinning and thereby local destabilization. PMID:25616492

  18. Application of Solid-State NMR Relaxometry for Characterization and Formulation Optimization of Grinding-Induced Drug Nanoparticle.

    PubMed

    Ueda, Keisuke; Higashi, Kenjirou; Moribe, Kunikazu

    2016-03-01

    The formation mechanism of drug nanoparticles was investigated using solid-state nuclear magnetic resonance (NMR) techniques for the efficient discovery of an optimized nanoparticle formulation. The cogrinding of nifedipine (NIF) with polymers, including hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP), and sodium dodecyl sulfate (SDS) was performed to prepare the NIF nanoparticle formulations. Then, solid-state NMR relaxometry was used for the nanometer-order characterization of NIF in the polymer matrix. Solid-state NMR measurements revealed that the crystal size of NIF was reduced to several tens of nanometers with amorphization of NIF by cogrinding with HPMC and SDS for 100 min. Similarly, the size of the NIF crystal was reduced to less than 90 nm in the 40 min ground mixture of NIF/PVP/SDS. Furthermore, 100 min grinding of NIF/PVP/SDS induced amorphization of almost all the NIF crystals followed by nanosizing. The hydrogen bond between NIF and PVP led to the efficient amorphization of NIF in the NIF/PVP/SDS system compared with NIF/HPMC/SDS system. The efficient nanosizing of the NIF crystal in the solid state, revealed by the solid-state NMR relaxation time measurements, enabled the formation of large amounts of NIF nanoparticles in water followed by the polymer dissolution. In contrast, excess amorphization of the NIF crystals failed to efficiently prepare the NIF nanoparticles. The solid-state characterization of the crystalline NIF revealed good correlation with the NIF nanoparticles formation during aqueous dispersion. Furthermore, the solid-state NMR measurements including relaxometry successfully elucidated the nanometer-order dispersion state of NIF in polymer matrix, leading to the discovery of optimized conditions for the preparation of suitable drug nanoparticles. PMID:26855230

  19. Molecular ordering of mixed surfactants in mesoporous silicas: A solid-state NMR study

    SciTech Connect

    Kobayashi, Takeshi; Mao, Kanmi; Wang, Shy-Guey; Lin, Victor S.-Y.; Pruski, Marek

    2011-02-17

    The use of mixed surfactants in the synthesis of mesoporous silica nanoparticles (MSNs) is of importance in the context of adjusting pore structures, sizes and morphologies. In the present study, the arrangement of molecules in micelles produced from a mixture of two surfactants, cetyltrimethylammonium bromide (CTAB) and cetylpyridinium bromide (CPB) was detailed by solid-state NMR spectroscopy. Proximities of methyl protons in the trimethylammonium headgroup of CTAB and protons in the pyridinium headgroup of CPB were observed under fast magic angle spinning (MAS) by {sup 1}H-{sup 1}H double quantum (DQ) MAS NMR and NOESY. This result suggested that CTAB and CPB co-exist in the pores without forming significant monocomponent domain structures. {sup 1}H-{sup 29}Si heteronuclear correlation (HETCOR) NMR showed that protons in the headgroups of CTAB are in closer proximity to the silica surface than those in the CPB headgroups. The structural information obtained in this investigation leads to better understanding of the mechanisms of self-assembly and their role in determining the structure and morphology of mesoporous materials.

  20. Solid-state NMR studies of the reactions of silica surfaces with polyfunctional chloromethylsilanes and ethoxymethylsilanes

    SciTech Connect

    Sindorf, D.W.; Maciel, G.E.

    1983-06-15

    The silylation of silica (Si) surfaces by dimethyldichlorosilane, methyltrichlorosilane, dimethyldiethoxysilane, and methyltriethoxysilane has been studied by solid-state /sup 29/Si and /sup 13/C NMR, by use of cross polarization (CP) and magic-angle spinning (MAS). An earlier formalism for the quantitative analysis of the NRM data has been extended for use with polyfunctional silyating agents and applied in detail to dimethyldichlorosilane reactions. Silylation with ethoxysilane reagents is less amenable to quantitative interpretation but appears to parallel closely the behavior of analogous chlorosilane reagents. With ethoxysilane reagents /sup 13/C NMR is found to be quite useful, especially for determining the fate of ethoxy groups. In all of the cases studied, the products are characterized primarily by single silane-to-surface attachments of each silane silicon atom. The presence of absorbed water is found to play an important role in the course and rate of the silylation reactions, especially those employing ethoxysilane reagents. The NMR data are used to address the question of horizontal and vertical polymerization within the silane phase on the silica surface.

  1. Solid State NMR Investigations of Chain Dynamics and Network Order in Model Poly(dimethylsiloxane) Elastomers

    SciTech Connect

    Lewicki, J P; Mayer, B P; Wilson, T S; Chinn, S C; Maxwell, R S

    2010-12-09

    This work is at a relatively early stage, however it has been demonstrated that we can reliably probe basic network architectures using the MQ-NMR technique. The initial results are in good agreement with what is known from standard network theory and will serve as a basis for the study of progressively increasing structural complexity in Siloxane network systems.

  2. Polymorphs of Theophylline Characterized by DNP Enhanced Solid-State NMR.

    PubMed

    Pinon, Arthur C; Rossini, Aaron J; Widdifield, Cory M; Gajan, David; Emsley, Lyndon

    2015-11-01

    We show how dynamic nuclear polarization (DNP) enhanced solid-state NMR spectroscopy can be used to characterize polymorphs and solvates of organic solids. We applied DNP to three polymorphs and one hydrated form of the asthma drug molecule theophylline. For some forms of theophylline, sample grinding and impregnation with the radical-containing solution, which are necessary to prepare the samples for DNP, were found to induce polymorphic transitions or desolvation between some forms. We present protocols for sample preparation for solid-state magic-angle spinning (MAS) DNP experiments that avoid the polymorphic phase transitions in theophylline. These protocols include cryogrinding, grinding under inert atmosphere, and the appropriate choice of the impregnating liquid. By applying these procedures, we subsequently demonstrate that two-dimensional correlation experiments, such as (1)H-(13)C and (1)H-(15)N HETCOR or (13)C-(13)C INADEQUATE, can be obtained at natural isotopic abundance in reasonable times, thus enabling more advanced structural characterization of polymorphs. PMID:26393368

  3. Polymorphs of Theophylline Characterized by DNP Enhanced Solid-State NMR

    PubMed Central

    2015-01-01

    We show how dynamic nuclear polarization (DNP) enhanced solid-state NMR spectroscopy can be used to characterize polymorphs and solvates of organic solids. We applied DNP to three polymorphs and one hydrated form of the asthma drug molecule theophylline. For some forms of theophylline, sample grinding and impregnation with the radical-containing solution, which are necessary to prepare the samples for DNP, were found to induce polymorphic transitions or desolvation between some forms. We present protocols for sample preparation for solid-state magic-angle spinning (MAS) DNP experiments that avoid the polymorphic phase transitions in theophylline. These protocols include cryogrinding, grinding under inert atmosphere, and the appropriate choice of the impregnating liquid. By applying these procedures, we subsequently demonstrate that two-dimensional correlation experiments, such as 1H–13C and 1H–15N HETCOR or 13C–13C INADEQUATE, can be obtained at natural isotopic abundance in reasonable times, thus enabling more advanced structural characterization of polymorphs. PMID:26393368

  4. A solid-state NMR method to determine domain sizes in multi-component polymer formulations.

    PubMed

    Schlagnitweit, Judith; Tang, Mingxue; Baias, Maria; Richardson, Sara; Schantz, Staffan; Emsley, Lyndon

    2015-12-01

    Polymer domain sizes are related to many of the physical properties of polymers. Here we present a solid-state NMR experiment that is capable of measuring domain sizes in multi-component mixtures. The method combines selective excitation of carbon magnetization to isolate a specific component with proton spin diffusion to report on domain size. We demonstrate the method in the context of controlled release formulations, which represents one of today's challenges in pharmaceutical science. We show that we can measure domain sizes of interest in the different components of industrial pharmaceutical formulations at natural isotopic abundance containing various (modified) cellulose derivatives, such as microcrystalline cellulose matrixes that are film-coated with a mixture of ethyl cellulose (EC) and hydroxypropyl cellulose (HPC). PMID:26529201

  5. Proton decoupling and recoupling under double-nutation irradiation in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Takeda, Kazuyuki; Wakisaka, Asato; Takegoshi, K.

    2014-12-01

    The effect of 1H decoupling in magic-angle spinning solid-state NMR is studied under radiofrequency irradiation causing simultaneous nutations around a pair of orthogonal axes. Double-nutation with an arbitrary pair of nutation frequencies is implemented through modulation of the amplitude, phase, and frequency of the transmitting pulses. Similarity and difference of double-nutation decoupling and two-pulse phase-modulation decoupling schemes [A. E. Bennett, C. M. Rienstra, M. Auger, K. V. Lakshmi, and R. G. Griffin, J. Chem. Phys. 103, 6951-6958 (1995) and I. Scholz, P. Hodgkinson, B. H. Meier, and M. Ernst, J. Chem. Phys. 130, 114510 (2009)] are discussed. The structure of recoupling bands caused by interference of the 1H spin nutation with sample spinning is studied by both experiments and numerical simulations.

  6. A solid-state NMR method to determine domain sizes in multi-component polymer formulations

    NASA Astrophysics Data System (ADS)

    Schlagnitweit, Judith; Tang, Mingxue; Baias, Maria; Richardson, Sara; Schantz, Staffan; Emsley, Lyndon

    2015-12-01

    Polymer domain sizes are related to many of the physical properties of polymers. Here we present a solid-state NMR experiment that is capable of measuring domain sizes in multi-component mixtures. The method combines selective excitation of carbon magnetization to isolate a specific component with proton spin diffusion to report on domain size. We demonstrate the method in the context of controlled release formulations, which represents one of today's challenges in pharmaceutical science. We show that we can measure domain sizes of interest in the different components of industrial pharmaceutical formulations at natural isotopic abundance containing various (modified) cellulose derivatives, such as microcrystalline cellulose matrixes that are film-coated with a mixture of ethyl cellulose (EC) and hydroxypropyl cellulose (HPC).

  7. Characterisation of hydrogen bonding networks in RNAs via magic angle spinning solid state NMR spectroscopy.

    PubMed

    Riedel, Kerstin; Leppert, Jörg; Ohlenschläger, Oliver; Görlach, Matthias; Ramachandran, Ramadurai

    2005-04-01

    It is demonstrated that the spatial proximity of (1)H nuclei in hydrogen bonded base-pairs in RNAs can be conveniently mapped via magic angle spinning solid state NMR experiments involving proton spin diffusion driven chemical shift correlation of low gamma nuclei such as the imino and amino nitrogens of nucleic acid bases. As different canonical and non-canonical base-pairing schemes encountered in nucleic acids are characterised by topologically different networks of proton dipolar couplings, different base-pairing schemes lead to characteristic cross-peak intensity patterns in such correlation spectra. The method was employed in a study of a 100 kDa RNA composed of 97 CUG repeats, or (CUG)(97) that has been implicated in the neuromuscular disease myotonic dystrophy. (15)N-(15)N chemical shift correlation studies confirm the presence of Watson-Crick GC base pairs in (CUG)(97). PMID:15928999

  8. Intensity and mosaic spread analysis from PISEMA tensors in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Quine, J. R.; Achuthan, S.; Asbury, T.; Bertram, R.; Chapman, M. S.; Hu, J.; Cross, T. A.

    2006-04-01

    The solid-state NMR experiment PISEMA, is a technique for determining structures of proteins, especially membrane proteins, from oriented samples. One method for determining the structure is to find orientations of local molecular frames (peptide planes) with respect to the unit magnetic field direction, B0. This is done using equations that compute the coordinates of this vector in the frames. This requires an analysis of the PISEMA function and its degeneracies. As a measure of the sensitivity of peptide plane orientations to the data, we use these equations to derive a formula for the intensity function in the powder pattern. With this function and other measures, we investigate the effect of small changes in peptide plane orientations depending on the location of the resonances in the powder pattern spectrum. This gives us an indication of the change in lineshape due to mosaic spread and a way to interpret these in terms of an orientational error bar.

  9. Solid-state NMR study of geopolymer prepared by sol-gel chemistry

    SciTech Connect

    Tsai, Yi-Ling; Hanna, John V.; Lee, Yuan-Ling; Smith, Mark E.; Chan, Jerry C.C.

    2010-12-15

    Geopolymers are a new class of materials formed by the condensation of aluminosilicates and silicates obtained from natural minerals or industrial wastes. In this work, the sol-gel method is used to synthesize precursor materials for the preparation of geopolymers. The geopolymer samples prepared by our synthetic route have been characterized by a series of physical techniques, including Fourier-transform infrared, X-ray diffraction, and multinuclear solid-state NMR. The results are very similar to those obtained for the geopolymers prepared from natural kaolinite. We believe that our synthetic approach can offer a good opportunity for the medical applications of geopolymer. -- Graphical abstract: Geopolymer prepared by the sol-gel route has the same spectroscopic properties as the sample prepared from the natural kaolinite. Display Omitted

  10. High field 17O solid-state NMR study of alanine tripeptides

    NASA Astrophysics Data System (ADS)

    Yamauchi, Kazuo; Okonogi, Michi; Kurosu, Hiromichi; Tansho, Masataka; Shimizu, Tadashi; Gullion, Terry; Asakura, Tetsuo

    2008-02-01

    17O chemical shifts of Ala-Ala-Ala, with parallel and anti-parallel β-sheet structures, are observed using a 930-MHz high-resolution solid-state NMR spectrometer. Ala-Ala-Ala serves as a model sheet-forming peptide because it can be easily prepared as either a parallel or an anti-parallel β-sheet structure. Spectral differences between the two samples are observed which arise from molecular packing differences between the two sheet structures. DFT chemical shift calculations are performed for the two samples, and the calculated spectra are in good agreement with the experimental spectra. The DFT calculations provide insight into the nature of the chemical shift differences observed between the two sheet structures.

  11. Solid-state NMR studies of proteins immobilized on inorganic surfaces

    SciTech Connect

    Shaw, Wendy J.

    2015-09-01

    Solid state NMR is the primary tool for studying the quantitative, site-specific structure, orientation, and dynamics of biomineralization proteins under biologically relevant conditions. Two calcium phosphate proteins, statherin and leucine rich amelogenin protein (LRAP), have been studied in depth and have different features, challenging our ability to extract design principles. More recent studies of the significantly larger full-length amelogenin represent a challenging but necessary step to ultimately investigate the full diversity of biomineralization proteins. Interactions of amino acids and silaffin peptide with silica are also being studied, along with qualitative studies of proteins interacting with calcium carbonate. Dipolar recoupling techniques have formed the core of the quantitative studies, yet, the need for isolated spin pairs makes this approach costly and time intensive. The use of multi-dimensional techniques is advancing, methodology which, despite its challenges with these difficult-to-study proteins, will continue to drive future advancements in this area..

  12. Formation of Organic Molecular Nanocrystals under Rigid Confinement with Analysis by Solid State NMR.

    PubMed

    Yang, X; Ong, T C; Michaelis, V K; Heng, S; Huang, J; Griffin, R G; Myerson, A S

    2014-10-21

    Crystallization in rigid confinement is a promising method to obtain organic molecular nanocrystals. However, the crystallization behavior and the related characterization methods are not well studied. Here we present a systematic study of the nucleation of organic molecular nanocrystals in rigid pores. Four different compounds were studied, ibuprofen, fenofibrate, griseofulvin, and indomethacin, which range from simple to complex molecules. Solid-state Nuclear Magnetic Resonance (NMR) was employed to analyse the structure of these compounds inside pores which are difficult to characterize by other analytical methods. We successfully demonstrated the production of nano-crystalline ibuprofen, fenofibrate and griseofulvin in porous silica particles with ~ 40 nm pores. These nanocrystals showed significant enhancement in dissolution rates. These results help advance the fundamental understanding of nucleation under rigid confinement and may lead to potential applications in developing new formulations in the pharmaceutical industry. PMID:25258590

  13. Proton decoupling and recoupling under double-nutation irradiation in solid-state NMR

    SciTech Connect

    Takeda, Kazuyuki Wakisaka, Asato; Takegoshi, K.

    2014-12-14

    The effect of {sup 1}H decoupling in magic-angle spinning solid-state NMR is studied under radiofrequency irradiation causing simultaneous nutations around a pair of orthogonal axes. Double-nutation with an arbitrary pair of nutation frequencies is implemented through modulation of the amplitude, phase, and frequency of the transmitting pulses. Similarity and difference of double-nutation decoupling and two-pulse phase-modulation decoupling schemes [A. E. Bennett, C. M. Rienstra, M. Auger, K. V. Lakshmi, and R. G. Griffin, J. Chem. Phys. 103, 6951–6958 (1995) and I. Scholz, P. Hodgkinson, B. H. Meier, and M. Ernst, J. Chem. Phys. 130, 114510 (2009)] are discussed. The structure of recoupling bands caused by interference of the {sup 1}H spin nutation with sample spinning is studied by both experiments and numerical simulations.

  14. Solid-state NMR comparison of various spiders' dragline silk fiber.

    PubMed

    Creager, Melinda S; Jenkins, Janelle E; Thagard-Yeaman, Leigh A; Brooks, Amanda E; Jones, Justin A; Lewis, Randolph V; Holland, Gregory P; Yarger, Jeffery L

    2010-08-01

    Major ampullate (dragline) spider silk is a coveted biopolymer due to its combination of strength and extensibility. The dragline silk of different spiders have distinct mechanical properties that can be qualitatively correlated to the protein sequence. This study uses amino acid analysis and carbon-13 solid-state NMR to compare the molecular composition, structure, and dynamics of major ampullate dragline silk of four orb-web spider species ( Nephila clavipes , Araneus gemmoides , Argiope aurantia , and Argiope argentata ) and one cobweb species ( Latrodectus hesperus ). The mobility of the protein backbone and amino acid side chains in water exposed silk fibers is shown to correlate to the proline content. This implies that regions of major ampullate spidroin 2 protein, which is the only dragline silk protein with any significant proline content, become significantly hydrated in dragline spider silk. PMID:20593757

  15. NMR Determination of Protein pKa Values in the Solid State

    PubMed Central

    Schmidt, Heather L. Frericks; Shah, Gautam J.; Sperling, Lindsay J.; Rienstra, Chad M.

    2010-01-01

    Charged residues play an important role in defining key mechanistic features in many biomolecules. Determining the pKa values of large, membrane or fibrillar proteins can be challenging with traditional methods. In this study we show how solid-state NMR is used to monitor chemical shift changes during a pH titration for the small soluble β1 immunoglobulin binding domain of protein G. The chemical shifts of all the amino acids with charged side-chains throughout the uniformly-13C,15N-labeled protein were monitored over several samples varying in pH; pKa values were determined from these shifts for E27, D36, and E42, and the bounds for the pKa of other acidic side-chain resonances were determined. Additionally, this study shows how the calculated pKa values give insights into the crystal packing of the protein. PMID:20563223

  16. Formation of Organic Molecular Nanocrystals under Rigid Confinement with Analysis by Solid State NMR

    PubMed Central

    Yang, X.; Ong, T. C.; Michaelis, V. K.; Heng, S.; Huang, J.; Griffin, R. G.; Myerson, A. S.

    2014-01-01

    Crystallization in rigid confinement is a promising method to obtain organic molecular nanocrystals. However, the crystallization behavior and the related characterization methods are not well studied. Here we present a systematic study of the nucleation of organic molecular nanocrystals in rigid pores. Four different compounds were studied, ibuprofen, fenofibrate, griseofulvin, and indomethacin, which range from simple to complex molecules. Solid-state Nuclear Magnetic Resonance (NMR) was employed to analyse the structure of these compounds inside pores which are difficult to characterize by other analytical methods. We successfully demonstrated the production of nano-crystalline ibuprofen, fenofibrate and griseofulvin in porous silica particles with ~ 40 nm pores. These nanocrystals showed significant enhancement in dissolution rates. These results help advance the fundamental understanding of nucleation under rigid confinement and may lead to potential applications in developing new formulations in the pharmaceutical industry. PMID:25258590

  17. Characterization of the Vibrio cholerae Extracellular Matrix: A Top-Down Solid-State NMR Approach

    PubMed Central

    Reichhardt, Courtney; Fong, Jiunn C.N.; Yildiz, Fitnat; Cegelski, Lynette

    2015-01-01

    Bacterial biofilms are communities of bacterial cells surrounded by a self-secreted extracellular matrix. Biofilm formation by Vibrio cholerae, the human pathogen responsible for cholera, contributes to its environmental survival and infectivity. Important genetic and molecular requirements have been identified for V. cholerae biofilm formation, yet a compositional accounting of these parts in the intact biofilm or extracellular matrix has not been described. As insoluble and non-crystalline assemblies, determinations of biofilm composition pose a challenge to conventional biochemical and biophysical analysis. The V. cholerae extracellular matrix composition is particularly complex with several proteins, complex polysaccharides, and other biomolecules having been identified as matrix parts. We developed a new top-down solid-state NMR approach to spectroscopically assign and quantify the carbon pools of the intact V. cholerae extracellular matrix using 13C CPMAS and 13C{15N}, 15N{31P}, and 13C{31P}REDOR. General sugar, lipid, and amino acid pools were first profiled and then further annotated and quantified as specific carbon types, including carbonyls, amides, glycyl carbons, and anomerics. In addition, 15N profiling revealed a large amine pool relative to amide contributions, reflecting the prevalence of molecular modifications with free amine groups. Our top-down approach could be implemented immediately to examine the extracellular matrix from mutant strains that might alter polysaccharide production or lipid release beyond the cell surface; or to monitor changes that may accompany environmental variations and stressors such as altered nutrient composition, oxidative stress or antibiotics. More generally, our analysis has demonstrated that solid-state NMR is a valuable tool to characterize complex biofilm systems. PMID:24911407

  18. Specific inclusion mode of guest compounds in the amylose complex analyzed by solid state NMR spectroscopy.

    PubMed

    Tozuka, Yuichi; Takeshita, Aya; Nagae, Ayako; Wongmekiat, Arpansiree; Moribe, Kunikazu; Oguchi, Toshio; Yamamoto, Keiji

    2006-08-01

    The inclusion compound formation between linear amylose of molecular weight 102500 (AS100) and p-aminobenzoic acid (PA) during the sealed-heating process was investigated by powder X-ray diffractometry, infrared spectroscopy and solid state NMR spectroscopy. Sealed-heating of AS100 and PA at 100 degrees C for 6 h provided an inclusion compound with 6(1)-helix structure, while a 7(1)-helix structure was found when sealed-heating was carried out at 150 degrees C for 1 h. The formation of an inclusion compound was not observed when sealed-heating was performed at 50 degrees C for 6 h. The 7(1)-helix inclusion compound maintained its structure even during storage at high temperature while the 6(1)-helix inclusion compound decomposed and returned to the original V(a)-amylose upon heating to 180 degrees C. Quantitative determination revealed that one PA molecule could be included per one helical turn of AS100 for both 6(1)-helix and 7(1)-helix inclusion compounds. Solid state NMR spectroscopy suggested that PA molecules were included in the amylose helix core in the 7(1)-helix inclusion compound, while in the case of 6(1)-helix inclusion compound, PA molecules were accommodated in the interstices between amylose helices. Moreover, the inclusion compound formation by sealed-heating of AS100 was also observed when using PA analogues as guest compounds. The binding ratio of AS100 and PA analogues varied depending on the size of guest molecules. PMID:16880651

  19. Solid-State NMR Identification and Quantification of Newly Formed Aluminosilicate Phases in Weathered Kaolinite Systems

    SciTech Connect

    Crosson, Garry S.; Choi, Sunkyung; Chorover, Jon; Amistadi, Mary K.; O'Day, Peggy A.; Mueller, Karl T.

    2006-01-19

    The weathering of a specimen kaolinite clay was studied over the course of 369 d via solid-state 29Si magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy and high-field 27Al MAS NMR. The chosen baseline solution conditions (0.05 mol kg-1 of Al, 2 mol kg-1 of Na+, 1 mol kg-1 of NO3 -, 1 mol kg-1 of OH-, and pH ~13.8) approximate those of solutions leaking from waste tanks at the Hanford Site in Richland, WA. Nonradioactive Cs and Sr cations were added to this synthetic tank waste leachate (STWL) solution at concentrations of 10-3, 10-4, and 10-5 molal (m) to represent their radionuclide counterparts. The transformations of silicon- and aluminum-containing solid phase species were monitored quantitatively by using NMR spectroscopy, with the resulting spectra directly reporting the influence of the initial Cs and Sr on formation and transformation of the neo-formed solids. At the lowest concentration of Cs and Sr employed (10-5 m in each cation) peaks consistent with the formation of zeolite-like minerals were detected via 29Si and 27Al MAS NMR as early as 33 d. At concentrations of 10-3 m in each cation, new silicon species are not detected until 93 d, although neophases containing four-coordinate aluminum were detectable at earlier reaction times via 27Al MAS NMR. At the highest magnetic field strengths employed in this NMR study, deconvolutions of resonances detected in the tetrahedral region of the 27Al MAS spectra yielded multiple components, indicating the existence of at least four new aluminum-containing phases. Two of these phases are identified as sodalite and cancrinite through comparison with diffuse-reflectance infrared (DRIFT) spectra and powder X-ray diffraction (XRD) results, while a third phase may correlate with a previously detected aluminum-rich chabazite phase. All measurable solid reaction products have been quantified via their 27Al MAS resonances acquired at high magnetic field strengths (17.6 T), and the quantitative nature of the 27Al NMR data shows that cancrinite growth increases while sodalite reaches a steady state with respect to total aluminum in the solid phases. The data also relate the coupling of Cs sorption to the ripening of feldspathoid phases in this heterogeneous system as a function of time, and illustrate the important influence of co-contaminants on the environmental reaction kinetics studied here.

  20. SIMPSON: A general simulation program for solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Bak, Mads; Rasmussen, Jimmy T.; Nielsen, Niels Chr.

    2011-12-01

    A computer program for fast and accurate numerical simulation of solid-state NMR experiments is described. The program is designed to emulate a NMR spectrometer by letting the user specify high-level NMR concepts such as spin systems, nuclear spin interactions, RF irradiation, free precession, phase cycling, coherence-order filtering, and implicit/explicit acquisition. These elements are implemented using the Tel scripting language to ensure a minimum of programming overhead and direct interpretation without the need for compilation, while maintaining the flexibility of a full-featured programming language. Basicly, there are no intrinsic limitations to the number of spins, types of interactions, sample conditions (static or spinning, powders, uniaxially oriented molecules, single crystals, or solutions), and the complexity or number of spectral dimensions for the pulse sequence. The applicability ranges from simple ID experiments to advanced multiple-pulse and multiple-dimensional experiments, series of simulations, parameter scans, complex data manipulation/visualization, and iterative fitting of simulated to experimental spectra. A major effort has been devoted to optimizing the computation speed using state-of-the-art algorithms for the time-consuming parts of the calculations implemented in the core of the program using the C programming language. Modification and maintenance of the program are facilitated by releasing the program as open source software (General Public License) currently at http://nmr.imsb.au.dk. The general features of the program are demonstrated by numerical simulations of various aspects for REDOR, rotational resonance, DRAMA, DRAWS, HORROR, C7, TEDOR, POST-C7, CW decoupling, TPPM, F-SLG, SLF, SEMA-CP, PISEMA, RFDR, QCPMG-MAS, and MQ-MAS experiments.

  1. SIMPSON: A General Simulation Program for Solid-State NMR Spectroscopy

    NASA Astrophysics Data System (ADS)

    Bak, Mads; Rasmussen, Jimmy T.; Nielsen, Niels Chr.

    2000-12-01

    A computer program for fast and accurate numerical simulation of solid-state NMR experiments is described. The program is designed to emulate a NMR spectrometer by letting the user specify high-level NMR concepts such as spin systems, nuclear spin interactions, RF irradiation, free precession, phase cycling, coherence-order filtering, and implicit/explicit acquisition. These elements are implemented using the Tcl scripting language to ensure a minimum of programming overhead and direct interpretation without the need for compilation, while maintaining the flexibility of a full-featured programming language. Basicly, there are no intrinsic limitations to the number of spins, types of interactions, sample conditions (static or spinning, powders, uniaxially oriented molecules, single crystals, or solutions), and the complexity or number of spectral dimensions for the pulse sequence. The applicability ranges from simple 1D experiments to advanced multiple-pulse and multiple-dimensional experiments, series of simulations, parameter scans, complex data manipulation/visualization, and iterative fitting of simulated to experimental spectra. A major effort has been devoted to optimizing the computation speed using state-of-the-art algorithms for the time-consuming parts of the calculations implemented in the core of the program using the C programming language. Modification and maintenance of the program are facilitated by releasing the program as open source software (General Public License) currently at http://nmr.imsb.au.dk. The general features of the program are demonstrated by numerical simulations of various aspects for REDOR, rotational resonance, DRAMA, DRAWS, HORROR, C7, TEDOR, POST-C7, CW decoupling, TPPM, F-SLG, SLF, SEMA-CP, PISEMA, RFDR, QCPMG-MAS, and MQ-MAS experiments.

  2. Quantitative solid-state 13C NMR with signal enhancement by multiple cross polarization

    NASA Astrophysics Data System (ADS)

    Johnson, Robert L.; Schmidt-Rohr, Klaus

    2014-02-01

    A simple new method is presented that yields quantitative solid-state magic-angle spinning (MAS) 13C NMR spectra of organic materials with good signal-to-noise ratios. It achieves long (>10 ms) cross polarization (CP) from 1H without significant magnetization losses due to relaxation and with a moderate duty cycle of the radio-frequency irradiation, by multiple 1-ms CP periods alternating with 1H spin-lattice relaxation periods that repolarize the protons. The new method incorporates previous techniques that yield less distorted CP/MAS spectra, such as a linear variation (“ramp”) of the radio-frequency field strength, and it overcomes their main limitation, which is T1ρ relaxation of the spin-locked 1H magnetization. The ramp of the radio-frequency field strength and the asymptotic limit of cross polarization makes the spectral intensity quite insensitive to the exact field strengths used. The new multiCP pulse sequence is a “drop-in” replacement for previous CP methods and produces no additional data-processing burden. Compared to the only reliable quantitative 13C NMR method for unlabeled solids previously available, namely direct-polarization NMR, the measuring time is reduced by more than a factor of 50, enabling higher-throughput quantitative NMR studies. The new multiCP technique is validated with 14-kHz MAS on amino-acid derivatives, plant matter, a highly aromatic humic acid, and carbon materials made by low-temperature pyrolysis.

  3. Solid state NMR method development and studies of biological and biomimetic nanocomposites

    SciTech Connect

    Hu, Yanyan

    2011-02-07

    This thesis describes application and development of advanced solid-state nuclear magnetic resonance techniques for complex materials, in particular organic-inorganic nanocomposites and thermoelectric tellurides. The apatite-collagen interface, essential for understanding the biomineralization process in bone and engineering the interface for controlled bio-mimetic synthesis and optimized mechanical properties, is buried within the nanocomposite of bone. We used multinuclear solid-state NMR to study the composition and structure of the interface. Citrate has been identified as the main organic molecule strongly bound to the apatite surface with a density of 1/(2 nm){sup 2}, covering 1/6 of the total surface area in bovine bone. Citrate provides more carboxylate groups, one of the key functional groups found to affect apatite nucleation and growth, than all the non-collagenous proteins all together in bone; thus we propose that citrate stabilizes apatite crystals at a very small thickness of {approx}3 nm (4 unit cells) to increase bone fracture tolerance. The hypothesis has been confirmed in vitro by adding citrate in the bio-mimetic synthesis of polymerhydroxyapatite nanocomposites. The results have shown that the size of hydroxyapatite nanocrystals decreases as increasing citrate concentration. With citrate concentrations comparable to that in body fluids, similar-sized nanocrystals as in bone have been produced. Besides the dimensions of the apatite crystals, the composition of bone also affects its biofunctional and macroscopic mechanical properties; therefore, our team also extended its effort to enhance the inorganic portion in our bio-mimetic synthesis from originally 15 wt% to current 50 wt% compared to 65 wt% in bovine bone, by using Lysine-Leucine hydroxyapatite nucleating diblock co-polypeptide, which forms a gel at very low concentration. In this thesis, various advanced solid state NMR techniques have been employed to characterize nanocomposites. Meanwhile, we have developed new methods to achieve broadband high resolution NMR and improve the accuracy of inter-nuclear distance measurements involving quadrupolar spins. Broadband high resolution NMR of spin-1/2 nuclei has been accomplished by the adaptation of the magic angle turning (MAT) method to fast magic angle spinning, termed fast MAT, by solving technical problems such as off resonance effects. Fast MAT separates chemical shift anisotropy and isotropic chemical shifts over a spectral range of {approx}1.8 {gamma}B{sub 1} without significant distortions. Fast MAT {sup 125}Te NMR has been applied to study technologically important telluride materials with spectra spreading up to 190 kHz. The signal-to-noise ratio of the spectra is significantly improved by using echo-matched Gaussian filtering in offline data processing. The accuracy of the measured distances between spin-1/2 and quadrupolar nuclei with methods such as SPIDER and REAPDOR has been improved by compensating for the fast longitudinal quadrupolar relaxation on the sub-millisecond with a modified S{sub 0} pulse sequence. Also, the T1Q effect on the spin coherence and its spinning speed dependency has been explored and documented with analytical and numerical simulations as well as experimental measurements.

  4. Preparation of uniformly isotope labeled KcsA for solid state NMR: expression, purification, reconstitution into liposomes and functional assay.

    PubMed

    Bhate, Manasi P; Wylie, Benjamin J; Thompson, Ameer; Tian, Lin; Nimigean, Crina; McDermott, Ann E

    2013-10-01

    We report the expression, purification, liposome reconstitution and functional validation of uniformly (13)C and (15)N isotope labeled KcsA, a bacterial potassium channel that has high homology with mammalian channels, for solid-state NMR studies. The expression and purification is optimized for an average yield of ∼35-40mg/L of M9 media in a time-efficient way. The protein purity is confirmed by gel electrophoresis and the protein concentration is quantified by UV-vis absorption spectroscopy. Protocols to efficiently reconstitute KcsA into liposomes are also presented. The presence of liposomes is confirmed by cryo-electron microscopy images and the effect of magic angle spinning on liposome packing is shown. High-resolution solid-state NMR spectra of uniformly isotope labeled KcsA in these liposomes reveal that our protocol yields to a very homogenous KcsA sample with high signal to noise and several well-resolved residues in NMR spectra. Electrophysiology of our samples before and after solid-state NMR show that channel function and selectivity remain intact after the solid-state NMR. PMID:23916531

  5. Preparation of uniformly isotope labeled KcsA for solid state NMR: Expression, purification, reconstitution into liposomes and functional assay

    PubMed Central

    Bhate, Manasi P.; Wylie, Benjamin J.; Thompson, Ameer; Tian, Lin; Nimigean, Crina; McDermott, Ann E.

    2013-01-01

    We report the expression, purification, liposome reconstitution and functional validation of uniformly 13C and 15N isotope labeled KcsA, a bacterial potassium channel that has high homology with mammalian channels, for solid-state NMR studies. The expression and purification is optimized for an average yield of ~ 35–40 milligrams per liter of M9 media in a time-efficient way. The protein purity is confirmed by gel electrophoresis and the protein concentration is quantified by UV-Vis absorption spectroscopy. Protocols to efficiently reconstitute KcsA into liposomes are also presented. The presence of liposomes is confirmed by cryo-electron microscopy images and the effect of magic angle spinning on liposome packing is shown. High-resolution solid-state NMR spectra of uniformly isotope labeled KcsA in these liposomes reveal that our protocol yields to a very homogenous KcsA sample with high signal to noise and several well-resolved residues in NMR spectra. Electrophysiology of our samples before and after solid-state NMR show that channel function and selectivity remain intact after the solid-state NMR. PMID:23916531

  6. Structural Analysis of Nanoscale Self-Assembled Discoidal Lipid Bilayers by Solid-State NMR Spectroscopy

    PubMed Central

    Li, Ying; Kijac, Aleksandra Z.; Sligar, Stephen G.; Rienstra, Chad M.

    2006-01-01

    Nanodiscs are an example of discoidal nanoscale self-assembled lipid/protein particles similar to nascent high-density lipoproteins, which reduce the risk of coronary artery disease. The major protein component of high-density lipoproteins is human apolipoprotein A-I, and the corresponding protein component of Nanodiscs is membrane scaffold protein 1 (MSP1), a 200-residue lipid-binding domain of human apolipoprotein A-I. Here we present magic-angle spinning (MAS) solid-state NMR studies of uniformly 13C,15N-labeled MSP1 in polyethylene glycol precipitated Nanodiscs. Two-dimensional MAS 13C-13C correlation spectra show excellent microscopic order of MSP1 in precipitated Nanodiscs. Secondary isotropic chemical shifts throughout the protein are consistent with a predominantly helical structure. Moreover, the backbone conformations of prolines derived from their 13C chemical shifts are consistent with the molecular belt model but not the picket fence model of lipid-bound MSP1. Overall comparison of experimental spectra and 13C chemical shifts predicted from several structural models also favors the belt model. Our study thus supports the belt model of Nanodisc structure and demonstrates the utility of MAS NMR to study the structure of high molecular weight lipid-protein complexes. PMID:16905610

  7. 13C and 15N solid state NMR of piezoelectric nylons

    NASA Astrophysics Data System (ADS)

    Mathias, Lon J.; Johnson, C. G.; Steadman, Scott J.

    1993-07-01

    Solid state nuclear magnetic resonance (NMR) has been shown to be a unique spectroscopic tool for determining molecular composition, crystallinity, packing, orientation and motion in as-obtained and end-use materials. We have developed several methods for evaluating the molecular-level properties and behavior of polymeric materials, especially piezoelectric nylons. Analysis of nylon 7 homopolymer under various sample treatment histories related to poling and generation of piezoelectric properties allows qualitative evaluation of the two main types of crystal forms present, the (alpha) -form which appears to be the one responsible for piezoelectric behavior in this polymer, and the (gamma) -form which can co-exist with the (alpha) -form in some samples. Based on the possibility that molecular composition could be used to control crystallinity and microscopic packing, and thereby affect macroscopic properties such as piezoelectricity, we have synthesized and characterized two families of nylon co-polymers consisting of even-odd A-B monomer combinations. We have determined degrees and types of crystallinity for these materials using a combination of thermal, FTIR and NMR, measurements. The molecular-level behavior of these materials is related to observed properties. Evaluation of piezoelectric properties is underway, and initial results are summarized.

  8. Atomic-resolution structure of cytoskeletal bactofilin by solid-state NMR*

    PubMed Central

    Shi, Chaowei; Fricke, Pascal; Lin, Lin; Chevelkov, Veniamin; Wegstroth, Melanie; Giller, Karin; Becker, Stefan; Thanbichler, Martin; Lange, Adam

    2015-01-01

    Bactofilins are a recently discovered class of cytoskeletal proteins of which no atomic-resolution structure has been reported thus far. The bacterial cytoskeleton plays an essential role in a wide range of processes, including morphogenesis, cell division, and motility. Among the cytoskeletal proteins, the bactofilins are bacteria-specific and do not have a eukaryotic counterpart. The bactofilin BacA of the species Caulobacter crescentus is not amenable to study by x-ray crystallography or solution nuclear magnetic resonance (NMR) because of its inherent noncrystallinity and insolubility. We present the atomic structure of BacA calculated from solid-state NMR–derived distance restraints. We show that the core domain of BacA forms a right-handed β helix with six windings and a triangular hydrophobic core. The BacA structure was determined to 1.0 Å precision (heavy-atom root mean square deviation) on the basis of unambiguous restraints derived from four-dimensional (4D) HN-HN and 2D C-C NMR spectra. PMID:26665178

  9. Comparison among Magnus/Floquet/Fer expansion schemes in solid-state NMR

    SciTech Connect

    Takegoshi, K. Miyazawa, Norihiro; Sharma, Kshama; Madhu, P. K.

    2015-04-07

    We here revisit expansion schemes used in nuclear magnetic resonance (NMR) for the calculation of effective Hamiltonians and propagators, namely, Magnus, Floquet, and Fer expansions. While all the expansion schemes are powerful methods there are subtle differences among them. To understand the differences, we performed explicit calculation for heteronuclear dipolar decoupling, cross-polarization, and rotary-resonance experiments in solid-state NMR. As the propagator from the Fer expansion takes the form of a product of sub-propagators, it enables us to appreciate effects of time-evolution under Hamiltonians with different orders separately. While 0th-order average Hamiltonian is the same for the three expansion schemes with the three cases examined, there is a case that the 2nd-order term for the Magnus/Floquet expansion is different from that obtained with the Fer expansion. The difference arises due to the separation of the 0th-order term in the Fer expansion. The separation enables us to appreciate time-evolution under the 0th-order average Hamiltonian, however, for that purpose, we use a so-called left-running Fer expansion. Comparison between the left-running Fer expansion and the Magnus expansion indicates that the sign of the odd orders in Magnus may better be reversed if one would like to consider its effect in order.

  10. Comparison among Magnus/Floquet/Fer expansion schemes in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Takegoshi, K.; Miyazawa, Norihiro; Sharma, Kshama; Madhu, P. K.

    2015-04-01

    We here revisit expansion schemes used in nuclear magnetic resonance (NMR) for the calculation of effective Hamiltonians and propagators, namely, Magnus, Floquet, and Fer expansions. While all the expansion schemes are powerful methods there are subtle differences among them. To understand the differences, we performed explicit calculation for heteronuclear dipolar decoupling, cross-polarization, and rotary-resonance experiments in solid-state NMR. As the propagator from the Fer expansion takes the form of a product of sub-propagators, it enables us to appreciate effects of time-evolution under Hamiltonians with different orders separately. While 0th-order average Hamiltonian is the same for the three expansion schemes with the three cases examined, there is a case that the 2nd-order term for the Magnus/Floquet expansion is different from that obtained with the Fer expansion. The difference arises due to the separation of the 0th-order term in the Fer expansion. The separation enables us to appreciate time-evolution under the 0th-order average Hamiltonian, however, for that purpose, we use a so-called left-running Fer expansion. Comparison between the left-running Fer expansion and the Magnus expansion indicates that the sign of the odd orders in Magnus may better be reversed if one would like to consider its effect in order.

  11. Optimizing oriented planar-supported lipid samples for solid-state protein NMR.

    PubMed

    Rainey, Jan K; Sykes, Brian D

    2005-10-01

    Sample orientation relative to the static magnetic field of an NMR spectrometer allows study of membrane proteins in the lipid bilayer setting. The straightforward preparation and handling of extremely thin mica substrates with consistent surface properties has prompted us to examine oriented phospholipid bilayer and hexagonal phases on mica. The spectral characteristics of oriented lipid samples formed on mica are as good as or better than those on glass. Nine solvents with varying dielectric constants were used to cast lipid films or for vesicle spreading; film characteristics were then compared, and static solid-state 31P-NMR was used to characterize the degree of orientation of the hydrated lipid species. Lipids with four headgroup chemistries were tested: 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), 1,2-dioleoyl-sn-glycero-3-phosphate (DOPA), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). Solvent affected orientation of POPG, DOPA, and DOPE, but not POPC. Film characteristics varied with solvent, with ramifications for producing homogeneous oriented lipid samples. POPC was used to optimize the amount of lipid per substrate and compare hydration methods. POPG did not orient reproducibly, whereas POPG-POPC mixtures did. DOPA showed 1-2 oriented states depending upon hydration level and deposition method. DOPE formed an oriented hexagonal phase that underwent a reversible temperature-induced phase transition to the oriented bilayer phase. PMID:16085766

  12. Frequency-swept pulse sequences for 19F heteronuclear spin decoupling in solid-state NMR.

    PubMed

    Chandran, C Vinod; Madhu, P K; Wormald, Philip; Bräuniger, Thomas

    2010-10-01

    Heteronuclear spin decoupling pulse sequences in solid-state NMR have mostly been designed and applied for irradiating 1H as the abundant nucleus. Here, a systematic comparison of different methods for decoupling 19F in rigid organic solids is presented, with a special emphasis on the recently introduced frequency-swept sequences. An extensive series of NMR experiments at different MAS frequencies was conducted on fluorinated model compounds, in combination with large sets of numerical simulations. From both experiments and simulations it can be concluded that the frequency-swept sequences SWf-TPPM and SWf-SPINAL deliver better and more robust spin decoupling than the original sequences SPINAL and TPPM. Whereas the existence of a large chemical shift anisotropy and isotropic shift dispersion for 19F does compromise the decoupling efficiency, the relative performance hierarchy of the sequences remains unaffected. Therefore, in the context of rigid organic solids under moderate MAS frequencies, the performance trends observed for 19F decoupling are very similar to those observed for 1H decoupling. PMID:20729111

  13. Solid-state NMR characterization of tri-ethyleneglycol grafted polyisocyanopeptides.

    PubMed

    Zinkevich, T; Venderbosch, B; Jaspers, M; Kouwer, P H J; Rowan, A E; van Eck, E R H; Kentgens, A P M

    2016-04-01

    In aqueous media, ethylene glycol substituted polyisocyanopeptides (PICPs) change their state (undergo a sol-to-gel transition) as a response to temperature. This makes them promising materials for various biomedical applications, for instance, for controlled drug release and non-damaging wound dressing. To utilize PICP in biomedical applications, understanding of the origin of the gelation process is needed, but this is experimentally difficult because of the notoriously low gelator concentration in combination with the slow polymer dynamics in the sample. This paper describes a detailed characterization of the dried state of PICPs by solid-state NMR measurements. Both the (13) C and the (1) H NMR resonances were assigned using a combination of 1D cross-polarization magic angle spinning, 2D (13) C-(1) H heteronuclear correlation spectra and (1) H-(1) H single quantum-double quantum experiments. In addition, the chemical groups involved in dipolar interaction with each other were used to discuss the dynamics and spatial conformation of the polymer. In contrast to other PICP polymers, two resonances for the backbone carbon are observed, which are present in equal amounts. The possible origin of these resonances is discussed in the last section of this work. The data obtained during the current studies will be further used in elucidating mechanisms of the bundling and gelation. A comprehensive picture will make it possible to tailor polymer properties to meet specific needs in different applications. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26559660

  14. Solid-State NMR Evidence for β-Hairpin Structure within MAX8 Designer Peptide Nanofibers

    PubMed Central

    Leonard, Sarah R.; Cormier, Ashley R.; Pang, Xiaodong; Zimmerman, Maxwell I.; Zhou, Huan-Xiang; Paravastu, Anant K.

    2013-01-01

    MAX8, a designer peptide known to undergo self-assembly following changes in temperature, pH, and ionic strength, has demonstrated usefulness for tissue engineering and drug delivery. It is hypothesized that the self-assembled MAX8 nanofiber structure consists of closed β-hairpins aligned into antiparallel β-sheets. Here, we report evidence from solid-state NMR spectroscopy that supports the presence of the hypothesized β-hairpin conformation within the nanofiber structure. Specifically, our 13C-13C two-dimensional exchange data indicate spatial proximity between V3 and K17, and 13C-13C dipolar coupling measurements reveal proximity between the V3 and V18 backbone carbonyls. Moreover, isotopic dilution of labeled MAX8 nanofibers did not result in a loss of the 13C-13C dipolar couplings, showing that these couplings are primarily intramolecular. NMR spectra also indicate the existence of a minor conformation, which is discussed in terms of previously hypothesized nanofiber physical cross-linking and possible nanofiber polymorphism. PMID:23823242

  15. Sensitivity-enhanced solid-state NMR detection of expansin's target in plant cell walls

    SciTech Connect

    Wang, Tuo; Park, Yong Bum; Caporini, Marc A.; Rosay, Melanie; Zhong, Linghao; Cosgrove, Daniel J.; Hong, Mei

    2013-08-29

    Structure determination of protein binding to noncrystalline macromolecular assemblies such as plant cell walls (CWs) poses a significant structural biology challenge. CWs are loosened during growth by expansin proteins, which weaken the noncovalent network formed by cellulose, hemicellulose, and pectins, but the CW target of expansins has remained elusive because of the minute amount of the protein required for activity and the complex nature of the CW. Using solid-state NMR spectroscopy, combined with sensitivity-enhancing dynamic nuclear polarization (DNP) and differential isotopic labeling of expansin and polysaccharides, we have now determined the functional binding target of expansin in the Arabidopsis thaliana CW. By transferring the electron polarization of a biradical dopant to the nuclei, DNP allowed selective detection of 13C spin diffusion from trace concentrations of 13C, 15N-labeled expansin in the CW to nearby polysaccharides. From the spin diffusion data of wild-type and mutant expansins, we conclude that to loosen the CW, expansin binds highly specific cellulose domains enriched in xyloglucan, whereas more abundant binding to pectins is unrelated to activity. Molecular dynamics simulations indicate short 13C-13C distances of 46 between a hydrophobic surface of the cellulose microfibril and an aromatic motif on the expansin surface, consistent with the observed NMR signals. DNP-enhanced 2D 13C correlation spectra further reveal that the expansin-bound cellulose has altered conformation and is enriched in xyloglucan, thus providing unique insight into the mechanism of CW loosening. DNP-enhanced NMR provides a powerful, generalizable approach for investigating protein binding to complex macromolecular targets.

  16. Sensitivity-enhanced solid-state NMR detection of expansin's target in plant cell walls

    SciTech Connect

    Wang, Tuo; Park, Yong Bum; Caporini, Marc A.; Rosay, Melanie; Zhong, Linghao; Cosgrove, Daniel J.; Hong, Mei

    2013-08-29

    Structure determination of protein binding to noncrystalline macromolecular assemblies such as plant cell walls (CWs) poses a significant structural biology challenge. CWs are loosened during growth by expansin proteins, which weaken the noncovalent network formed by cellulose, hemicellulose, and pectins, but the CW target of expansins has remained elusive because of the minute amount of the protein required for activity and the complex nature of the CW. Using solid-state NMR spectroscopy, combined with sensitivity-enhancing dynamic nuclear polarization (DNP) and differential isotopic labeling of expansin and polysaccharides, we have now determined the functional binding target of expansin in the Arabidopsis thaliana CW. By transferring the electron polarization of a biradical dopant to the nuclei, DNP allowed selective detection of 13C spin diffusion from trace concentrations of 13C, 15N-labeled expansin in the CW to nearby polysaccharides. From the spin diffusion data of wild-type and mutant expansins, we conclude that to loosen the CW, expansin binds highly specific cellulose domains enriched in xyloglucan, whereas more abundant binding to pectins is unrelated to activity. Molecular dynamics simulations indicate short 13C-13C distances of 4–6 Å between a hydrophobic surface of the cellulose microfibril and an aromatic motif on the expansin surface, consistent with the observed NMR signals. DNP-enhanced 2D 13C correlation spectra further reveal that the expansin-bound cellulose has altered conformation and is enriched in xyloglucan, thus providing unique insight into the mechanism of CW loosening. DNP-enhanced NMR provides a powerful, generalizable approach for investigating protein binding to complex macromolecular targets.

  17. Soil organic matter dynamics as characterized with 1H and 13C solid-state NMR techniques

    NASA Astrophysics Data System (ADS)

    Jäger, Alex; Schwarz, Jette; Bertmer, Marko; Schaumann, Gabriele E.

    2010-05-01

    Soil organic matter (SOM) is a complex and heterogeneous matter. Characterization by solid-state NMR methods on 1H and 13C nuclei is therefore demanding. Our goal is to obtain information on the dynamic behaviour of soil samples and to study the influence of external parameters on both structure and dynamics. We regard water molecules to be the pivotal agent of soil dynamics by generating a network between organic matter via intermolecular hydrogen bonding, which leads to cross linking of organic matter and increases its rigidity. Although 1H solid-state NMR on non-rotating samples are not so commonly used for soil characterization, they enable the differentiation of proton mobilities via their linewidths which are resulting from differences in the dipole-dipole coupling strengths. Therefore, even weak molecular interactions such as hydrogen bonding can be differentiated and changes due to heat treatments and the short and long term behaviour followed. Though in principle a simple technique, static 1H measurements are complicated by several means, one of them is the high abundance in almost all matter including probe head material that has to be excluded for analysis. Finally, we selected 1H DEPTH [1] and Hahn-echo sequences to distinguish different mobilities in soil, mainly free moving water and water fixed in the soil matrix. After decomposition using Gaussian and Lorentzian lineshapes, the relative amounts of mobile and rigid water molecules can be obtained. By heating the samples above 100°C in sealed glass tubes, the proposed water network is destroyed and able to rebuild after cooling. This long term behaviour is studied on the course of months. Furthermore, the instant changes before and after heating are shown for a series of soil samples to characterize soils based on this water network model. To combine the information obtained on the 1H mobility with focus on water dynamics, 13C 2D WISE (wideline separation) measurements were done. This method yields 1H mobilities of carbon containing molecules, in our case the soil organic matter. On the one hand, this can be correlated with the results from the static 1H measurements and on the other hand, mobility changes before, during and after a heating event can be studied. Combining the various information from NMR together with data from DSC (differential scanning calorimetry), a better understanding and perhaps a contribution to a modern model of soil dynamics can be reached.

  18. Structure and Transformation of Amorphous Calcium Carbonate: A Solid-State 43Ca NMR and Computational Molecular Dynamics Investigation

    SciTech Connect

    Singer, Jared W.; Yazaydin, A. O.; Kirkpatrick, Robert J.; Bowers, Geoffrey M.

    2012-05-22

    Amorphous calcium carbonate (ACC) is a metastable precursor to crystalline CaCO{sub 3} phases that precipitates by aggregation of ion pairs and prenucleation clusters. We use {sup 43}Ca solid-state NMR spectroscopy to probe the local structure and transformation of ACC synthesized from seawater-like solutions with and without Mg{sup 2+} and computational molecular dynamics (MD) simulations to provide more detailed molecular-scale understanding of the ACC structure. The {sup 43}Ca NMR spectra of ACC collected immediately after synthesis consist of broad, featureless resonances with Gaussian line shapes (FWHH = 27.6 {+-} 1 ppm) that do not depend on Mg{sup 2+} or H{sub 2}O content. A correlation between {sup 43}Ca isotropic chemical shifts and mean Ca-O bond distances for crystalline hydrous and anhydrous calcium carbonate phases indicates indistinguishable maximum mean Ca-O bond lengths of {approx}2.45 {angstrom} for all our samples. This value is near the upper end of the published Ca-O bond distance range for biogenic and synthetic ACCs obtained by Ca-X-ray absorption spectroscopy. It is slightly smaller than the values from the structural model of Mgfree ACC by Goodwin et al. obtained from reverse Monte Carlo (RMC) modeling of X-ray scattering data and our own computational molecular dynamics (MD) simulation based on this model. An MD simulation starting with the atomic positions of the Goodwin et al. RMC model using the force field of Raiteri and Gale shows significant structural reorganization during the simulation and that the interconnected carbonate/water-rich channels in the Goodwin et al. model shrink in size over the 2 ns simulation time. The distribution of polyhedrally averaged Ca-O bond distances from the MD simulation is in good agreement with the {sup 43}Ca NMR peak shape, suggesting that local structural disorder dominates the experimental line width of ACC.

  19. Investigation of Local Structures in Layered Niobates by Solid-state NMR Spectroscopy

    NASA Astrophysics Data System (ADS)

    Liu, Ting

    Research on ion-exchangeable layered niobates has attracted great attention due to their unique structures and corresponding variations in properties and applications, such as ion conductors, solid acids, and water splitting catalysts. Families of layered niobates include double-layered or triple-layered Dion-Jacobson type perovskites (ALaNb2O7, A = Cs, Rb, K, H; AM2Nb3O10, A = Rb, K, H; M = Sr, Ca), layered niobates with both edge and corner sharing of NbO6 octahedra (KNb3O8, HNb3O6, Nb 6O17 and H4Nb6O17) and many others. Lately, more developments in the layered niobates through a variety of topochemical manipulations have been achieved. The topochemical reactions include ion exchange, exfoliation, substitution, and etc. As a result, many new materials have been successfully prepared, for example, solid solutions (ALa2NbTi2O10, ACaLaNb2TiO 10 and ACa2Nb3-xTaxO10, etc.), nanosheets (HNb3O8, H4Nb6O17, HLaNb2O7, HCa2Nb3O10, etc., to intercalate with organic molecules such as tetrabutylammonium hydroxide or n-butylamines), and nanoscrolls (from H2K2Nb 6O17). While these structural modifications often induce improvements in properties, the fundamental mechanisms of improvements in properties upon the modifications, especially local structural arrangements are poorly understood, which is often limited by structural characterizations. Particularly, the characterizations of the exfoliated nanosheets can be difficult by conventional X-ray diffraction (XRD) method due to disordered structures. Alternatively, solid-state nuclear magnetic resonance (NMR) spectroscopy is a useful tool to study local structures in solids. The structural information can be extracted by examining intrinsic interactions, such as quadrupolar, chemical shielding, and dipolar interactions, which are all associated with local environments surrounding a specific nucleus, 1H or 93Nb in layered niobates. The ultimate goal of this dissertation is to understand the relationships between local structures of layered niobates and their chemical or physical properties, and provide insights into further modifications and improvements. The primary objectives of this work are summarized below: I. Synthesis of series of layered niobates (ALaNb2O7 , A = Cs, Rb, K; KNb3O8; K4Nb 6O17; RbLa2NbTi2O10 and RbCaLaNb2TiO10) by microwave heating or cation exchange methods, their protonated forms by acid exchange (HLaNb2O 7, H3ONb3O8 and HNb3O 8, H4Nb8O17, HLa2NbTi 2O10 and HCaLaNb2TiO10), and three nanosheet niobates by exfoliation (HNb3O8, H4Nb 6O17 and HLaNb2O7 nanosheets). II. Structural characterizations of all niobates by powder XRD and solid-state NMR spectroscopy. Powder XRD is used to determine lattice constants and long-range structural ordering. Solid-state NMR is used to determine the electric field gradient parameters, chemical shift anisotropy parameters and dipolar coupling constants. Solid-state NMR techniques include 93Nb MQMAS, wide-line VOCS echo and WURST-echo; 1H{93Nb} CP, TRAPDOR, S-RESPDOR and iS-RESPDOR experiments. III. Understanding the trends of changes in NMR parameters with respect to cation exchange, exfoliation and compositional alteration, and correlation of the NMR parameters with local environments and possible structural rearrangements. IV. Identification of proton locations in the acid-exchanged niobates and surface acidity for the exfoliated nanosheets, based on 1H chemical shifts and dipolar coupling information from CP, S-RESPDOR and iS-RESPDOR experiments.

  20. Application of solid state NMR for the study of surface bound species and fossil fuels

    NASA Astrophysics Data System (ADS)

    Althaus, Stacey

    Recent advances in solid state NMR have been utilized to study a variety of systems. These advancements have allowed for the acquisition of sequences previously only available for solution state detection. The protocol for the measurement of coals and other carbonaceous materials was updated to incorporate the recent advancements in fast magic angle spinning (MAS) and high magnetic fields. Argonne Premium Coals were used to test the sensitivity and resolution of the experiments preformed at high field and fast MAS. The higher field spectra were shown to be slightly less sensitive than the traditional lower field spectra, however, the new high field fast MAS spectra had better resolution. This increased resolution allowed for the separation of a variety of different functional groups, thereby allowing the composition of the coal to be determined. The use of 1 H detection allowed for 2D spectra of coals for the first time. These spectra could be filtered to examine either through-space or through-bond correlations. Indirect detection via 1 H was also pivotal in the detection of natural abundance 15 N spectra. Through-space and through-bond 2D spectra of natural abundance bulk species are shown with a sensitivity increase of 15 fold over traditional detection. This sensitivity enhancement allowed for the detection of natural abundance 15 N surface bound species in 2D, something that could not be acquired via traditional methods. The increased efficiency of the through-space magnetization transfer, Cross polarization, at fast MAS compared to the slower MAS rates is shown. The through-bond magnetization transfer via INEPT was examined and the effect of J-coupling is confirmed. Solid State NMR can be utilized to help improve catalytic interactions. Solid state NMR was used to examine the aldol condensation between p-nitrobenzaldehyde and acetone. The formation of a stable intermediate with p-nitrobenzaldehyde was found on the primary functionalized amine mesoporous nanoparticle when the reaction was done in hexane. When secondary amines were used to catalyze the reaction, the reactivity was increased and no intermediate was observed. When the solvent was changed from hexane to water, a dramatic increase in the reactivity of the primary amine was observed. The secondary amine was shown to have a large decrease in activity with the solvent change. The role of surface silanols in the catalysis with the mesoporous silica nanoparticles is also examined. Implementation of pulsed field gradient fields allowed for an examination of the diffusion of two different solvents , hexane and water, within the primary functionalized amine heterogeneous mesoporous catalyst system. The diffusion of the 2.7 nm pore sizes were compared to the diffusion in the 3.7 nm pores. In hexane the intra-pore diffusion in the smaller pores was slower than the larger pores, agreeing well with the low catalytic yield seen in the aldol reaction.

  1. In Situ Solid-State (13)C NMR Observation of Pore Mouth Catalysis in Etherification of β-Citronellene with Ethanol on Zeolite Beta.

    PubMed

    Radhakrishnan, Sambhu; Goossens, Pieter-Jan; Magusin, Pieter C M M; Sree, Sreeprasanth Pulinthanathu; Detavernier, Christophe; Breynaert, Eric; Martineau, Charlotte; Taulelle, Francis; Martens, Johan A

    2016-03-01

    The reaction mechanism of etherification of β-citronellene with ethanol in liquid phase over acid zeolite beta is revealed by in situ solid-state (13)C NMR spectroscopy. Comparison of (13)C Hahn-echo and (1)H-(13)C cross-polarization NMR characteristics is used to discriminate between molecules freely moving in liquid phase outside the zeolite and molecules adsorbed inside zeolite pores and in pore mouths. In the absence of ethanol, β-citronellene molecules enter zeolite pores and react to isomers. In the presence of ethanol, the concentration of β-citronellene inside zeolite pores is very low because of preferential adsorption of ethanol. The etherification reaction proceeds by adsorption of β-citronellene molecule from the external liquid phase in a pore opening where it reacts with ethanol from inside the pore. By competitive adsorption, ethanol prevents the undesired side reaction of β-citronellene isomerization inside zeolite pores. β-citronellene etherification on zeolite beta is suppressed by bulky base molecules (2,4,6-collidine and 2,6-ditertiarybutylpyridine) that do not enter the zeolite pores confirming the involvement of easily accessible acid sites in pore openings. The use of in situ solid-state NMR to probe the transition from intracrystalline catalysis to pore mouth catalysis depending on reaction conditions is demonstrated for the first time. The study further highlights the potential of this NMR approach for investigations of adsorption of multicomponent mixtures in general. PMID:26842944

  2. Solid-state NMR and IRMS characterization of smouldered peat from ombrotrophic cores

    NASA Astrophysics Data System (ADS)

    Zaccone, Claudio; Rein, Guillermo; Gioacchini, Paola; Knicker, Heike; Ciavatta, Claudio; Miano, Teodoro M.

    2013-04-01

    Smouldering fires are slow, low temperature, flameless and the most persistent form of combustion of organic matter (OM) in porous form. Although smouldering fires of peatlands represent a large perturbation of the atmospheric chemistry, to date, most studies on smouldering focused on ignition, carbon (C) losses or emissions, whereas the literature still lacks understanding of the OM evolution following these events. The potential to track OM changes able to serve as new proxies for the identification of past fire events along peat cores is extremely important, especially considering that bogs are often used as natural archives of paleoenvironmental changes. In the present work we show preliminary results about solid-state Nuclear Magnetic Resonance (NMR) and Isotope-Ratio Mass Spectroscopy (IRMS) characterization of peat OM along three Sphagnum peat columns (26 cm deep) having different initial moisture contents (MC): 50% MC, 100% MC, and 200% MC. The 15N spectrum of fresh peat (FP) used as control shows, as expected, only an amide signal, which is in agreement with the 13C NMR spectrum where mainly signals of carbohydrates and alkyl C can be observed. Further signals can be observed in the aromatic region, most probably due to lignin derivatives. Following the smouldering event, selected peat samples from both the 50% and 100% MC series show, as expected, signals supporting the occurrence of fire. In detail, the 15N-signals between -200 and -250 ppm are typical for pyrrole or indole type N. This is in accordance with the 13C NMR spectra showing considerable intensity in the aromatic region, most likely from char residues. Isotopic signatures (i.e., δ13C and δ15N) show a very interesting behaviour. In detail, δ13C seems to be slightly affected by smouldering, although the information about vegetational changes are preserved, whereas the δ15N shows a trend positively correlated with the relative N enrichment observed in smouldered peat samples, as also supported by 15N NMR. While further research is in progress to find reliable proxies allowing reconstruction of ancient smouldering events along peat profiles, our data provide an additional important insight towards assessing palaeoenvironmental conditions and highlighting that smouldering fires may have been overlooked as the cause of molecular and chemical variations observed in peat cores. The present research was financed by the Italian PRIN program 2009 (2009NBHPWR - Project title: "Chemical and biomolecular indicators for reconstructing environmental changes in natural archives")

  3. Biomolecular solid state NMR with magic-angle spinning at 25 K

    PubMed Central

    Thurber, Kent R.; Tycko, Robert

    2009-01-01

    A magic-angle spinning (MAS) probe has been constructed which allows the sample to be cooled with helium, while the MAS bearing and drive gases are nitrogen. The sample can be cooled to 25 K using roughly 3 liters/hour of liquid helium, while the 4 mm diameter rotor spins at 6.7 kHz with good stability (±5 Hz) for many hours. Proton decoupling fields up to at least 130 kHz can be applied. This helium-cooled MAS probe enables a variety of one-dimensional and two-dimensional NMR experiments on biomolecular solids and other materials at low temperatures, with signal-to-noise proportional to 1/T. We show examples of low-temperature 13C NMR data for two biomolecular samples, namely the peptide Aβ14–23 in the form of amyloid fibrils and the protein HP35 in frozen glycerol/water solution. Issues related to temperature calibration, spin-lattice relaxation at low temperatures, paramagnetic doping of frozen solutions, and 13C MAS NMR linewidths are discussed. PMID:18922715

  4. Structural Changes Associated with Transthyretin Misfolding and Amyloid Formation Revealed by Solution and Solid-State NMR.

    PubMed

    Lim, Kwang Hun; Dasari, Anvesh K R; Hung, Ivan; Gan, Zhehong; Kelly, Jeffery W; Wemmer, David E

    2016-04-01

    Elucidation of structural changes involved in protein misfolding and amyloid formation is crucial for unraveling the molecular basis of amyloid formation. Here we report structural analyses of the amyloidogenic intermediate and amyloid aggregates of transthyretin using solution and solid-state nuclear magnetic resonance (NMR) spectroscopy. Our solution NMR results show that one of the two main β-sheet structures (CBEF β-sheet) is maintained in the aggregation-competent intermediate, while the other DAGH β-sheet is more flexible on millisecond time scales. Magic-angle-spinning solid-state NMR revealed that AB loop regions interacting with strand A in the DAGH β-sheet undergo conformational changes, leading to the destabilized DAGH β-sheet. PMID:26998642

  5. High-Resolution NMR of Quadrupolar Nuclei in the Solid State

    SciTech Connect

    Gann, Sheryl Lee

    1995-11-30

    This dissertation describes recent developments in solid state nuclear magnetic resonance (NMR), for the most part involving the use of dynamic-angle spinning (DAS) NMR to study quadrupolar nuclei. Chapter 1 introduces some of the basic concepts and theory that will be referred to in later chapters, such as the density operator, product operators, rotations, coherence transfer pathways, phase cycling, and the various nuclear spin interactions, including the quadrupolar interaction. Chapter 2 describes the theory behind motional averaging experiments, including DAS, which is a technique where a sample is spun sequentially about two axis oriented at different angles with respect to the external magnetic field such that the chemical shift and quadrupolar anisotropy are averaged to zero. Work done on various rubidium-87 salts is presented as a demonstration of DAS. Chapter 3 explains how to remove sidebands from DAS and magic-angle spinning (MAS) experiments, which result from the time-dependence of the Hamiltonian under sample spinning conditions, using rotor-synchronized {pi}-pulses. Data from these experiments, known as DAH-180 and MAH-180, respectively, are presented for both rubidium and lead salts. In addition, the applicability of this technique to double rotation (DOR) experiments is discussed. Chapter 4 concerns the addition of cross-polarization to DAS (CPDAS). The theory behind spin locking and cross polarizing quadrupolar nuclei is explained and a method of avoiding the resulting problems by performing cross polarization at 0{sup o} (parallel) with respect to the magnetic field is presented. Experimental results are shown for a sodium-23 compound, sodium pyruvate, and for oxygen-17 labeled L-akmine. In Chapter 5, a method for broadening the Hartmann-Hahn matching condition under MAS, called variable effective field cross-polarization (VEFCI?), is presented, along with experimental work on adamantane and polycarbonate.

  6. Calcination products of gibbsite studied by X-ray diffraction, XPS and solid-state NMR

    SciTech Connect

    Malki, A.; Mekhalif, Z.; Detriche, S.; Fonder, G.; Boumaza, A.; Djelloul, A.

    2014-07-01

    The changes caused by heat treatment of gibbsite powder at 300–1473 K were studied using the X-ray diffraction (XRD), X-ray photoemission (XPS) spectra and {sup 27}Al magic angle spinning nuclear magnetic resonance spectroscopy ({sup 27}Al MAS NMR). XRD analysis indicates that the transformation sequence involves the formation of κ-Al{sub 2}O{sub 3} as an intermediate phase between χ- and α-Al{sub 2}O{sub 3}. The crystallite size of χ-Al{sub 2}O{sub 3} is as small as 10 nm. XPS analysis indicates that the ratio of aluminium atoms to oxygen atoms in χ-Al{sub 2}O{sub 3} and κ-Al{sub 2}O{sub 3} increases, whereas the expected ratio is observed in α-Al{sub 2}O{sub 3}. The percentage of AlO{sub 4} units in the transition aluminas follows the same behaviour as the ratio of Al/O. - Graphical abstract: The percentage of AlO{sub 4} units in transition aluminas follows the same behaviour as the ratio of Al/O. - Highlights: • Calcination products of gibbsite studied by XRD, XPS and solid-state NMR. • The crystallite size of χ-Al{sub 2}O{sub 3} is as small as 10 nm. • The Al/O atomic ratio determined by XPS is larger than 2/3 in χ-Al{sub 2}O{sub 3} and κ-Al{sub 2}O{sub 3}. • The percentage of AlO{sub 4} in the aluminas follows the same behaviour as the Al/O atomic ratio.

  7. Solid-state {sup 109}Ag NMR characterization of silver dispersed on oxide supports

    SciTech Connect

    Plischke, J.K.; Benesi, A.J.; Vannice, M.A.

    1992-04-30

    Magic angle spinning {sup 109}Ag solid-state NMR spectroscopy, transmission electron microscopy, X-ray diffraction, and gas adsorption measurements were used to characterize Ag particles supported on SiO{sub 2}, {eta}-Al{sub 2}O{sub 3}, and zeolite A. The samples were prepared either by a standard incipient wetness technique or by ion exchange followed by drying and reduction in H{sub 2}. {sup 109}Ag NMR spectroscopy was used to follow the state of the silver throughout the preparation process. {sup 109}Ag spectra were obtained for metallic Ag and for pure powder samples of AgNO{sub 3} and AgCl as well as for Ag{sup +} in solution; however, no resonances were observed for powder samples of Ag{sub 2}O and Ag{sub 2}O{sub 2}. Prior to drying, chemical shifts were found to be close to those of aqueous AgNO{sub 3} solutions ({approximately}O ppm), and after drying at 373 K, the chemical shift moved upfield to approach that of bulk AgNO{sub 3}. After reduction, single Knight-shifted metallic resonance at +5252{plus_minus} 10 ppm was observed for all samples which contained Ag particles larger than 50 nm in diameter. Low-loading, well-dispersed samples with smaller Ag particles yielded no observable {sup 109}Ag resonance. It is proposed that the absence of an observable {sup 109}Ag signal in the latter samples is due to a surface boundary effect on conduction electrons, common to other metal conductors, which broadens the signal. The adsorption of O{sub 2}, Cl{sub 2}, and HCl on the surface of the large Ag crystallites had no effect on the {sup 109}Ag spectrum. 33 refs., 10 figs., 2 tabs.

  8. High resolution 13C-detected solid-state NMR spectroscopy of a deuterated protein

    PubMed Central

    Tang, Ming; Comellas, Gemma; Mueller, Leonard J.

    2011-01-01

    High resolution 13C-detected solid-state NMR spectra of the deuterated beta-1 immunoglobulin binding domain of the protein G (GB1) have been collected to show that all 15N, 13C′, 13Cα and 13Cβ sites are resolved in 13C–13C and 15N–13C spectra, with significant improvement in T2 relaxation times and resolution at high magnetic field (750 MHz). The comparison of echo T2 values between deuterated and protonated GB1 at various spinning rates and under different decoupling schemes indicates that 13Cα T2′ times increase by almost a factor of two upon deuteration at all spinning rates and under moderate decoupling strength, and thus the deuteration enables application of scalar-based correlation experiments that are challenging from the standpoint of transverse relaxation, with moderate proton decoupling. Additionally, deuteration in large proteins is a useful strategy to selectively detect polar residues that are often important for protein function and protein–protein interactions. PMID:20803233

  9. Characterization of Stratum Corneum Molecular Dynamics by Natural-Abundance 13C Solid-State NMR

    PubMed Central

    Bouwstra, Joke A.; Sparr, Emma; Topgaard, Daniel

    2013-01-01

    Despite the enormous potential for pharmaceutical applications, there is still a lack of understanding of the molecular details that can contribute to increased permeability of the stratum corneum (SC). To investigate the influence of hydration and heating on the SC, we record the natural-abundance 13C signal of SC using polarization transfer solid-state NMR methods. Resonance lines from all major SC components are assigned. Comparison of the signal intensities obtained with the INEPT and CP pulse sequences gives information on the molecular dynamics of SC components. The majority of the lipids are rigid at 32°C, and those lipids co-exist with a small pool of mobile lipids. The ratio between mobile and rigid lipids increases with hydration. An abrupt change of keratin filament dynamics occurs at RH = 80–85%, from completely rigid to a structure with rigid backbone and mobile protruding terminals. Heating has a strong effect on the lipid mobility, but only a weak influence on the keratin filaments. The results provide novel molecular insight into how the SC constituents are affected by hydration and heating, and improve the understanding of enhanced SC permeability, which is associated with elevated temperatures and SC hydration. PMID:23626744

  10. 1H and 13C solid-state NMR of Gossypium barbadense (Pima) cotton

    NASA Astrophysics Data System (ADS)

    Taylor, R. E.; French, Alfred D.; Gamble, Gary R.; Himmelsbach, David S.; Stipanovic, Robert D.; Thibodeaux, Devron P.; Wakelyn, Phillip J.; Dybowski, C.

    2008-04-01

    The interaction of water with cellulose and its influence on the nuclear spin dynamics in Gossypium barbadense (Pima) cotton were investigated by 1H and 13C solid-state NMR techniques. 1H spin diffusion results from a Goldman-Shen experiment indicate that the water is multilayered. 1H MAS experiments provide evidence of a range of correlation times for the water, indicative of molecular motion ranging from restricted to relatively mobile. The 1H spin-lattice relaxation time varies with water content and is different for static and MAS conditions. By coupling the Goldman-Shen sequence with 13C CP/MAS, cross-polarization from the molecularly mobile water protons distributes magnetization throughout the cellulose (as opposed to enhancing 13C resonances from only the crystalline or the amorphous domains or from only the surface of the cellulose). However, spatial localization of the combined Goldman-Shen- 13C CP/MAS experiment using both short mixing and contact times yields a spectrum consistent with predominantly the I β polymorph of cellulose. Longer mixing times and the same, short contact time yield a spectrum that is indicative of an increased I α polymorph content in the crystallite interiors relative to the smaller values found with short mixing times.

  11. Solid-state NMR analysis of a boron-containing pharmaceutical hydrochloride salt.

    PubMed

    Vogt, Frederick G; Williams, Glenn R; Copley, Royston C B

    2013-10-01

    A novel crystalline form of the boron-containing antibacterial drug (S)-3-(aminomethyl)-7-(3-hydroxypropoxy)benzo[c] [1,2]oxaborol-1(3H)-ol hydrochloride is studied by solid-state nuclear magnetic resonance (SSNMR) and single-crystal X-ray diffraction techniques. After determination of the crystal structure by X-ray diffraction, SSNMR spectroscopy of this form is performed to obtain structural information using experimental approaches based on dipolar correlation, chemical shift analysis, and quadrupolar interaction analysis. 1H SSNMR experiments at 16.4 T using magic-angle spinning (MAS) and homonuclear dipolar decoupling, 2D SSNMR experiments based on (1)H–(13)C and (1)H–(11)B dipolar heteronuclear correlation, and density functional theory (DFT) calculations are combined in a novel approach to obtain a nearly complete assignment of the (1)H spectrum of this crystalline phase. (11)B and (35)Cl chemical shift and quadrupolar parameters are obtained using the analysis of MAS spectra and are found to be accurately reproduced using DFT calculations. NMR chemical shielding and electric field gradient parameters obtained using these methods are related to hydrogen-bonding trends in the crystal structure. The results illustrate the increasing capability of SSNMR techniques involving (1)H, (11)B, and (35)Cl SSNMR in the analysis of the crystal structure of a pharmaceutical compound containing covalently bonded boron. PMID:23918278

  12. Solid-state NMR studies of proteins immobilized on inorganic surfaces

    PubMed Central

    Shaw, Wendy J.

    2015-01-01

    Solid state NMR is the primary tool for studying the quantitative, site-specific structure, orientation, and dynamics of biomineralization proteins under biologically relevant conditions. Two calcium phosphate proteins, statherin (43 amino acids) and leucine rich amelogenin protein (LRAP; 59 amino acids), have been studied in depth and have different dynamic properties and 2D- and 3D-structural features. These differences make it difficult to extract design principles used in nature for building materials with properties such as high strength, unusual morphologies, or uncommon phases. Consequently, design principles needed for developing synthetic materials controlled by proteins are not clear. Many biomineralization proteins are much larger than statherin and LRAP, necessitating the study of larger biomineralization proteins. More recent studies of the significantly larger full-length amelogenin (180 residues) represent a significant step forward to ultimately investigate the full diversity of biomineralization proteins. Interactions of amino acids, a silaffin derived peptide, and the model LK peptide with silica are also being studied, along with qualitative studies of the organic matrices interacting with calcium carbonate. Dipolar recoupling techniques have formed the core of the quantitative studies, yet the need for isolated spin pairs makes this approach costly and time intensive. The use of multidimensional techniques to study biomineralization proteins is becoming more common, methodology which, despite its challenges with these difficult-to-study proteins, will continue to drive future advancements in this area. PMID:25466354

  13. Exploring Dynamics and Cage-Guest Interactions in Clathrate Hydrates Using Solid-State NMR.

    PubMed

    Sengupta, Suvrajit; Guo, Jin; Janda, Kenneth C; Martin, Rachel W

    2015-12-17

    Interactions between guest molecules and the water cages in clathrates are dominated by isotropic van der Waals forces at low temperatures because the cage structures satisfy the hydrogen bonding propensity of the water. However, above 200 K the water molecules become more labile and may interact strongly with the guests through hydrogen bonding. In this work we compare the dynamics of tetrahydrofuran (THF) and cyclopentane (CP) guests in the hydrate cages above 200 K. Since THF can form hydrogen bonds while CP cannot, the dynamics provide insight into host-guest hydrogen bonding. We use magic angle spinning (MAS) solid-state NMR to measure proton spin-lattice relaxation times (T1) of the guests as a function of temperature and find that the activation barrier to the motion of THF molecules is 4.7 kcal/mol (19.7 kJ/mol) at temperatures above 200 K. This is almost 5 times higher than the barrier at lower temperatures. In contrast, the barrier to guest motion in CP hydrate is found to be about 0.67 kcal/mol (2.8 kJ/mol), which agrees well with data at lower temperatures. These results demonstrate that hydrogen bonding interactions between the THF guest and the clathrate cage are significant above 200 K due to the host lattice mobility. PMID:26583257

  14. Self-Assembly of X-Shaped Bolapolyphiles in Lipid Membranes: Solid-State NMR Investigations.

    PubMed

    Achilles, Anja; Bärenwald, Ruth; Lechner, Bob-Dan; Werner, Stefan; Ebert, Helgard; Tschierske, Carsten; Blume, Alfred; Bacia, Kirsten; Saalwächter, Kay

    2016-01-26

    A novel class of rigid-rod bolapolyphilic molecules with three philicities (rigid aromatic core, mobile aliphatic side chains, polar end groups) has recently been demonstrated to incorporate into and span lipid membranes, and to exhibit a rich variety of self-organization modes, including macroscopically ordered snowflake structures with 6-fold symmetry. In order to support a structural model and to better understand the self-organization on a molecular scale, we here report on proton and carbon-13 high-resolution magic-angle spinning solid-state NMR investigations of two different bolapolyphiles (BPs) in model membranes of two different phospholipids (DPPC, DOPC). We elucidate the changes in molecular dynamics associated with three new phase transitions detected by calorimetry in composite membranes of different composition, namely, a change in π-π-packing, the melting of lipid tails associated with the superstructure, and the dissolution and onset of free rotation of the BPs. We derive dynamic order parameters associated with different H-H and C-H bond directions of the BPs, demonstrating that the aromatic cores are well packed below the final phase transition, showing only 180° flips of the phenyl ring, and that they perform free rotations with additional oscillations of the long axis when dissolved in the fluid membrane. Our data suggests that BPs not only form ordered superstructures, but also rather homogeneously dispersed π-packed filaments within the lipid gel phase, thus reducing the corrugation of large vesicles. PMID:26735449

  15. Molecular dynamics of neutral polymer bonding agent (NPBA) as revealed by solid-state NMR spectroscopy.

    PubMed

    Hu, Wei; Su, Yongchao; Zhou, Lei; Pang, Aimin; Cai, Rulin; Ma, Xingang; Li, Shenhui

    2013-01-01

    Neutral polymer bonding agent (NPBA) is one of the most promising polymeric materials, widely used in nitrate ester plasticized polyether (NEPE) propellant as bonding agent. The structure and dynamics of NPBA under different conditions of temperatures and sample processing are comprehensively investigated by solid state NMR (SSNMR). The results indicate that both the main chain and side chain of NPBA are quite rigid below its glass transition temperature (Tg). In contrast, above the Tg, the main chain remains relatively immobilized, while the side chains become highly flexible, which presumably weakens the interaction between bonding agent and the binder or oxidant fillers and in turn destabilizes the high modulus layer formed around the oxidant fillers. In addition, no obvious variation is found for the microstructure of NPBA upon aging treatment or soaking with acetone. These experimental results provide useful insights for understanding the structural properties of NPBA and its interaction with other constituents of solid composite propellants under different processing and working conditions. PMID:24451254

  16. Structure and backbone dynamics of a microcrystalline metalloprotein by solid-state NMR

    PubMed Central

    Knight, Michael J.; Pell, Andrew J.; Bertini, Ivano; Felli, Isabella C.; Gonnelli, Leonardo; Pierattelli, Roberta; Herrmann, Torsten; Emsley, Lyndon; Pintacuda, Guido

    2012-01-01

    We introduce a new approach to improve structural and dynamical determination of large metalloproteins using solid-state nuclear magnetic resonance (NMR) with 1H detection under ultrafast magic angle spinning (MAS). The approach is based on the rapid and sensitive acquisition of an extensive set of 15N and 13C nuclear relaxation rates. The system on which we demonstrate these methods is the enzyme Cu, Zn superoxide dismutase (SOD), which coordinates a Cu ion available either in Cu+ (diamagnetic) or Cu2+ (paramagnetic) form. Paramagnetic relaxation enhancements are obtained from the difference in rates measured in the two forms and are employed as structural constraints for the determination of the protein structure. When added to 1H-1H distance restraints, they are shown to yield a twofold improvement of the precision of the structure. Site-specific order parameters and timescales of motion are obtained by a Gaussian axial fluctuation (GAF) analysis of the relaxation rates of the diamagnetic molecule, and interpreted in relation to backbone structure and metal binding. Timescales for motion are found to be in the range of the overall correlation time in solution, where internal motions characterized here would not be observable. PMID:22723345

  17. 1H and 13C NMR study of cyclopentadienyl metal carbonyls in the solid state

    NASA Astrophysics Data System (ADS)

    Aime, S.; Cordero, L.; Gobetto, R.; Szalontai, G.

    1993-08-01

    In this paper we deal with some structural and dynamic properties of Cp 2W 2(CO) 6 ( I) and Cp 2Ru 2(CO) 4 ( II) as shown by solid state 13C and 1H NMR experiments. The IR and 13C CPMAS spectra of a polycrystalline sample of I show that this compound possesses the anti rotameric structure found in a previously reported X-ray diffraction study. The analysis of the spinning side-band manifold in the 13C CPMAS spectrum of I allows us to assess a different semi-bridging character between two CO-groups not seen from the X-ray results. The spectral features of compound II are fully consistent with the X-ray and solution structures previously reported. In both compounds the cyclopentadienyl ligands are involved in fast reorientation motions which modulate the magnetic interactions responsible for the relaxation of 13C resonances. The activation energies ( Ea) associated with this reorientation process of the Cp ring along their C 5 coordination axis have been determined to be 15.5 and 10.2 kJ mol -1 for I and II respectively on the basis of 1H T1 measurements at different temperatures. Furthermore, we show that an empirical relationship relates Ea values and Tmin (the temperature at which proton relaxation is more efficient) in a related series of cyclopentadienyl compounds.

  18. Toward Relatively General and Accurate Quantum Chemical Predictions of Solid-State (17)O NMR Chemical Shifts in Various Biologically Relevant Oxygen-Containing Compounds.

    PubMed

    Rorick, Amber; Michael, Matthew A; Yang, Liu; Zhang, Yong

    2015-09-01

    Oxygen is an important element in most biologically significant molecules, and experimental solid-state (17)O NMR studies have provided numerous useful structural probes to study these systems. However, computational predictions of solid-state (17)O NMR chemical shift tensor properties are still challenging in many cases, and in particular, each of the prior computational works is basically limited to one type of oxygen-containing system. This work provides the first systematic study of the effects of geometry refinement, method, and basis sets for metal and nonmetal elements in both geometry optimization and NMR property calculations of some biologically relevant oxygen-containing compounds with a good variety of XO bonding groups (X = H, C, N, P, and metal). The experimental range studied is of 1455 ppm, a major part of the reported (17)O NMR chemical shifts in organic and organometallic compounds. A number of computational factors toward relatively general and accurate predictions of (17)O NMR chemical shifts were studied to provide helpful and detailed suggestions for future work. For the studied kinds of oxygen-containing compounds, the best computational approach results in a theory-versus-experiment correlation coefficient (R(2)) value of 0.9880 and a mean absolute deviation of 13 ppm (1.9% of the experimental range) for isotropic NMR shifts and an R(2) value of 0.9926 for all shift-tensor properties. These results shall facilitate future computational studies of (17)O NMR chemical shifts in many biologically relevant systems, and the high accuracy may also help the refinement and determination of active-site structures of some oxygen-containing substrate-bound proteins. PMID:26274812

  19. Elucidating Metabolic Pathways for Amino Acid Incorporation Into Dragline Spider Silk using 13C Enrichment and Solid State NMR

    PubMed Central

    Creager, Melinda S.; Izdebski, Thomas; Brooks, Amanda E.; Lewis, Randolph V.

    2013-01-01

    Spider silk has been evolutionarily optimized for contextual mechanical performance over the last 400 million years. Despite precisely balanced mechanical properties, which have yet to be reproduced, the underlying molecular architecture of major ampullate spider silk can be simplified being viewed as a versatile block copolymer. Four primary amino acid motifs: polyalanine, (GA)n, GPGXX, and GGX (X = G,A,S,Q,L,Y) will be considered in this study. Although synthetic mimetics of many of these amino acid motifs have been produced in several biological systems, the source of spider silk’s mechanical integrity remains elusive. Mechanical robustness may be a product not only of the amino acid structure but also of the tertiary structure of the silk. Historically, solid state Nuclear Magnetic Resonance (ssNMR) has been used to reveal the crystalline structure of the polyalanine motif; however, limitations in amino acid labeling techniques have obscured the structures of the GGX and GPGXX motifs thought to be responsible for the structural mobility of spider silk. We describe the use of metabolic pathways to label tyrosine for the first time as well as to improve the labeling efficiency of proline. These improved labeling techniques will allow the previously unknown tertiary structures of major ampullate silk to be probed. PMID:21334448

  20. Probing the Small-Molecule Inhibition of an Anticancer Therapeutic Protein-Protein Interaction Using a Solid-State Nanopore.

    PubMed

    Kwak, Dong-Kyu; Chae, Hongsik; Lee, Mi-Kyung; Ha, Ji-Hyang; Goyal, Gaurav; Kim, Min Jun; Kim, Ki-Bum; Chi, Seung-Wook

    2016-05-01

    Nanopore sensing is an emerging technology for the single-molecule-based detection of various biomolecules. In this study, we probed the anticancer therapeutic p53 transactivation domain (p53TAD)/MDM2 interaction and its inhibition with a small-molecule MDM2 antagonist, Nutlin-3, using low-noise solid-state nanopores. Although the translocation of positively charged MDM2 through a nanopore was detected at the applied negative voltage, this MDM2 translocation was almost completely blocked upon formation of the MDM2/GST-p53TAD complex owing to charge conversion. In combination with NMR data, the nanopore measurements showed that the addition of Nutlin-3 rescued MDM2 translocation, indicating that Nutlin-3 disrupted the MDM2/GST-p53TAD complex, thereby releasing MDM2. Taken together, our results reveal that solid-state nanopores can be a valuable platform for the ultrasensitive, picomole-scale screening of small-molecule drugs against protein-protein interaction (PPI) targets. PMID:27038437

  1. (13) C-TmDOTA as versatile thermometer compound for solid-state NMR of hydrated lipid bilayer membranes.

    PubMed

    Umegawa, Yuichi; Tanaka, Yuya; Nobuaki, Matsumori; Murata, Michio

    2016-03-01

    Recent advances in solid-state nuclear magnetic resonance (NMR) techniques, such as magic angle spinning and high-power decoupling, have dramatically increased the sensitivity and resolution of NMR. However, these NMR techniques generate extra heat, causing a temperature difference between the sample in the rotor and the variable temperature gas. This extra heating is a particularly crucial problem for hydrated lipid membrane samples. Thus, to develop an NMR thermometer that is suitable for hydrated lipid samples, thulium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (TmDOTA) was synthesized and labeled with (13) C (i.e., (13) C-TmDOTA) to increase the NMR sensitivity. The complex was mixed with a hydrated lipid membrane, and the system was subjected to solid-state NMR and differential scanning calorimetric analyses. The physical properties of the lipid bilayer and the quality of the NMR spectra of the membrane were negligibly affected by the presence of (13) C-TmDOTA, and the (13) C chemical shift of the complex exhibited a large-temperature dependence. The results demonstrated that (13) C-TmDOTA could be successfully used as a thermometer to accurately monitor temperature changes induced by (1) H decoupling pulses and/or by magic angle spinning and the temperature distribution of the sample inside the rotor. Thus, (13) C-TmDOTA was shown to be a versatile thermometer for hydrated lipid assemblies. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26460094

  2. Thermal decomposition of t-butylamine borane studied by in situ solid state NMR

    SciTech Connect

    Feigerle, J.; Smyrl, N. R.; Morrell, J. S.; Stowe, A. C.

    2010-03-18

    Development of suitable materials to store hydrogen for automotive use has received pointed attention over the past decade. Significant progress has been made with the discovery of novel chemical hydrides, complex metal hydrides, and adsorption substrates which continue to optimize both thermodynamics and kinetics of hydrogen sorption. Chemical hydrides typically offer the largest theoretical gravimetric capacities. Autrey et al. have recently shown that mechanical milling of alkali metal hydrides with ammonia borane can further lower the decomposition temperature. In all cases, however, many challenges remain in order to meet the current US DOE performance targets. Amine boranes are being considered for hydrogen storage materials since they contain significant quantities of hydrogen which potentially can be released at low temperatures (80-150 C) via chemical reactions. Ammonia borane, NH{sub 3}BH{sub 3}, is one of the most promising in this class as it decomposes to release greater than two moles of pure hydrogen gas (14 wt%) below 160 C. Although isoelectronic to ethane, NH{sub 3}BH{sub 3} is a solid at room temperature due to the di-hydrogen bonding network formed between the amine protons and boron hydrides in the solid state lattice. Further, it has been shown that the hydrogen release mechanism involves transformation and isomerization to an ionic dimer where a hydride migrates from one boron to the adjacent boron in the dimer. The greatest challenge to the use of ammonia borane as a hydrogen fuel is the regeneration path from spent fuel to ammonia borane again. The proposed chemical synthesis involves complicated organometallic reactions to form boron hydrogen bonds from the thermodynamically stable polyimidoborane products (BNH){sub n}. Recent theoretical calculations suggested that incorporation of carbon atoms into the (BNH)n product would be less thermodynamically stable. These (CBNH)n compounds are potentially less energy intensive making regeneration of the amine borane fuel more feasible [22]. In the present study, tert-butylamine borane is investigated by heteronuclear in situ solid state NMR to understand hydrogen release from a hydrocarbon containing amine borane. tbutylamine borane has similar physical properties to amine borane with a melting point of 96 C. A single proton has been replaced with a t-butylamine group resulting in a weakening of the dihydrogen bonding framework. t-butylamine borane has a theoretical gravimetric hydrogen density of 15.1%; however, isobutane can also be evolved rather than hydrogen. If decomposition yields one mole isobutane and two moles hydrogen, 4.5 wt% H2 gas will be evolved. More importantly for the present work, the resulting spent fuel should be comprised of both (BNH)n and (CBNH)n polyimidoboranes.

  3. New insights into the bonding arrangements of L- and D-glutamates from solid state 17O NMR

    NASA Astrophysics Data System (ADS)

    Lemaitre, V.; Pike, K. J.; Watts, A.; Anupold, T.; Samoson, A.; Smith, M. E.; Dupree, R.

    2003-03-01

    Magic angle spinning (MAS) from L- and D-glutamic acid-HCl at 14.1 T produces highly structured and very similar NMR spectra. Lines from all 4 oxygen sites are readily distinguished and assigned. These 17O NMR spectra are very different from the previously reported 17O spectrum of the D, L-form presumably because that was a racemic crystal. 17O NMR from L-monosodium glutamate-HCl is very different again requiring the application of double angle rotation and 3 quantum MAS NMR to provide resolution of 5 different sites. Hence high resolution 17O solid state NMR techniques offer possible new insight into biochemical bonding processes.

  4. Solid-state NMR enhanced by dynamic nuclear polarization as a novel tool for ribosome structural biology.

    PubMed

    Gelis, Ioannis; Vitzthum, Veronika; Dhimole, Neha; Caporini, Marc A; Schedlbauer, Andreas; Carnevale, Diego; Connell, Sean R; Fucini, Paola; Bodenhausen, Geoffrey

    2013-06-01

    The impact of Nuclear Magnetic Resonance (NMR) on studies of large macromolecular complexes hinges on improvements in sensitivity and resolution. Dynamic nuclear polarization (DNP) in the solid state can offer improved sensitivity, provided sample preparation is optimized to preserve spectral resolution. For a few nanomoles of intact ribosomes and an 800 kDa ribosomal complex we demonstrate that the combination of DNP and magic-angle spinning NMR (MAS-NMR) allows one to overcome current sensitivity limitations so that homo- and heteronuclear (13)C and (15)N NMR correlation spectra can be recorded. Ribosome particles, directly pelleted and frozen into an NMR rotor, yield DNP signal enhancements on the order of ~25-fold and spectra that exhibit narrow linewidths, suitable for obtaining site-specific information. We anticipate that the same approach is applicable to other high molecular weight complexes. PMID:23689811

  5. Solid-state NMR/NQR and first-principles study of two niobium halide cluster compounds.

    PubMed

    Perić, Berislav; Gautier, Régis; Pickard, Chris J; Bosiočić, Marko; Grbić, Mihael S; Požek, Miroslav

    2014-01-01

    Two hexanuclear niobium halide cluster compounds with a [Nb6X12](2+) (X=Cl, Br) diamagnetic cluster core, have been studied by a combination of experimental solid-state NMR/NQR techniques and PAW/GIPAW calculations. For niobium sites the NMR parameters were determined by using variable Bo field static broadband NMR measurements and additional NQR measurements. It was found that they possess large positive chemical shifts, contrary to majority of niobium compounds studied so far by solid-state NMR, but in accordance with chemical shifts of (95)Mo nuclei in structurally related compounds containing [Mo6Br8](4+) cluster cores. Experimentally determined δiso((93)Nb) values are in the range from 2,400 to 3,000 ppm. A detailed analysis of geometrical relations between computed electric field gradient (EFG) and chemical shift (CS) tensors with respect to structural features of cluster units was carried out. These tensors on niobium sites are almost axially symmetric with parallel orientation of the largest EFG and the smallest CS principal axes (Vzz and δ33) coinciding with the molecular four-fold axis of the [Nb6X12](2+) unit. Bridging halogen sites are characterized by large asymmetry of EFG and CS tensors, the largest EFG principal axis (Vzz) is perpendicular to the X-Nb bonds, while intermediate EFG principal axis (Vyy) and the largest CS principal axis (δ11) are oriented in the radial direction with respect to the center of the cluster unit. For more symmetrical bromide compound the PAW predictions for EFG parameters are in better correspondence with the NMR/NQR measurements than in the less symmetrical chlorine compound. Theoretically predicted NMR parameters of bridging halogen sites were checked by (79/81)Br NQR and (35)Cl solid-state NMR measurements. PMID:24581866

  6. Rheological and solid-state NMR assessments of copovidone/clotrimazole model solid dispersions.

    PubMed

    Yang, Fengyuan; Su, Yongchao; Zhu, Lei; Brown, Chad D; Rosen, Lawrence A; Rosenberg, Kenneth J

    2016-03-16

    This study aims to assess several model solid dispersions by using dynamic oscillatory rheology, solid-state NMR and other solid phase characterization techniques, and correlate their viscoelastic responses with processing methods and microstructures. A model active pharmaceutical ingredient (API), clotrimazole, was compounded with copovidone to form solid dispersions via various techniques with different mixing capabilities. Physicochemical characterizations of the resulting solid dispersions demonstrated that simple physical mixing led to a poorly mixed blend manifested by existence of large API crystalline content and heterogeneous distribution. Cryogenic milling significantly improved mixing of two components as a result of reduced particle size and increased contact surface area, but produced limited amorphous content. In contrast, hot melt extrusion (HME) processing resulted in a homogenous amorphous solid dispersion because of its inherent mixing efficiency. Storage modulus and viscosities versus frequency of different solid dispersions indicated that the incorporation of API into the polymer matrix resulted in a plasticizing effect which reduced the viscosity. The crystalline/aggregated forms of API also exhibited more elastic response than its amorphous/dispersed counterpart. Temperature ramps of the physical mixture with high API concentration captured a critical temperature, at which a bump was observed in damping factor. This bump was attributed to the dissolution of crystalline API into the polymer. In addition, heating-cooling cycles of various solid dispersions suggested that cryomilling and HME processing could form a homogeneous solid dispersion at low API content, whereas high drug concentration led to a relatively unstable dispersion due to supersaturation of API in the polymer. PMID:26780122

  7. 13C solid-state NMR analysis of heterogeneous structure of beeswax in native state

    NASA Astrophysics Data System (ADS)

    Kameda, Tsunenori

    2005-12-01

    I investigated the molecular structure of natural wax from Japanese bees (Apis cerana japonica) in its native state (neither purified nor recrystallized) by 13C and 1H solid-state NMR. Two strong 13C peaks at 32.9 and 34.0 ppm were attributed to signals from internal-chain methylene carbons [int-(CH2)] in two types of crystal form. The peak at 32.9 ppm was assigned to an orthorhombic crystal form, and that at 34.0 ppm was assigned to a triclinic or monoclinic form. In both crystalline regions, bi-exponential decay of 13C spin-lattice relaxation [T1(C)] for the crystalline peaks due to chain diffusion was observed. 1H spin-lattice relaxation [T1(H)] values for protons of the CH3 group and for int-(CH2) in the crystalline and amorphous regions were identical; this was interpreted as being due to averaging of the T1(H) relaxation rates via spin diffusion. In contrast, although the T_{{1}_{\\rho}}(H) decay curves for protons of the CH3 group and for int-(CH2) in the amorphous and orthorhombic forms were almost identical, those of the triclinic or monoclinic forms were different. This unhomogeneous character of T_{{1}_{\\rho}}(H) was interpreted as resulting from differences in the molecular composition of each crystal form. Moreover, two components with long and short 1H spin-spin relaxation [T2(H)] values, arising from the mobile and rigid phases, respectively, were observed at above about -30 °C.

  8. Characterization of the glass-to-vitroceramic transition in yttrium aluminum borate laser glasses using solid state NMR.

    PubMed

    Deters, Heinz; Eckert, Hellmut

    2012-02-01

    The crystallization of laser glasses in the system (Y(2)O(3))(0.2){((Al(2)O(3))(x))(B(2)O(3))(0.8-x)} (0.15 ≤ x ≤ 0.40) doped with 0.5 mol% of ytterbium oxide has been investigated by x-ray powder diffraction, and various solid state NMR techniques. The crystallization process has been analyzed in a quantitative fashion by high-resolution solid state (11)B, (27)Al, and (89)Y NMR spectroscopy as well as (11)B{(27)Al} and (27)Al{(11)B} rotational echo double resonance (REDOR) experiments. The homogeneous glasses undergo major phase segregation processes resulting in crystalline Al(5)BO(9) (historically denoted as Al(18)B(4)O(33)), YBO(3), crystalline YAl(3)(BO(3))(4), residual glassy B(2)O(3), and an additional yet not identified crystalline phase ("X-phase"). PMID:22244244

  9. Solid state NMR spectroscopy investigation of the molecular structure of epoxy based materials cured in different conditions

    NASA Astrophysics Data System (ADS)

    Alessi, S.; Spinella, A.; Caponetti, E.; Sabatino, Maria Antonietta; Spadaro, G.

    2012-07-01

    In this work two epoxy resin model systems, whose monomers are typically used in structural composites, were thermally cured in different cure conditions in order to obtain different cross-linking densities. Their molecular structures were investigated through solid state NMR spectroscopy in order to correlate them to the cure process conditions used and the results were discussed in the light of the dynamical mechanical thermal analysis (DMTA) performed.

  10. Quantitative determination of lead in mixtures of lead(II) halides using solid-state 207Pb NMR spectroscopy.

    PubMed

    Glatfelter, Alicia; Stephenson, Nicole; Bai, Shi; Dybowski, Cecil; Perry, Dale L

    2006-11-01

    We demonstrate a multi-spectrum technique for facile, quantitative determination of lead in solid materials using solid-state (207)Pb NMR that avoids the major problem of uniform excitation across a wide spectral range; the method can be employed without chemical separation or other chemical manipulations and without any prior sample preparation, resulting in a non-destructive analysis, and producing results that are in agreement with gravimetric analyses of mixed samples of the lead halides. PMID:17066187

  11. A natural abundance 33S solid-state NMR study of layered transition metal disulfides at ultrahigh magnetic field.

    PubMed

    Sutrisno, Andre; Terskikh, Victor V; Huang, Yining

    2009-01-01

    Using a series of layered transition metal disulfides we demonstrate that the wide-line natural abundance solid-state NMR spectra of 33S in a less symmetric environment can readily be obtained at ultrahigh magnetic field of 21.1 T and that surprisingly these closely related materials display a wide range of 33S quadrupole coupling constant and chemical shift anisotropy values. PMID:19099063

  12. Sodium ion effect on silk fibroin conformation characterized by solid-state NMR and generalized 2D NMR NMR correlation

    NASA Astrophysics Data System (ADS)

    Ruan, Qing-Xia; Zhou, Ping

    2008-07-01

    In the present work, we investigated Na + ion effect on the silk fibroin (SF) conformation. Samples are Na +-involved regenerated silk fibroin films. 13C CP-MAS NMR demonstrates that as added [Na +] increases, partial silk fibroin conformation transit from helix-form to β-form at certain Na + ion concentration which is much higher than that in Bombyx mori silkworm gland. The generalized two-dimensional NMR-NMR correlation analysis reveals that silk fibroin undergoes several intermediate states during its conformation transition process as [Na +] increase. The appearance order of the intermediates is followed as: helix and/or random coil → helix-like → β-sheet-like → β-sheet, which is the same as that produced by pH decrease from 6.8 to 4.8 in the resultant regenerated silk fibroin films. The binding sites of Na + to silk fibroin might involve the carbonyl oxygen atom of certain amino acids sequence which could promote the formation of β-sheet conformation. Since the Na +sbnd O bond is weak, the ability of Na + inducing the secondary structure transition is weaker than those of Ca 2+, Cu 2+ and even K +. It is maybe a reason why the sodium content is much lower than potassium in the silkworm gland.

  13. Morphology and molecular dynamics of hard ?-keratin under pressure by 1H and 13C solid-state NMR

    NASA Astrophysics Data System (ADS)

    Demco, Dan E.; Utiu, Lavinia; Tillmann, Walter; Blmich, Bernhard; Popescu, Crisan

    2011-06-01

    The effect of uniaxial pressure on phase composition, aminoacid side-chain and backbone dynamics, as well as rigid domain sizes of hard ?-keratin from human fingernail clippings was investigated by 1H solid-state and 13C cross-polarization MAS NMR spectroscopy. Proton spin-diffusion NMR experiments revealed that the rigid-domain sizes increased upon compression. The 13C carbonyl resonance components were shown to be very sensitive to the transition of ?-helices to ?-sheets induced by uniaxial compression. Carbon-13 longitudinal relaxation in the rotating frame showed dynamic heterogeneity of aminoacid residues especially during the ?-helix to ?-sheet transition.

  14. Solid-state NMR sequential assignments of the N-terminal domain of HpDnaB helicase.

    PubMed

    Wiegand, Thomas; Gardiennet, Carole; Ravotti, Francesco; Bazin, Alexandre; Kunert, Britta; Lacabanne, Denis; Cadalbert, Riccardo; Güntert, Peter; Terradot, Laurent; Böckmann, Anja; Meier, Beat H

    2016-04-01

    We present solid-state NMR assignments of the N-terminal domain of the DnaB helicase from Helicobacter pylori (153 residues) in its microcrystalline form. We use a sequential resonance assignment strategy based on three-dimensional NMR experiments. The resonance assignments obtained are compared with automated resonance assignments computed with the ssFLYA algorithm. An analysis of the (13)C secondary chemical shifts determines the position of the secondary structure elements in this α-helical protein. PMID:26280528

  15. Chemical structures of pyridine extracts and residues of coals as indicated by solid state 13C NMR.

    NASA Astrophysics Data System (ADS)

    Erbatur, G.; Erbatur, O.; Davis, M. F.; Maciel, G. E.

    1984-03-01

    The pyridine extracts and the residues of four Turkish coals were investigated in the solid state by CP/MAS C-13 NMR. No systematic correlation was observed between the ranks of the coals and the aromaticities of the pyridine extracts. There were pronounced structural differences between the pyridine extracts and the corresponding parent fuels and these differences were most prominent in the lower rank coals. In general, the C-13 NMR spectra of the residues were quite similar to those of the corresponding parent fuels.

  16. Lipid bilayer preparations of membrane proteins for oriented and magic-angle spinning solid-state NMR samples

    PubMed Central

    Das, Nabanita; Murray, Dylan T; Cross, Timothy A

    2014-01-01

    Solid-state NMR spectroscopy has been used successfully for characterizing the structure and dynamics of membrane proteins as well as their interactions with other proteins in lipid bilayers. such an environment is often necessary for achieving native-like structures. sample preparation is the key to this success. Here we present a detailed description of a robust protocol that results in high-quality membrane protein samples for both magic-angle spinning and oriented-sample solid-state NMR. the procedure is demonstrated using two proteins: CrgA (two transmembrane helices) and rv1861 (three transmembrane helices), both from Mycobacterium tuberculosis. the success of this procedure relies on two points. First, for samples for both types of NMR experiment, the reconstitution of the protein from a detergent environment to an environment in which it is incorporated into liposomes results in ‘complete’ removal of detergent. second, for the oriented samples, proper dehydration followed by rehydration of the proteoliposomes is essential. By using this protocol, proteoliposome samples for magic-angle spinning NMR and uniformly aligned samples (orientational mosaicity of <1°) for oriented-sample NMR can be obtained within 10 d. PMID:24157546

  17. Molecular characterization and quantification using state of the art solid-state adiabatic TOBSY NMR in burn trauma

    PubMed Central

    RIGHI, VALERIA; ANDRONESI, OVIDIU; MINTZOPOULOS, DIONYSSIOS; TZIKA, A. ARIA

    2009-01-01

    We describe a novel solid-state nuclear magnetic resonance (NMR) method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS), relative conventional liquid-state NMR approaches, when applied to intact biopsies of skeletal muscle specimens collected from burn trauma patients. This novel method, termed optimized adiabatic TOtal through Bond correlation SpectroscopY (TOBSY) solid-state NMR pulse sequence for two-dimensional (2D)1H-1H homonuclear scalar-coupling longitudinal isotropic mixing, was demonstrated to provide a 40–60% improvement in signal-to-noise ratio (SNR) relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). Using 1-and 2-dimensional HRMAS NMR experiments, we identified several metabolites in burned tissues. Quantification of metabolites in burned tissues showed increased levels of lipid compounds, intracellular metabolites (e.g., taurine and phosphocreatine) and substantially decreased water-soluble metabolites (e.g., glutathione, carnosine, glucose, glutamine/glutamate and alanine). These findings demonstrate that HRMAS NMR Spectroscopy using TOBSY is a feasible technique that reveals new insights into the pathophysiology of burn trauma. Moreover, this method has applications that facilitate the development of novel therapeutic strategies. PMID:19885614

  18. The contribution of solid-state NMR spectroscopy to understanding biomineralization: Atomic and molecular structure of bone

    NASA Astrophysics Data System (ADS)

    Duer, Melinda J.

    2015-04-01

    Solid-state NMR spectroscopy has had a major impact on our understanding of the structure of mineralized tissues, in particular bone. Bone exemplifies the organic-inorganic composite structure inherent in mineralized tissues. The organic component of the extracellular matrix in bone is primarily composed of ordered fibrils of collagen triple-helical molecules, in which the inorganic component, calcium phosphate particles, composed of stacks of mineral platelets, are arranged around the fibrils. This perspective argues that key factors in our current structural model of bone mineral have come about through NMR spectroscopy and have yielded the primary information on how the mineral particles interface and bind with the underlying organic matrix. The structure of collagen within the organic matrix of bone or any other structural tissue has yet to be determined, but here too, this perspective shows there has been real progress made through application of solid-state NMR spectroscopy in conjunction with other techniques. In particular, NMR spectroscopy has highlighted the fact that even within these structural proteins, there is considerable dynamics, which suggests that one should be cautious when using inherently static structural models, such as those arising from X-ray diffraction analyses, to gain insight into molecular roles. It is clear that the NMR approach is still in its infancy in this area, and that we can expect many more developments in the future, particularly in understanding the molecular mechanisms of bone diseases and ageing.

  19. Multiple acquisition of magic angle spinning solid-state NMR experiments using one receiver: Application to microcrystalline and membrane protein preparations

    NASA Astrophysics Data System (ADS)

    Gopinath, T.; Veglia, Gianluigi

    2015-04-01

    Solid-state NMR spectroscopy of proteins is a notoriously low-throughput technique. Relatively low-sensitivity and poor resolution of protein samples require long acquisition times for multidimensional NMR experiments. To speed up data acquisition, we developed a family of experiments called Polarization Optimized Experiments (POE), in which we utilized the orphan spin operators that are discarded in classical multidimensional NMR experiments, recovering them to allow simultaneous acquisition of multiple 2D and 3D experiments, all while using conventional probes with spectrometers equipped with one receiver. POE allow the concatenation of multiple 2D or 3D pulse sequences into a single experiment, thus potentially combining all of the aforementioned advances, boosting the capability of ssNMR spectrometers at least two-fold without the addition of any hardware. In this perspective, we describe the first generation of POE, such as dual acquisition MAS (or DUMAS) methods, and then illustrate the evolution of these experiments into MEIOSIS, a method that enables the simultaneous acquisition of multiple 2D and 3D spectra. Using these new pulse schemes for the solid-state NMR investigation of biopolymers makes it possible to obtain sequential resonance assignments, as well as distance restraints, in about half the experimental time. While designed for acquisition of heteronuclei, these new experiments can be easily implemented for proton detection and coupled with other recent advancements, such as dynamic nuclear polarization (DNP), to improve signal to noise. Finally, we illustrate the application of these methods to microcrystalline protein preparations as well as single and multi-span membrane proteins reconstituted in lipid membranes.

  20. Multiple acquisition of magic angle spinning solid-state NMR experiments using one receiver: application to microcrystalline and membrane protein preparations.

    PubMed

    Gopinath, T; Veglia, Gianluigi

    2015-04-01

    Solid-state NMR spectroscopy of proteins is a notoriously low-throughput technique. Relatively low-sensitivity and poor resolution of protein samples require long acquisition times for multidimensional NMR experiments. To speed up data acquisition, we developed a family of experiments called Polarization Optimized Experiments (POE), in which we utilized the orphan spin operators that are discarded in classical multidimensional NMR experiments, recovering them to allow simultaneous acquisition of multiple 2D and 3D experiments, all while using conventional probes with spectrometers equipped with one receiver. POE allow the concatenation of multiple 2D or 3D pulse sequences into a single experiment, thus potentially combining all of the aforementioned advances, boosting the capability of ssNMR spectrometers at least two-fold without the addition of any hardware. In this perspective, we describe the first generation of POE, such as dual acquisition MAS (or DUMAS) methods, and then illustrate the evolution of these experiments into MEIOSIS, a method that enables the simultaneous acquisition of multiple 2D and 3D spectra. Using these new pulse schemes for the solid-state NMR investigation of biopolymers makes it possible to obtain sequential resonance assignments, as well as distance restraints, in about half the experimental time. While designed for acquisition of heteronuclei, these new experiments can be easily implemented for proton detection and coupled with other recent advancements, such as dynamic nuclear polarization (DNP), to improve signal to noise. Finally, we illustrate the application of these methods to microcrystalline protein preparations as well as single and multi-span membrane proteins reconstituted in lipid membranes. PMID:25797011

  1. Multiple Acquisition of Magic Angle Spinning Solid-State NMR Experiments Using One Receiver: Application to Microcrystalline and Membrane Protein Preparations

    PubMed Central

    Gopinath, T.; Veglia, Gianluigi

    2015-01-01

    Solid-State NMR spectroscopy of proteins is a notoriously low-throughput technique. Relatively low-sensitivity and poor resolution of protein samples require long acquisition times for multidimensional NMR experiments. To speed up data acquisition, we developed a family of experiments called Polarization Optimized Experiments (POE), in which we utilized the orphan spin operators that are discarded in classical multidimensional NMR experiments, recovering them to allow simultaneous acquisition of multiple 2D and 3D experiments, all while using conventional probes with spectrometers equipped with one receiver. POEs allow the concatenation of multiple 2D or 3D pulse sequences into a single experiment, thus potentially combining all of the aforementioned advances, boosting the capability of ssNMR spectrometers at least two-fold without the addition of any hardware. In this Perspective, we describe the first generation of POEs, such as dual acquisition MAS (or DUMAS) methods, and then illustrate the evolution of these experiments into MEIOSIS, a method that enables the simultaneous acquisition of multiple 2D and 3D spectra. Using these new pulse schemes for the solid-state NMR investigation of biopolymers makes it possible to obtain sequential resonance assignments, as well as distance restraints, in about half the experimental time. While designed for acquisition of heteronuclei, these new experiments can be easily implemented for proton detection and coupled with other recent advancements, such as dynamic polarization, to improve signal to noise. Finally, we illustrate the application of these methods to microcrystalline protein preparations as well as single and multi-span membrane proteins reconstituted in lipid membranes. PMID:25797011

  2. Raman Spectroscopy and Solid State NMR Characterization of Carbonates and Metal-Oxide Clusters

    NASA Astrophysics Data System (ADS)

    Moore, J. K.; Hammann, B. A.; Surface, J. A.; Ma, Z. L.; Hayes, S. E.

    2014-06-01

    Presented here is data showing the utility of combining Raman spectroscopy and solid-state Nuclear Magnetic Resonance. These techniques were combined to study local structure of carbonate minerals and metal oxides.

  3. Solid-State 2H NMR Shows Equivalence of Dehydration and Osmotic Pressures in Lipid Membrane Deformation

    PubMed Central

    Mallikarjunaiah, K.J.; Leftin, Avigdor; Kinnun, Jacob J.; Justice, Matthew J.; Rogozea, Adriana L.; Petrache, Horia I.; Brown, Michael F.

    2011-01-01

    Lipid bilayers represent a fascinating class of biomaterials whose properties are altered by changes in pressure or temperature. Functions of cellular membranes can be affected by nonspecific lipid-protein interactions that depend on bilayer material properties. Here we address the changes in lipid bilayer structure induced by external pressure. Solid-state 2H NMR spectroscopy of phospholipid bilayers under osmotic stress allows structural fluctuations and deformation of membranes to be investigated. We highlight the results from NMR experiments utilizing pressure-based force techniques that control membrane structure and tension. Our 2H NMR results using both dehydration pressure (low water activity) and osmotic pressure (poly(ethylene glycol) as osmolyte) show that the segmental order parameters (SCD) of DMPC approach very large values of ≈0.35 in the liquid-crystalline state. The two stresses are thermodynamically equivalent, because the change in chemical potential when transferring water from the interlamellar space to the bulk water phase corresponds to the induced pressure. This theoretical equivalence is experimentally revealed by considering the solid-state 2H NMR spectrometer as a virtual osmometer. Moreover, we extend this approach to include the correspondence between osmotic pressure and hydrostatic pressure. Our results establish the magnitude of the pressures that lead to significant bilayer deformation including changes in area per lipid and volumetric bilayer thickness. We find that appreciable bilayer structural changes occur with osmotic pressures in the range of 10−100 atm or lower. This research demonstrates the applicability of solid-state 2H NMR spectroscopy together with bilayer stress techniques for investigating the mechanism of pressure sensitivity of membrane proteins. PMID:21190661

  4. Powder-XRD and (14) N magic angle-spinning solid-state NMR spectroscopy of some metal nitrides.

    PubMed

    Kempgens, Pierre; Britton, Jonathan

    2016-05-01

    Some metal nitrides (TiN, ZrN, InN, GaN, Ca3 N2 , Mg3 N2 , and Ge3 N4 ) have been studied by powder X-ray diffraction (XRD) and (14) N magic angle-spinning (MAS) solid-state NMR spectroscopy. For Ca3 N2 , Mg3 N2 , and Ge3 N4 , no (14) N NMR signal was observed. Low speed (νr  = 2 kHz for TiN, ZrN, and GaN; νr  = 1 kHz for InN) and 'high speed' (νr  = 15 kHz for TiN; νr  = 5 kHz for ZrN; νr  = 10 kHz for InN and GaN) MAS NMR experiments were performed. For TiN, ZrN, InN, and GaN, powder-XRD was used to identify the phases present in each sample. The number of peaks observed for each sample in their (14) N MAS solid-state NMR spectrum matches perfectly well with the number of nitrogen-containing phases identified by powder-XRD. The (14) N MAS solid-state NMR spectra are symmetric and dominated by the quadrupolar interaction. The envelopes of the spinning sidebands manifold are Lorentzian, and it is concluded that there is a distribution of the quadrupolar coupling constants Qcc 's arising from structural defects in the compounds studied. Copyright © 2015 John Wiley & Sons, Ltd. PMID:26687421

  5. 250 GHz CW Gyrotron Oscillator for Dynamic Nuclear Polarization in Biological Solid State NMR

    PubMed Central

    Bajaj, Vikram S.; Hornstein, Melissa K.; Kreischer, Kenneth E.; Sirigiri, Jagadishwar R.; Woskov, Paul P.; Mak-Jurkauskas, Melody L.; Herzfeld, Judith; Temkin, Richard J.; Griffin, Robert G.

    2009-01-01

    In this paper, we describe a 250 GHz gyrotron oscillator, a critical component of an integrated system for magic angle spinning (MAS) dynamic nuclear polarization (DNP) experiments at 9T, corresponding to 380 MHz 1H frequency. The 250 GHz gyrotron is the first gyro-device designed with the goal of seamless integration with an NMR spectrometer for routine DNP-enhanced NMR spectroscopy and has operated under computer control for periods of up to 21 days with a 100% duty cycle. Following a brief historical review of the field, we present studies of the membrane protein bacteriorhodopsin (bR) using DNP-enhanced multidimensional NMR. These results include assignment of active site resonances in [U-13C,15N]-bR and demonstrate the utility of DNP for studies of membrane proteins. Next, we review the theory of gyro-devices from quantum mechanical and classical viewpoints and discuss the unique considerations that apply to gyrotron oscillators designed for DNP experiments. We then characterize the operation of the 250 GHz gyrotron in detail, including its long-term stability and controllability. We have measured the spectral purity of the gyrotron emission using both homodyne and heterodyne techniques. Radiation intensity patterns from the corrugated waveguide that delivers power to the NMR probe were measured using two new techniques to confirm pure mode content: a thermometric approach based on the temperature-dependent color of liquid crystalline media applied to a substrate and imaging with a pyroelectric camera. We next present a detailed study of the mode excitation characteristics of the gyrotron. Exploration of the operating characteristics of several fundamental modes reveals broadband continuous frequency tuning of up to 1.8 GHz as a function of the magnetic field alone, a feature that may be exploited in future tunable gyrotron designs. Oscillation of the 250 GHz gyrotron at the second harmonic of cyclotron resonance begins at extremely low beam currents (as low 12 mA) at frequencies between 320–365 GHz, suggesting an efficient route for the generation of even higher frequency radiation. The low starting currents were attributed to an elevated cavity Q, which is confirmed by cavity thermal load measurements. We conclude with an appendix containing a detailed description of the control system that safely automates all aspects of the gyrotron operation. PMID:17942352

  6. 250GHz CW gyrotron oscillator for dynamic nuclear polarization in biological solid state NMR.

    PubMed

    Bajaj, Vikram S; Hornstein, Melissa K; Kreischer, Kenneth E; Sirigiri, Jagadishwar R; Woskov, Paul P; Mak-Jurkauskas, Melody L; Herzfeld, Judith; Temkin, Richard J; Griffin, Robert G

    2007-12-01

    In this paper, we describe a 250 GHz gyrotron oscillator, a critical component of an integrated system for magic angle spinning (MAS) dynamic nuclear polarization (DNP) experiments at 9T, corresponding to 380 MHz (1)H frequency. The 250 GHz gyrotron is the first gyro-device designed with the goal of seamless integration with an NMR spectrometer for routine DNP enhanced NMR spectroscopy and has operated under computer control for periods of up to 21 days with a 100% duty cycle. Following a brief historical review of the field, we present studies of the membrane protein bacteriorhodopsin (bR) using DNP enhanced multidimensional NMR. These results include assignment of active site resonances in [U-(13)C, (15)N]-bR and demonstrate the utility of DNP for studies of membrane proteins. Next, we review the theory of gyro-devices from quantum mechanical and classical viewpoints and discuss the unique considerations that apply to gyrotron oscillators designed for DNP experiments. We then characterize the operation of the 250 GHz gyrotron in detail, including its long-term stability and controllability. We have measured the spectral purity of the gyrotron emission using both homodyne and heterodyne techniques. Radiation intensity patterns from the corrugated waveguide that delivers power to the NMR probe were measured using two new techniques to confirm pure mode content: a thermometric approach based on the temperature-dependent color of liquid crystalline media applied to a substrate and imaging with a pyroelectric camera. We next present a detailed study of the mode excitation characteristics of the gyrotron. Exploration of the operating characteristics of several fundamental modes reveals broadband continuous frequency tuning of up to 1.8 GHz as a function of the magnetic field alone, a feature that may be exploited in future tunable gyrotron designs. Oscillation of the 250 GHz gyrotron at the second harmonic of cyclotron resonance begins at extremely low beam currents (as low 12 mA) at frequencies between 320 and 365 GHz, suggesting an efficient route for the generation of even higher frequency radiation. The low starting currents were attributed to an elevated cavity Q, which is confirmed by cavity thermal load measurements. We conclude with an appendix containing a detailed description of the control system that safely automates all aspects of the gyrotron operation. PMID:17942352

  7. Efficient design of multituned transmission line NMR probes: the electrical engineering approach.

    PubMed

    Frydel, J A; Krzystyniak, M; Pienkowski, D; Pietrzak, M; de Sousa Amadeu, N; Ratajczyk, T; Idzik, K; Gutmann, T; Tietze, D; Voigt, S; Fenn, A; Limbach, H H; Buntkowsky, G

    2011-01-01

    Transmission line-based multi-channel solid state NMR probes have many advantages regarding the cost of construction, number of RF-channels, and achievable RF-power levels. Nevertheless, these probes are only rarely employed in solid state-NMR-labs, mainly owing to the difficult experimental determination of the necessary RF-parameters. Here, the efficient design of multi-channel solid state MAS-NMR probes employing transmission line theory and modern techniques of electrical engineering is presented. As technical realization a five-channel ((1)H, (31)P, (13)C, (2)H and (15)N) probe for operation at 7 Tesla is described. This very cost efficient design goal is a multi port single coil transmission line probe based on the design developed by Schaefer and McKay. The electrical performance of the probe is determined by measuring of Scattering matrix parameters (S-parameters) in particular input/output ports. These parameters are compared to the calculated parameters of the design employing the S-matrix formalism. It is shown that the S-matrix formalism provides an excellent tool for examination of transmission line probes and thus the tool for a rational design of these probes. On the other hand, the resulting design provides excellent electrical performance. From a point of view of Nuclear Magnetic Resonance (NMR), calibration spectra of particular ports (channels) are of great importance. The estimation of the π/2 pulses length for all five NMR channels is presented. PMID:21316931

  8. Tannin fingerprinting in vegetable tanned leather by solid state NMR spectroscopy and comparison with leathers tanned by other processes.

    PubMed

    Romer, Frederik H; Underwood, Andrew P; Senekal, Nadine D; Bonnet, Susan L; Duer, Melinda J; Reid, David G; van der Westhuizen, Jan H

    2011-01-01

    Solid state ¹³C-NMR spectra of pure tannin powders from four different sources--mimosa, quebracho, chestnut and tara--are readily distinguishable from each other, both in pure commercial powder form, and in leather which they have been used to tan. Groups of signals indicative of the source, and type (condensed vs. hydrolyzable) of tannin used in the manufacture are well resolved in the spectra of the finished leathers. These fingerprints are compared with those arising from leathers tanned with other common tanning agents. Paramagnetic chromium (III) tanning causes widespread but selective disappearance of signals from the spectrum of leather collagen, including resonances from acidic aspartyl and glutamyl residues, likely bound to Cr (III) structures. Aluminium (III) and glutaraldehyde tanning both cause considerable leather collagen signal sharpening suggesting some increase in molecular structural ordering. The ²⁷Al-NMR signal from the former material is consistent with an octahedral coordination by oxygen ligands. Solid state NMR thus provides easily recognisable reagent specific spectral fingerprints of the products of vegetable and some other common tanning processes. Because spectra are related to molecular properties, NMR is potentially a powerful tool in leather process enhancement and quality or provenance assurance. PMID:21278677

  9. Synthesis and conformational study of silk model polypeptides [Ala-Gly] 12 by solid-state NMR

    NASA Astrophysics Data System (ADS)

    Kishi, Satoshi; Santos, Andres; Ishii, Osamu; Ishikawa, Kazuo; Kunieda, Shingo; Kimura, Hideaki; Shoji, Akira

    2003-04-01

    A series of well-defined model polypeptides of Bombyx mori silk fibroin, [Ala-Gly] n ( n=12 and 5-9), [Ala d4Gly] 12 and [Gly d2Ala] 12, (where Ala, Gly, Ala d4 and Gly d2 denote L-alanine, glycine, 2,3,3,3-deuterated L-alanine and 2,2-deuterated glycine residues, respectively), were successfully synthesized by an automatic solid phase peptide synthesizer. Followed by treatment with 9 M lithium bromide (LiBr) solution to convert their conformation from the silk II to the silk I form. We determined the intrinsic 1H, 13C and 15N NMR chemical shifts of Ala and Gly residues, characteristic to the silk I and silk II forms in Bombyx mori, by high-resolution and solid-state 13C CP-MAS, 1H CRAMPS, 1H- 13C HETCOR and 15N CP-MAS NMR measurements. Next, we demonstrated that it is necessary at least 12 amino acid residues ( n=6) in the [Ala-Gly] n-I series for the formation of stable silk I form by 13C CP-MAS NMR. Finally, we found that the intrinsic 1H and 13C chemical shifts of the specific deuterium labeled polypeptides, [Ala d4-Gly] 12 and [Gly d2-Ala] 12, cause a meaningful chemical shift displacement, suggesting that the deuterium interacts with proton and/or carbon atoms in the solid state. Thus, the selective deuterium labeling has a potential to useful for the structural analysis by 1H NMR in the solid state.

  10. Quantitative structural constraints for organic powders at natural isotopic abundance using dynamic nuclear polarization solid-state NMR spectroscopy.

    PubMed

    Mollica, Giulia; Dekhil, Myriam; Ziarelli, Fabio; Thureau, Pierre; Viel, Stéphane

    2015-05-11

    A straightforward method is reported to quantitatively relate structural constraints based on (13)C-(13)C double-quantum build-up curves obtained by dynamic nuclear polarization (DNP) solid-state NMR to the crystal structure of organic powders at natural isotopic abundance. This method relies on the significant gain in NMR sensitivity provided by DNP (approximately 50-fold, lowering the experimental time from a few years to a few days), and is sensitive to the molecular conformation and crystal packing of the studied powder sample (in this case theophylline). This method allows trial crystal structures to be rapidly and effectively discriminated, and paves the way to three-dimensional structure elucidation of powders through combination with powder X-ray diffraction, crystal-structure prediction, and density functional theory computation of NMR chemical shifts. PMID:25809550

  11. Characterization of Al30 in commercial poly-aluminum chlorohydrate by solid-state (27)Al NMR spectroscopy.

    PubMed

    Phillips, Brian L; Vaughn, John S; Smart, Scott; Pan, Long

    2016-08-15

    Investigation of commercially produced hydrolysis salts of aluminum by solid-state (27)Al NMR spectroscopy and size-exclusion chromatography (SEC) reveals well-defined and distinct Al environments that can be related to physicochemical properties. (27)Al MAS and MQ-MAS NMR spectroscopic data show that the local structure of the solids is dominated by moieties that closely resemble the Al30 polyoxocation (Al30O8(OH)56(H2O)26(18+)), accounting for 72-85% of the total Al. These Al30-like clusters elute as several size fractions by SEC. Comparison of the SEC and NMR results indicates that the Al30-like clusters includes intact isolated clusters, moieties of larger polymers or aggregates, and possibly fragments resembling δ-Al13 Keggin clusters. The coagulation efficacy of the solids appears to correlate best with the abundance of intact Al30-like clusters and of smaller species available to promote condensation reactions. PMID:27232539

  12. Towards structure determination of neurotoxin II bound to nicotinic acetylcholine receptor: a solid-state NMR approach.

    PubMed

    Krabben, Ludwig; van Rossum, Barth-Jan; Castellani, Federica; Bocharov, Eduard; Schulga, Alexey A; Arseniev, Alexander S; Weise, Christoph; Hucho, Ferdinand; Oschkinat, Hartmut

    2004-04-30

    Solid-state magic-angle spinning nuclear magnetic resonance (NMR) has sufficient resolving power for full assignment of resonances and structure determination of immobilised biological samples as was recently shown for a small microcrystalline protein. In this work, we show that highly resolved spectra may be obtained from a system composed of a receptor-toxin complex. The NMR sample used for our studies consists of a membrane preparation of the nicotinic acetylcholine receptor from the electric organ of Torpedo californica which was incubated with uniformly 13C-,15N-labelled neurotoxin II. Despite the large size of the ligand-receptor complex ( > 290 kDa) and the high lipid content of the sample, we were able to detect and identify residues from the ligand. The comparison with solution NMR data of the free toxin indicates that its overall structure is very similar when bound to the receptor, but significant changes were observed for one isoleucine. PMID:15111116

  13. A high-resolution sup 13 C solid-state NMR study of meso-tetraphenylporphyrin and its zinc(II) complex

    SciTech Connect

    Rocha, J.; Kolodziejski, W.; Klinowski, J. ); Cavaleiro, J.A.S. )

    1992-01-01

    High-resolution {sup 13}C solid-state NMR spectra of meso-tetraphenylporphyrin (TPP) and its zinc(II) complex (ZnTPP) are assigned by reference to low-temperature solution NMR results and using {sup 1}H- {sup 13}C cross-polarization magic-angle-spinning (CP/MAS). The splittings of the signals from pyrrole carbons in TPP are attributed to kinetic solid-state states involved in the migration of the central hydrogen atom.

  14. Chemical and nanometer-scale structure of kerogen and its change during thermal maturation investigated by advanced solid-state 13C NMR spectroscopy

    USGS Publications Warehouse

    Mao, J.; Fang, X.; Lan, Y.; Schimmelmann, A.; Mastalerz, Maria; Xu, L.; Schmidt-Rohr, K.

    2010-01-01

    We have used advanced and quantitative solid-state nuclear magnetic resonance (NMR) techniques to investigate structural changes in a series of type II kerogen samples from the New Albany Shale across a range of maturity (vitrinite reflectance R0 from 0.29% to 1.27%). Specific functional groups such as CH3, CH2, alkyl CH, aromatic CH, aromatic C-O, and other nonprotonated aromatics, as well as "oil prone" and "gas prone" carbons, have been quantified by 13C NMR; atomic H/C and O/C ratios calculated from the NMR data agree with elemental analysis. Relationships between NMR structural parameters and vitrinite reflectance, a proxy for thermal maturity, were evaluated. The aromatic cluster size is probed in terms of the fraction of aromatic carbons that are protonated (???30%) and the average distance of aromatic C from the nearest protons in long-range H-C dephasing, both of which do not increase much with maturation, in spite of a great increase in aromaticity. The aromatic clusters in the most mature sample consist of ???30 carbons, and of ???20 carbons in the least mature samples. Proof of many links between alkyl chains and aromatic rings is provided by short-range and long-range 1H-13C correlation NMR. The alkyl segments provide most H in the samples; even at a carbon aromaticity of 83%, the fraction of aromatic H is only 38%. While aromaticity increases with thermal maturity, most other NMR structural parameters, including the aromatic C-O fractions, decrease. Aromaticity is confirmed as an excellent NMR structural parameter for assessing thermal maturity. In this series of samples, thermal maturation mostly increases aromaticity by reducing the length of the alkyl chains attached to the aromatic cores, not by pronounced growth of the size of the fused aromatic ring clusters. ?? 2010 Elsevier Ltd. All rights reserved.

  15. Multiple acquisitions via sequential transfer of orphan spin polarization (MAeSTOSO): How far can we push residual spin polarization in solid-state NMR?

    PubMed

    Gopinath, T; Veglia, Gianluigi

    2016-06-01

    Conventional multidimensional magic angle spinning (MAS) solid-state NMR (ssNMR) experiments detect the signal arising from the decay of a single coherence transfer pathway (FID), resulting in one spectrum per acquisition time. Recently, we introduced two new strategies, namely DUMAS (DUal acquisition Magic Angle Spinning) and MEIOSIS (Multiple ExperIments via Orphan SpIn operatorS), that enable the simultaneous acquisitions of multidimensional ssNMR experiments using multiple coherence transfer pathways. Here, we combined the main elements of DUMAS and MEIOSIS to harness both orphan spin operators and residual polarization and increase the number of simultaneous acquisitions. We show that it is possible to acquire up to eight two-dimensional experiments using four acquisition periods per each scan. This new suite of pulse sequences, called MAeSTOSO for Multiple Acquisitions via Sequential Transfer of Orphan Spin pOlarization, relies on residual polarization of both (13)C and (15)N pathways and combines low- and high-sensitivity experiments into a single pulse sequence using one receiver and commercial ssNMR probes. The acquisition of multiple experiments does not affect the sensitivity of the main experiment; rather it recovers the lost coherences that are discarded, resulting in a significant gain in experimental time. Both merits and limitations of this approach are discussed. PMID:27039168

  16. Solid-state NMR studies for DC 745: April--May 2000

    SciTech Connect

    Stephens, T.; Labouriau, A.

    2000-12-01

    Powles et al. reported in 1961 that pdydimethyl siloxane melts exhibit a transverse decay of nuclear spin magnetization (T{sub 2}) that is nonexponential. This observation was interpreted in terms of the proton NMR signal from the melt having characteristics of a broad line. Since then, similar findings were obtained for other polymer melts too. Accordingly to Powles et al. this peculiar proton line shape was the result of an intramolecular direct dipole-dipole interaction between a pair of protons. They also suggested that this dipolar interaction was weak because the dipolar interaction was motionally averaged since the orientation-dependent local dipolar field is modulated by molecular dynamics. In sum, the consequences and the origins of nuclear spin correlations in viscous polymer melts were recognized almost 4 decades ago. In the 1970s, Cohen-Addad suggested that these weak proton dipolar interactions could be used to characterize polymer melts and networks. In this work, we have used some of these ideas to probe the effects of thermal treatments on the proton spin-spin relaxation times for DC745.

  17. Acidic Properties and Structure-Activity Correlations of Solid Acid Catalysts Revealed by Solid-State NMR Spectroscopy.

    PubMed

    Zheng, Anmin; Li, Shenhui; Liu, Shang-Bin; Deng, Feng

    2016-04-19

    Solid acid materials with tunable structural and acidic properties are promising heterogeneous catalysts for manipulating and/or emulating the activity and selectivity of industrially important catalytic reactions. On the other hand, the performances of acid-catalyzed reactions are mostly dictated by the acidic features, namely, type (Brønsted vs Lewis acidity), amount, strength, and local environment of acid sites. The latter is relevant to their location (intra- vs extracrystalline), and possible confinement and Brønsted-Lewis acid synergy effects that may strongly affect the host-guest interactions, reaction mechanism, and shape selectivity of the catalytic system. This account aims to highlight some important applications of state-of-the-art solid-state NMR (SSNMR) techniques for exploring the structural and acidic properties of solid acid catalysts as well as their catalytic performances and relevant reaction pathway invoked. In addition, density functional theory (DFT) calculations may be exploited in conjunction with experimental SSNMR studies to verify the structure-activity correlations of the catalytic system at a microscopic scale. We describe in this Account the developments and applications of advanced ex situ and/or in situ SSNMR techniques, such as two-dimensional (2D) double-quantum magic-angle spinning (DQ MAS) homonuclear correlation spectroscopy for structural investigation of solid acids as well as study of their acidic properties. Moreover, the energies and electronic structures of the catalysts and detailed catalytic reaction processes, including the identification of reaction species, elucidation of reaction mechanism, and verification of structure-activity correlations, made available by DFT theoretical calculations were also discussed. Relevant discussions will focus primarily on results obtained from our laboratories in the past decade, including (i) quantitative and qualitative acidity characterization utilizing assorted probe molecules, (ii) probing the spatial proximity and synergy effect of acid sites, and (iii) influence of acid features and pore confinement effect on catalytic activity, transition-state stability, reaction pathway, and product selectivity of solid acid catalysts such as zeolites, metal oxides, and heteropolyacids. It is conclusive that a synergy of acidity (local effect) and pore confinement (environmental effect) tend to strongly dictate the formations of intermediates and transition states, hence, the reaction pathways and catalytic performance of solid acid catalysts. We hope that these information can provide additional insights toward our understanding in heterogeneous catalysis, especially the roles of structural and acidic properties on catalytic performances and reaction mechanism of acid-catalyzed systems, which should be beneficial for rational design of solid acid catalysts. PMID:26990961

  18. Solid-state 2H MAS NMR studies of TNT absorption in soil and clays.

    PubMed

    Emery, E F; Junk, T; Ferrell, R E; De Hon, R; Butler, L G

    2001-07-15

    The binding of TNT to a soil and soil components is studied with 2H MAS NMR. This NMR technique clearly shows different interactions for deuterated TNT with soil, quartz, and clays. Basically, this NMR method relies on the well-defined 2H MAS NMR characteristics of a deuterated methyl group attached to an aromatic ring. Stationary rings and rings executing 2- or 3-fold jump motions all yield different NMR spectra. The technique has good sensitivity; these studies were conducted at a TNT loading of 22 mmol/kg. For the soil sample studied, TNT showed binding similar to a TNT/K10-montmorillonite sample. PMID:11478251

  19. EASY-GOING deconvolution: Combining accurate simulation and evolutionary algorithms for fast deconvolution of solid-state quadrupolar NMR spectra

    NASA Astrophysics Data System (ADS)

    Grimminck, Dennis L. A. G.; Polman, Ben J. W.; Kentgens, Arno P. M.; Leo Meerts, W.

    2011-08-01

    A fast and accurate fit program is presented for deconvolution of one-dimensional solid-state quadrupolar NMR spectra of powdered materials. Computational costs of the synthesis of theoretical spectra are reduced by the use of libraries containing simulated time/frequency domain data. These libraries are calculated once and with the use of second-party simulation software readily available in the NMR community, to ensure a maximum flexibility and accuracy with respect to experimental conditions. EASY-GOING deconvolution ( EGdeconv) is equipped with evolutionary algorithms that provide robust many-parameter fitting and offers efficient parallellised computing. The program supports quantification of relative chemical site abundances and (dis)order in the solid-state by incorporation of (extended) Czjzek and order parameter models. To illustrate EGdeconv's current capabilities, we provide three case studies. Given the program's simple concept it allows a straightforward extension to include other NMR interactions. The program is available as is for 64-bit Linux operating systems.

  20. Bottom-up and top-down solid-state NMR approaches for bacterial biofilm matrix composition

    NASA Astrophysics Data System (ADS)

    Cegelski, Lynette

    2015-04-01

    The genomics and proteomics revolutions have been enormously successful in providing crucial "parts lists" for biological systems. Yet, formidable challenges exist in generating complete descriptions of how the parts function and assemble into macromolecular complexes and whole-cell assemblies. Bacterial biofilms are complex multicellular bacterial communities protected by a slime-like extracellular matrix that confers protection to environmental stress and enhances resistance to antibiotics and host defenses. As a non-crystalline, insoluble, heterogeneous assembly, the biofilm extracellular matrix poses a challenge to compositional analysis by conventional methods. In this perspective, bottom-up and top-down solid-state NMR approaches are described for defining chemical composition in complex macrosystems. The "sum-of-the-parts" bottom-up approach was introduced to examine the amyloid-integrated biofilms formed by Escherichia coli and permitted the first determination of the composition of the intact extracellular matrix from a bacterial biofilm. An alternative top-down approach was developed to define composition in Vibrio cholerae biofilms and relied on an extensive panel of NMR measurements to tease out specific carbon pools from a single sample of the intact extracellular matrix. These two approaches are widely applicable to other heterogeneous assemblies. For bacterial biofilms, quantitative parameters of matrix composition are needed to understand how biofilms are assembled, to improve the development of biofilm inhibitors, and to dissect inhibitor modes of action. Solid-state NMR approaches will also be invaluable in obtaining parameters of matrix architecture.

  1. Solid-State (17)O NMR of Oxygen-Nitrogen Singly Bonded Compounds: Hydroxylammonium Chloride and Sodium Trioxodinitrate (Angeli's Salt).

    PubMed

    Lu, Jiasheng; Kong, Xianqi; Terskikh, Victor; Wu, Gang

    2015-07-23

    We report a solid-state NMR study of (17)O-labeled hydroxylammonium chloride ([H(17)O-NH3]Cl) and sodium trioxodinitrate monohydrate (Na2[(17)ONNO2]·H2O, Angeli's salt). The common feature in these two compounds is that they both contain oxygen atoms that are singly bonded to nitrogen. For this class of oxygen-containing functional groups, there is very limited solid-state (17)O NMR data in the literature. In this work, we experimentally measured the (17)O chemical shift and quadrupolar coupling tensors. With the aid of plane-wave DFT computation, the (17)O NMR tensor orientations were determined in the molecular frame of reference. We found that the characteristic feature of an O-N single bond is that the (17)O nucleus exhibits a large quadrupolar coupling constant (13-15 MHz) but a rather small chemical shift anisotropy (100-250 ppm), in sharp contrast with the nitroso (O═N) functional group for which both quantities are very large (e.g., 16 MHz and 3000 ppm, respectively). PMID:26107984

  2. Protein secondary structure of Green Lynx spider dragline silk investigated by solid-state NMR and X-ray diffraction.

    PubMed

    Xu, Dian; Shi, Xiangyan; Thompson, Forrest; Weber, Warner S; Mou, Qiushi; Yarger, Jeffery L

    2015-11-01

    In this study, the secondary structure of the major ampullate silk from Peucetia viridans (Green Lynx) spiders is characterized by X-ray diffraction and solid-state NMR spectroscopy. From X-ray diffraction measurement, β-sheet nanocrystallites were observed and found to be highly oriented along the fiber axis, with an orientational order, fc≈0.98. The size of the nanocrystallites was determined to be on average 2.5nm×3.3nm×3.8nm. Besides a prominent nanocrystalline region, a partially oriented amorphous region was also observed with an fa≈0.89. Two-dimensional (13)C-(13)C through-space and through-bond solid-state NMR experiments were employed to elucidate structure details of P. viridans silk proteins. It reveals that β-sheet nanocrystallites constitutes 40.0±1.2% of the protein and are dominated by alanine-rich repetitive motifs. Furthermore, based upon the NMR data, 18±1% of alanine, 60±2% glycine and 54±2% serine are incorporated into helical conformations. PMID:26226457

  3. Detection of fulleroid sites in fullerene-60 by high-resolution solid-state [sup 1]H NMR

    SciTech Connect

    Kolodziejski, W.; Corma, A. ); Barras, J.; Klinowski, J. )

    1995-03-09

    [sup 1]H solid-state NMR with magic-angle spinning (MAS NMR) detects dilute H-containing species in chromatographically purified C[sub 60]. Because of the weakness of the [sup 1]H-[sup 1]H dipolar couplings, the spectral peaks are very sharp and the T[sub 1] values are not averaged by spin diffusion. On the basis of chemical shifts and peak connectivities as revealed by COSY, we have identified various fulleroids, as well as toluene and dioctyl phthalate (DOP) impurities. The fulleroids contain HC=CHCH[sub 2]CH[sub 3], HC=CHCH[sub 3], and HC=CH groups. DOP is probably complexed to C[sub 60] via the aromatic moiety. Laboratory and rotating-frame spin-diffusion experiments show that the H-containing species are so diluted by bulk C[sub 60] that they cannot communicate by intermolecular spin diffusion. The intramolecular spin-diffusion paths suggest that the alkyl chains of DOP are folded back toward the aromatic ring. We have found that two-dimensional [sup 1]H NMR techniques work well in the solid state without a multiple-pulse homonuclear decoupling, provided that the species under study are sufficiently dilute by a nondipolar medium. 46 refs., 8 figs., 2 tabs.

  4. Bottom-Up and Top-Down Solid-State NMR Approaches for Bacterial Biofilm Matrix Composition

    PubMed Central

    Cegelski, Lynette

    2015-01-01

    The genomics and proteomics revolutions have been enormously successful in providing crucial “parts lists” for biological systems. Yet, formidable challenges exist in generating complete descriptions of how the parts function and assemble into macromolecular complexes and whole-cell assemblies. Bacterial biofilms are complex multicellular bacterial communities protected by a slime-like extracellular matrix that confers protection to environmental stress and enhances resistance to antibiotics and host defenses. As a non-crystalline, insoluble, heterogeneous assembly, the biofilm extracellular matrix poses a challenge to compositional analysis by conventional methods. In this Perspective, bottom-up and top-down solid-state NMR approaches are described for defining chemical composition in complex macrosystems. The “sum-of-theparts” bottom-up approach was introduced to examine the amyloid-integrated biofilms formed by E. coli and permitted the first determination of the composition of the intact extracellular matrix from a bacterial biofilm. An alternative top-down approach was developed to define composition in V. cholerae biofilms and relied on an extensive panel of NMR measurements to tease out specific carbon pools from a single sample of the intact extracellular matrix. These two approaches are widely applicable to other heterogeneous assemblies. For bacterial biofilms, quantitative parameters of matrix composition are needed to understand how biofilms are assembled, to improve the development of biofilm inhibitors, and to dissect inhibitor modes of action. Solid-state NMR approaches will also be invaluable in obtaining parameters of matrix architecture. PMID:25797008

  5. Solid state {sup 31}P NMR study of phosphonate binding sites in guanidine-functionalized, molecular imprinted silica xerogels

    SciTech Connect

    Sasaki, D.Y.; Alam, T.D.

    2000-01-03

    Phosphonate binding sites in guanidine and ammonium surface-functionalized silica xerogels were prepared via the molecular imprinting technique and characterized using solid state {sup 31}P MAS NMR. One-point, two-point, and non-specific host-guest interactions between phenylphosphonic acid (PPA) and the functionalized gels were distinguished by characteristic chemical shifts of the observed absorption peaks. Using solid state as well as solution phase NMR analyses, absorptions observed at 15.5 ppm and 6.5 ppm were identified as resulting from the 1:1 (one-point) and 2:1 (two-point) guanidine to phosphonate interactions, respectively. Similar absorptions were observed with the ammonium functionalized gels. By examining the host-guest interactions within the gels, the efficiency of the molecular imprinting procedure with regard to the functional monomer-to-template interaction could be readily assessed. Template removal followed by substrate adsorption studies conducted on the guanidine functionalized gels provided a method to evaluate the binding characteristics of the receptor sites to a phosphonate substrate. During these experiments, {sup 29}Si and {sup 31}P MAS NMR acted as diagnostic monitors to identify structural changes occurring in the gel matrix and at the receptor site from solvent mediated processes.

  6. Structural analysis of a banana-liquid crystal in the B4 phase by solid-state NMR.

    PubMed

    Yamada, Kazuhiko; Kang, Sungmin; Takimoto, Koji; Hattori, Masaya; Shirata, Kei; Kawauchi, Susumu; Deguchi, Kenzo; Shimizu, Tadashi; Watanabe, Junji

    2013-06-01

    In this paper, we present a structural investigation of 1,3-phenylene bis[4-((4-10-decyloxyphenyl)iminomethyl)-benzoate], known as a banana-liquid crystal, in the B4 phase, which was performed by solid-state nuclear magnetic resonance (NMR) methodology combined with quantum chemical calculations. The present solid-state NMR measurements including (13)C CPMAS, 2D TOSS-deTOSS, dipole-dephase, 1D and 2D EXSY, and MAS-j-HMQC provided accurate spectral assignments and unambiguous NMR parameters such as (13)C chemical shift tensors, which were used for construction of the three-dimensional structure with the aid of density functional theory calculations. In the obtained molecular structure, two arms of the bent-core molecule are asymmetrically expanded such that the direction of the dipole moment is off alignment with respect to the middle line of the center benzene ring. It is this antisymmetric structure that is the origin of the twisted helical system in the B4 phase. PMID:23654351

  7. Solid-state 13C NMR analysis of Lower Cretaceous Baganuur (Mongolia) lignite

    NASA Astrophysics Data System (ADS)

    Erdenetsogt, B.; Lee, I.; Lee, S.; Ko, Y.

    2009-12-01

    The transformation of plant matter into peat and coal has two steps, called the biochemical and geochemical stages of coalification. Biochemical coalification begins with the accumulation of dead vegetable matter and ends at the rank of subbituminous coal. The rank of Baganuur lignite ranges from lignite to subbituminous coal. It is transition between biochemical and physico-chemical coalification stages. The changes of chemical structure of coal during the transition between above mentioned two stages were studied by solid state CP/MAS 13C NMR. The most predominant alteration is the disappearance of the resonances from oxygenated aliphatic carbons (63 ppm), protonated aromatic carbons (114 ppm), oxygen-substituted aromatic carbons (144 ppm) and carbonyl carbons (195 ppm). In addition, the intensity of resonances from methoxyl carbons (56 ppm) and oxygenated aliphatic carbons (72 ppm) decreased. While the intensities of resonance from aliphatic (30 ppm), protonated aromatic (125 ppm) and carboxyl carbon (174 ppm) increased or remained almost constant. The relative percent of O-substituted aromatic carbons decreased by ~25% mainly due to the intensity loss of the peak at 144 ppm, indicating removal of O-containing functional groups substituted to aromatic carbons. It is consistent with the decreased relative percent (~75%) of the peak at 114 ppm from protonated aromatic carbons nearby oxygen-substituted aromatic carbons. In addition, the resonance from 125 ppm was shifted to 128 ppm and its relative area increased by ~20%, indicating replacement of O-substituent of aromatic rings by hydrogen or carbon. Protonated aromatic carbons at least two bond away from an oxygen-substituted aromatic carbons give a resonance at 125 ppm and carbon-substituted aromatic carbons give a resonance at 130-132 ppm. With the increase relative percent of C-substituted aromatic carbons, their resonance were overlapped with protonated aromatic carbons and shifted to higher ppm. A decreasing area of oxygenated aliphatic carbons (~15 %) could be explained by β-O-4 ether cleavage and loss of hydroxyl groups from side-chains as well as complete removal of cellulosic material. In addition, those deoxygenating reactions are more likely responsible for the increased relative intensity of aliphatic carbons. In detail, the relative percent of methyl groups at 14 ppm remained almost constant (decreased only by ~4%), while that of methylene increased by 20%. During the β-O-4 ether cleavage and loss of hydroxyl groups from side-chains, relative amount of CH2 should increase in respect to O-containing original structures. Finally, the relative percentage of carboxyl/carboxyl carbons were decreased by ~25%, mainly due to diminished intensity of carbonyl carbons at 195 ppm. The intensity loss of carbonyl carbons increased from biochemical stage (~10%) to the beginning of physico-chemical stage (~70%).

  8. Investigation of the interface in silica-encapsulated liposomes by combining solid state NMR and first principles calculations.

    PubMed

    Folliet, Nicolas; Roiland, Claire; Bégu, Sylvie; Aubert, Anne; Mineva, Tzonka; Goursot, Annick; Selvaraj, Kaliaperumal; Duma, Luminita; Tielens, Frederik; Mauri, Francesco; Laurent, Guillaume; Bonhomme, Christian; Gervais, Christel; Babonneau, Florence; Azaïs, Thierry

    2011-10-26

    In the context of nanomedicine, liposils (liposomes and silica) have a strong potential for drug storage and release schemes: such materials combine the intrinsic properties of liposome (encapsulation) and silica (increased rigidity, protective coating, pH degradability). In this work, an original approach combining solid state NMR, molecular dynamics, first principles geometry optimization, and NMR parameters calculation allows the building of a precise representation of the organic/inorganic interface in liposils. {(1)H-(29)Si}(1)H and {(1)H-(31)P}(1)H Double Cross-Polarization (CP) MAS NMR experiments were implemented in order to explore the proton chemical environments around the silica and the phospholipids, respectively. Using VASP (Vienna Ab Initio Simulation Package), DFT calculations including molecular dynamics, and geometry optimization lead to the determination of energetically favorable configurations of a DPPC (dipalmitoylphosphatidylcholine) headgroup adsorbed onto a hydroxylated silica surface that corresponds to a realistic model of an amorphous silica slab. These data combined with first principles NMR parameters calculations by GIPAW (Gauge Included Projected Augmented Wave) show that the phosphate moieties are not directly interacting with silanols. The stabilization of the interface is achieved through the presence of water molecules located in-between the head groups of the phospholipids and the silica surface forming an interfacial H-bonded water layer. A detailed study of the (31)P chemical shift anisotropy (CSA) parameters allows us to interpret the local dynamics of DPPC in liposils. Finally, the VASP/solid state NMR/GIPAW combined approach can be extended to a large variety of organic-inorganic hybrid interfaces. PMID:21899369

  9. A new approach to determining homopolymer domain sizes in polycarbonate-polyether dialysis membranes by solid-state NMR.

    PubMed

    Lewis, R H; Wiesner, U; Lehmann, S; Spiess, H W

    1994-01-01

    The size of micro-separated domains of polyether (PEO), -[CH2CH2O]n-, and polycarbonate (PC), -[(C6H4)-C(CH3)2-(C6H4)-OCO2]m-, in the dialysis membrane 'Gambrane' have been determined using an advanced solid-state NMR technique which exploits differences in 1H spin diffusion. The characteristic diameter of a PEO domain is 4.8 +/- 1.4 nm and that of PC is 5.2 +/- 1.4 nm with a mixed phase region of 0.8 +/- 0.5 nm. PMID:7873508

  10. Multiple Acquisition/Multiple Observation Separated Local Field/Chemical Shift Correlation Solid-state Magic Angle Spinning NMR Spectroscopy

    PubMed Central

    Das, Bibhuti B.; Opella, Stanley J.

    2014-01-01

    Multiple acquisition spectroscopy (MACSY) experiments that enable multiple free induction decays to be recorded during individual experiments are demonstrated. In particular, the experiments incorporate separated local field spectroscopy into homonuclear and heteronuclear correlation spectroscopy. The measured heteronuclear dipolar couplings are valuable in structure determination as well as in enhancing resolution by providing an additional frequency axis. In one example four different three-dimensional spectra are obtained in a single experiment, demonstrating that substantial potential saving in experimental time is available when multiple multi-dimensional spectra are required as part of solid-state NMR studies. PMID:25023566

  11. Crystallinity and compositional changes in carbonated apatites: Evidence from 31P solid-state NMR, Raman, and AFM analysis

    PubMed Central

    McElderry, John-David P.; Zhu, Peizhi; Mroue, Kamal H.; Xu, Jiadi; Pavan, Barbara; Fang, Ming; Zhao, Guisheng; McNerny, Erin; Kohn, David H.; Franceschi, Renny T.; Holl, Mark M. Banaszak; Tecklenburg, Mary M.J.; Ramamoorthy, Ayyalusamy; Morris, Michael D.

    2013-01-01

    Solid-state (magic-angle spinning) NMR spectroscopy is a useful tool for obtaining structural information on bone organic and mineral components and synthetic model minerals at the atomic-level. Raman and 31P NMR spectral parameters were investigated in a series of synthetic B-type carbonated apatites (CAps). Inverse 31P NMR linewidth and inverse Raman PO43− ν1 bandwidth were both correlated with powder XRD c-axis crystallinity over the 0.3–10.3 wt% CO32− range investigated. Comparison with bone powder crystallinities showed agreement with values predicted by NMR and Raman calibration curves. Carbonate content was divided into two domains by the 31P NMR chemical shift frequency and the Raman phosphate ν1 band position. These parameters remain stable except for an abrupt transition at 6.5 wt% carbonate, a composition which corresponds to an average of one carbonate per unit cell. This near-binary distribution of spectroscopic properties was also found in AFM-measured particle sizes and Ca/P molar ratios by elemental analysis. We propose that this transition differentiates between two charge-balancing ion-loss mechanisms as measured by Ca/P ratios. These results define a criterion for spectroscopic characterization of B-type carbonate substitution in apatitic minerals. PMID:24273344

  12. 11B solid-state NMR investigation of the rhamnogalacturonan II-borate complex in plant cell walls.

    PubMed

    Kameda, Tsunenori; Ishii, Tadashi; Matsunaga, Toshiro; Ashida, Jun

    2006-02-01

    The boron in plant cell walls, which is water-insoluble and in the solid state, is solubilized by pectinase digestion to give a dimeric rhamnogalacturonan II-borate (dRG-II-B) complex. To clarify the nondestructive structure of boron present in plant cell walls (as represented by sugar beet fiber), we performed 192- and 96-MHz 11B solid state NMR measurements. The use of a high field magnet frequency of 192-MHz enabled us to observe 11B isotropic chemical shifts at -9.7 and -9.6 ppm for dRG-II-B and sugar beet fiber in the solid state, respectively, demonstrating that the boron in isolated dRG-II-B and in plant cell walls is present as a borate-diol ester (1:2). The observation of the magnetic field dependence of the chemical shift and lineshape for the borate-diol ester (1:2) by quadrupolar interaction suggested that the borate complex had a distorted tetrahedral boron structure. PMID:16512431

  13. Some studies on a solid-state sulfur probe for coal gasification systems

    NASA Technical Reports Server (NTRS)

    Jacob, K. T.; Rao, D. B.; Nelson, H. G.

    1978-01-01

    As a part of a program for the development of a sulfur probe for monitoring the sulfur potential in coal gasification reactors, an investigation was conducted regarding the efficiency of the solid electrolyte cell Ar+H2+H2S/CaS+CaF2+(Pt)//CaF2//Pt)+CaF2+CaS/H2S+H2+Ar. A demonstration is provided of the theory, design, and operation of a solid-state sulfur probe based on CaF2 electrolyte. It was found that the cell responds to changes in sulfur potential in a manner predicted by the Nernst equation. The response time of the cell at 1225 K, after a small change in temperature or gas composition, was 2.5 Hr, while at a lower temperature of 990 K the response time was approximately 9 hr. The cell emf was insensitive to a moderate increase in the flow rate of the test gas and/or the reference gas. The exact factors affecting the slow response time of galvanic cells based on a CaF2 electrolyte have not yet been determined. The rate-limiting steps may be either the kinetics of electrode reactions or the rate of transport through the electrolyte.

  14. Monitoring of bentonite pore water with a probe based on solid-state microsensors.

    PubMed

    Orozco, Jahir; Baldi, Antoni; Martín, Pedro L; Bratov, Andrei; Jiménez, Cecilia

    2006-10-01

    Repositories for the disposal of radioactive waste generally rely on a multi-barrier system to isolate the waste from the biosphere. This multi-barrier system typically comprises the natural geological barrier provided by the repository host rock and its surroundings and an engineered barrier system (EBS). Bentonite is being studied as an appropriated porous material for an EBS to prevent or delay the release and transport of radionuclides towards biosphere. The study of pore water chemistry within bentonite barriers will permit to understand the transport phenomena of radionuclides and obtain a database of the bentonite-water interaction processes. In this work, the measurement of some chemical parameters in bentonite pore water using solid-state microsensors is proposed. Those sensors are well suited for this application since in situ measurements are feasible and they are robust enough for the long periods of time that monitoring is needed in an EBS. A probe containing an ISFET (ion sensitive field effect transistor) for measuring pH, and platinum microelectrodes for measuring conductivity and redox potential was developed, together with the required instrumentation, to study the chemical changes in a test cell with compacted bentonite. Response features of the sensors' probe and instrumentation performance in synthetic samples with compositions similar to those present in bentonite barriers are reported. Measurements of sensors stability in a test cell are also presented. PMID:17723733

  15. Dynamic nuclear polarization solid-state NMR in heterogeneous catalysis research

    DOE PAGESBeta

    Kobayashi, Takeshi; Perras, Frédéric A.; Slowing, Igor I.; Sadow, Aaron D.; Pruski, Marek

    2015-10-20

    In this study, a revolution in solid-state nuclear magnetic resonance (SSNMR) spectroscopy is taking place, attributable to the rapid development of high-field dynamic nuclear polarization (DNP), a technique yielding sensitivity improvements of 2–3 orders of magnitude. This higher sensitivity in SSNMR has already impacted materials research, and the implications of new methods on catalytic sciences are expected to be profound.

  16. Dynamic nuclear polarization solid-state NMR in heterogeneous catalysis research

    SciTech Connect

    Kobayashi, Takeshi; Perras, Frédéric A.; Slowing, Igor I.; Sadow, Aaron D.; Pruski, Marek

    2015-10-20

    In this study, a revolution in solid-state nuclear magnetic resonance (SSNMR) spectroscopy is taking place, attributable to the rapid development of high-field dynamic nuclear polarization (DNP), a technique yielding sensitivity improvements of 2–3 orders of magnitude. This higher sensitivity in SSNMR has already impacted materials research, and the implications of new methods on catalytic sciences are expected to be profound.

  17. Investigating the interaction between peptides of the amphipathic helix of Hcf106 and the phospholipid bilayer by solid-state NMR spectroscopy.

    PubMed

    Zhang, Lei; Liu, Lishan; Maltsev, Sergey; Lorigan, Gary A; Dabney-Smith, Carole

    2014-01-01

    The chloroplast twin arginine translocation (cpTat) system transports highly folded precursor proteins into the thylakoid lumen using the protonmotive force as its only energy source. Hcf106, as one of the core components of the cpTat system, is part of the precursor receptor complex and functions in the initial precursor-binding step. Hcf106 is predicted to contain a single amino terminal transmembrane domain followed by a Pro-Gly hinge, a predicted amphipathic α-helix (APH), and a loosely structured carboxy terminus. Hcf106 has been shown biochemically to insert spontaneously into thylakoid membranes. To better understand the membrane active capabilities of Hcf106, we used solid-state NMR spectroscopy to investigate those properties of the APH. In this study, synthesized peptides of the predicted Hcf106 APH (amino acids 28-65) were incorporated at increasing mol.% into 1-palmitoyl-2-oleoyl-sn-glycero-phosphocholine (POPC) and POPC/MGDG (monogalactosyldiacylglycerol; mole ratio 85:15) multilamellar vesicles (MLVs) to probe the peptide-lipid interaction. Solid-state (31)P NMR and (2)H NMR spectroscopic experiments revealed that the peptide perturbs the headgroup and the acyl chain regions of phospholipids as indicated by changes in spectral lineshape, chemical shift anisotropy (CSA) line width, and (2)H order SCD parameters. In addition, the comparison between POPC MLVs and POPC/MGDG MLVs indicated that the lipid bilayer composition affected peptide perturbation of the lipids, and such perturbation appeared to be more intense in a system more closely mimicking a thylakoid membrane. PMID:24144541

  18. EASY-GOING DUMBO on-spectrometer optimisation of phase modulated homonuclear decoupling sequences in solid-state NMR

    NASA Astrophysics Data System (ADS)

    Grimminck, Dennis L. A. G.; Vasa, Suresh K.; Meerts, W. Leo; Kentgens, Arno P. M.; Brinkmann, Andreas

    2011-06-01

    A one-step many-parameter optimisation scheme for phase modulated proton homonuclear decoupling sequences in solid-state NMR is presented. Phase modulations, parameterised by DUMBO Fourier coefficients, were optimised using a Covariance Matrix Adaptation Evolution Strategies algorithm. Our method, denoted EASY-GOING DUMBO, starts with featureless spectra and optimises proton-proton decoupling, during either proton or carbon signal detection. Optimisations at moderate sample magic angle spinning (MAS) frequencies and medium radio-frequency (rf) field strengths resulted in solutions closely resembling (e)DUMBO. Application of EASY-GOING DUMBO for optimisation at very high 680 kHz rf field strength, 12.5 kHz MAS on a 400 MHz NMR spectrometer resulted in a new solution, with competitively resolved proton spectra.

  19. Design of a Heterogeneous Catalyst Based on Cellulose Nanocrystals for Cyclopropanation: Synthesis and Solid-State NMR Characterization.

    PubMed

    Liu, Jiquan; Plog, Andreas; Groszewicz, Pedro; Zhao, Li; Xu, Yeping; Breitzke, Hergen; Stark, Annegret; Hoffmann, Rudolf; Gutmann, Torsten; Zhang, Kai; Buntkowsky, Gerd

    2015-08-24

    Heterogeneous dirhodium(II) catalysts based on environmentally benign and biocompatible cellulose nanocrystals (CNC-Rh2) as support material were obtained by ligand exchange between carboxyl groups on the CNC surface and Rh2(OOCCF3)4, as was confirmed by solid-state (19)F and (13)C NMR spectroscopy. On average, two CF3COO(-) groups are replaced during ligand exchange, which is consistent with quantitative analysis by a combination of (19)F NMR spectroscopy and thermogravimetry. CNC-Rh2 catalysts performed well in a model cyclopropanation reaction, in spite of the low dirhodium(II) content on the CNC surface (0.23 mmol g(-1)). The immobilization through covalent bonding combined with the separate locations of binding positions and active sites of CNC-Rh2 guarantees a high stability against leaching and allows the recovery and reuse of the catalyst during the cyclopropanation reaction. PMID:26179865

  20. Studies of interactions between silane coupling agents and cellulose fibers with liquid and solid-state NMR.

    PubMed

    Salon, Marie-Christine B; Gerbaud, Guillaume; Abdelmouleh, M; Bruzzese, Cécile; Boufi, Sami; Belgacem, Mohamed N

    2007-06-01

    The hydrolysis of three alkoxy-silane coupling agents, gamma-methacryloxypropyl trimethoxy silane (MPS), gamma-aminopropyl triethoxy silane (APS), and gamma-diethylenetriaminopropyl trimethoxy silane (TAS), was carried out in ethanol/water solutions (80/20 w/w) at different pH values and followed by 1H, 13C and 29Si NMR spectroscopy. Acidic media were found to stabilize the hydrolyzed forms. As expected, the formation of silanol groups was followed by their self-condensation to generate oligomeric structures, yielding, ultimately, solid homopolycondensated structures, as analyzed by 29Si and 13C high-resolution solid-state NMR. Hydrolyzed MPS in acidic media was then successfully adsorbed onto a cellulose surface and the ensuing substrates submitted to thermal treatment at 110-120 degrees C under reduced pressure, in order to create covalent bonds between cellulose and the coupling agent. PMID:17431857

  1. αB-Crystallin. A Hybrid Solid-State/Solution-State NMR Investigation Reveals Structural Aspects of the Heterogeneous Oligomer

    SciTech Connect

    Jehle, Stefan; van Rossum, Barth; Stout, Joseph R.; Noguchi, Satoshi M.; Falber, Katja; Rehbein, Kristina; Oschkinat, Hartmut; Klevit, Rachel E.; Rajagopal, Ponni

    2008-11-14

    Atomic-level structural information on αB-Crystallin (αB), a prominent member of the small heat-shock protein family, has been a challenge to obtain due its polydisperse oligomeric nature. We show that magic-angle spinning solid-state NMR can be used to obtain high-resolution information on an ~580-kDa human αB assembled from 175-residue 20-kDa subunits. An ~100-residue α-crystallin domain is common to all small heat-shock proteins, and solution-state NMR was performed on two different α- crystallin domain constructs isolated from αB. In vitro, the chaperone-like activities of full-length αB and the isolated α-crystallin domain are identical. Chemical shifts of the backbone and Cβ resonances have been obtained for residues 64–162 (α-crystallin domain plus part of the C-terminus) in αB and the isolated α-crystallin domain by solid-state and solution-state NMR, respectively. Both sets of data strongly predict six β-strands in the α-crystallin domain. A majority of residues in the α-crystallin domain have similar chemical shifts in both solid-state and solution-state, indicating similar structures for the domain in its isolated and oligomeric forms. Sites of intersubunit interaction are identified from chemical shift differences that cluster to specific regions of the α-crystallin domain. Multiple signals are observed for the resonances of M68 in the oligomer, identifying the region containing this residue as existing in heterogeneous environments within αB. Evidence for a novel dimerization motif in the human α-crystallin domain is obtained by a comparison of (i) solid-state and solution-state chemical shift data and (ii) 1H–15N heteronuclear single quantum coherence spectra as a function of pH. The isolated α-crystallin domain undergoes a dimer–monomer transition over the pH range 7.5–6.8. This steep pHdependent switch may be important for αB to function optimally (e.g., to preserve the filament integrity of cardiac muscle proteins such as actin and desmin during cardiac ischemia, which is accompanied by acidosis).

  2. Crystallinity and compositional changes in carbonated apatites: Evidence from {sup 31}P solid-state NMR, Raman, and AFM analysis

    SciTech Connect

    McElderry, John-David P.; Zhu, Peizhi; Mroue, Kamal H.; Xu, Jiadi; Pavan, Barbara; Fang, Ming; Zhao, Guisheng; McNerny, Erin; Kohn, David H.; Franceschi, Renny T.; Holl, Mark M.Banaszak; Tecklenburg, Mary M.J.; Ramamoorthy, Ayyalusamy; Morris, Michael D.

    2013-10-15

    Solid-state (magic-angle spinning) NMR spectroscopy is a useful tool for obtaining structural information on bone organic and mineral components and synthetic model minerals at the atomic-level. Raman and {sup 31}P NMR spectral parameters were investigated in a series of synthetic B-type carbonated apatites (CAps). Inverse {sup 31}P NMR linewidth and inverse Raman PO{sub 4}{sup 3−}ν{sub 1} bandwidth were both correlated with powder XRD c-axis crystallinity over the 0.3–10.3 wt% CO{sub 3}{sup 2−} range investigated. Comparison with bone powder crystallinities showed agreement with values predicted by NMR and Raman calibration curves. Carbonate content was divided into two domains by the {sup 31}P NMR chemical shift frequency and the Raman phosphate ν{sub 1} band position. These parameters remain stable except for an abrupt transition at 6.5 wt% carbonate, a composition which corresponds to an average of one carbonate per unit cell. This near-binary distribution of spectroscopic properties was also found in AFM-measured particle sizes and Ca/P molar ratios by elemental analysis. We propose that this transition differentiates between two charge-balancing ion-loss mechanisms as measured by Ca/P ratios. These results define a criterion for spectroscopic characterization of B-type carbonate substitution in apatitic minerals. - Graphical abstract: Carbonated apatite shows an abrupt change in spectral (NMR, Raman) and morphological (AFM) properties at a composition of about one carbonate substitution per unit cell. Display Omitted - Highlights: • Crystallinity (XRD), particle size (AFM) of carbonated apatites and bone mineral. • Linear relationships among crystallinity, {sup 31}P NMR and Raman inverse bandwidths. • Low and high carbonated apatites use different charge-balancing ion-loss mechanism.

  3. Spatially resolved homonuclear solid-state NMR. III. Magic-echo and rotary-echo phase-encoding imaging

    NASA Astrophysics Data System (ADS)

    Demco, D. E.; Hafner, S.; Kimm1ch, R.

    Two novel magic-echo pulse sequences are reported which are suitable for solid-state NMR imaging using the phase-encoding Fourier transform principle. With the magic- echo phase-encoding solid imaging (MEPSI) technique, ordinary gradients of the external magnetic field are employed. The rotary- echo phase-encoding solid imaging (REPSI) method, on the other hand, is based on gradients of the amplitude of the radiofrequency irradiated to the sample during the magic sandwich pulse in a certain analogy to rotating-frame zeugmatography. The use of magic echoes in solid-state NMR imaging experiments has two crucial advantages. The signal intensity in principle is recovered without incoherence losses. Second, the intervals during which the phase-encoding evolution takes place can be chosen much longer than with ordinary solid-echo experiments. Total encoding times in the order of milliseconds are feasible even with rigid solids. Thus the encoding efficiency of liquid-state imaging is approached. With given gradients, be they main field gradients in the case of MEPSI or gradients of the radiofrequency amplitude in the case of REPSI, therefore, a much better spatial resolution can be achieved than with other phase-encoding techniques. Theoretical treatments and test experiments are presented.

  4. (31)P Solid-State NMR study of the chemical setting process of a dual-paste injectable brushite cements.

    PubMed

    Legrand, A P; Sfihi, H; Lequeux, N; Lemaître, J

    2009-10-01

    The composition and evolution of a brushite-type calcium phosphate cement was investigated by Solid-State NMR and X-ray during the setting process. The cement is obtained by mixing beta-tricalcium phosphate [Ca(3)(PO(4))(2), beta-TCP] and monocalcium phosphate monohydrate [Ca(H(2)PO(4))(2).H(2)O, MCPM] in presence of water, with formation of dicalcium phosphate dihydrate or brushite [CaHPO(2).2H(2)O, DCPD]. Analysis of the initial beta-TCP paste has shown the presence of beta-calcium pyrophosphate [Ca(2)P(2)O(7), beta-CPy] and that of the initial MCPM a mixture of MCPM and dicalcium phosphate [CaHPO(4), DCP]. Follow-up of the chemical composition by (31)P Solid-State NMR enables to show that the chemical setting process appeared to reach an end after 20 min. The constant composition observed at the end of the process was similarly determined. PMID:19365821

  5. Solid-state NMR of a protein in a precipitated complex with a full-length antibody.

    PubMed

    Lamley, Jonathan M; Iuga, Dinu; Öster, Carl; Sass, Hans-Juergen; Rogowski, Marco; Oss, Andres; Past, Jaan; Reinhold, Andres; Grzesiek, Stephan; Samoson, Ago; Lewandowski, Józef R

    2014-12-01

    NMR spectroscopy is a prime technique for characterizing atomic-resolution structures and dynamics of biomolecular complexes but for such systems faces challenges of sensitivity and spectral resolution. We demonstrate that the application of (1)H-detected experiments at magic-angle spinning frequencies of >50 kHz enables the recording, in a matter of minutes to hours, of solid-state NMR spectra suitable for quantitative analysis of protein complexes present in quantities as small as a few nanomoles (tens of micrograms for the observed component). This approach enables direct structure determination and quantitative dynamics measurements in domains of protein complexes with masses of hundreds of kilodaltons. Protein-protein interaction interfaces can be mapped out by comparison of the chemical shifts of proteins within solid-state complexes with those of the same constituent proteins free in solution. We employed this methodology to characterize a >300 kDa complex of GB1 with full-length human immunoglobulin, where we found that sample preparation by simple precipitation yields spectra of exceptional quality, a feature that is likely to be shared with some other precipitating complexes. Finally, we investigated extensions of our methodology to spinning frequencies of up to 100 kHz. PMID:25381931

  6. Complementarity of solvent-free MALDI TOF and solid-state NMR spectroscopy in spectral analysis of polylactides.

    PubMed

    Sroka-Bartnicka, Anna; Ciesielski, Włodzimierz; Libiszowski, Jan; Duda, Andrzej; Sochacki, Marek; Potrzebowski, Marek J

    2010-01-01

    We report systematic studies of solvent-free modification of matrix-assisted laser desorption/ionization time-of-flight (SF MALDI-TOF) mass spectrometry in analysis of synthetic polymers employing solid-state NMR spectroscopy as a supporting technique. In the present work oligomeric (M(n) = 4000 g mol(-1)) poly(L-lactide) (PLLA) was employed as a reference sample. The analyte was embedded into four matrixes commonly used in MALDI-TOF analysis of polymers: 1,8-dihydroxy-9-anthracenone (DT), 2,5-dihydroxybenzoic acid (DHB), 2-(4 hydroxyphenylazo)-benzoic acid (HABA), and trans-3-indoleacrylic acid (IAA). Solid-state NMR measurements clearly showed that the initial crystallinity of PLLA had no influence on quality of SF MALDI-TOF spectra since the crystalline structure of the analyte was not preserved during analyte/matrix grinding. Interestingly, the matrix remained crystalline during the sample's preparation. It was also found that, on the contrary to the dried droplet (DD) method, the SF approach leads to highly resolved mass spectra for a large variety of matrixes. Finally, problems of polymorphism and mechanochemical processes that can occur during the analyte/matrix grinding are briefly discussed. PMID:19957929

  7. Specification and visualization of anisotropic interaction tensors in polypeptides and numerical simulations in biological solid-state NMR.

    PubMed

    Bak, Mads; Schultz, Robert; Vosegaard, Thomas; Nielsen, Niels Chr

    2002-01-01

    Software facilitating numerical simulation of solid-state NMR experiments on polypeptides is presented. The Tcl-controlled SIMMOL program reads in atomic coordinates in the PDB format from which it generates typical or user-defined parameters for the chemical shift, J coupling, quadrupolar coupling, and dipolar coupling tensors. The output is a spin system file for numerical simulations, e.g., using SIMPSON (Bak, Rasmussen, and Nielsen, J. Magn. Reson. 147, 296 (2000)), as well as a 3D visualization of the molecular structure, or selected parts of this, with user-controlled representation of relevant tensors, bonds, atoms, peptide planes, and coordinate systems. The combination of SIMPSON and SIMMOL allows straightforward simulation of the response of advanced solid-state NMR experiments on typical nuclear spin interactions present in polypeptides. Thus, SIMMOL may be considered a "sample changer" to the SIMPSON "computer spectrometer" and proves to be very useful for the design and optimization of pulse sequences for application on uniformly or extensively isotope-labeled peptides where multiple-spin interactions need to be considered. These aspects are demonstrated by optimization and simulation of novel DCP and C7 based 2D N(CO)CA, N(CA)CB, and N(CA)CX MAS correlation experiments for multiple-spin clusters in ubiquitin and by simulation of PISA wheels from PISEMA spectra of uniaxially oriented bacteriorhodopsin and rhodopsin under conditions of finite RF pulses and multiple spin interactions. PMID:11820824

  8. Optimization of an absolute sensitivity in a glassy matrix during DNP-enhanced multidimensional solid-state NMR experiments.

    PubMed

    Takahashi, Hiroki; Fernndez-de-Alba, Carlos; Lee, Daniel; Maurel, Vincent; Gambarelli, Serge; Bardet, Michel; Hediger, Sabine; Barra, Anne-Laure; De Pape, Gal

    2014-02-01

    Thanks to instrumental and theoretical development, notably the access to high-power and high-frequency microwave sources, high-field dynamic nuclear polarization (DNP) on solid-state NMR currently appears as a promising solution to enhance nuclear magnetization in many different types of systems. In magic-angle-spinning DNP experiments, systems of interest are usually dissolved or suspended in glass-forming matrices doped with polarizing agents and measured at low temperature (down to ?100K). In this work, we discuss the influence of sample conditions (radical concentration, sample temperature, etc.) on DNP enhancements and various nuclear relaxation times which affect the absolute sensitivity of DNP spectra, especially in multidimensional experiments. Furthermore, DNP-enhanced solid-state NMR experiments performed at 9.4 T are complemented by high-field CW EPR measurements performed at the same magnetic field. Microwave absorption by the DNP glassy matrix is observed even below the glass transition temperature caused by softening of the glass. Shortening of electron relaxation times due to glass softening and its impact in terms of DNP sensitivity is discussed. PMID:24480716

  9. A comparative spectroscopic investigation of three pseudopolymorphs of testosterone using solid-state i.r. and high-resolution solid-state NMR

    NASA Astrophysics Data System (ADS)

    Fletton, Richard A.; Harris, Robin K.; Kenwright, Alan M.; Lancaster, Robert W.; Packer, Kenneth J.; Sheppard, Norman

    Three pseudopolymorphic forms of testosterone have been examined by i.r. and CP/MAS NMR spectroscopies. The transmittance i.r. data clearly distinguish the forms. The NMR work also provides clear distinctions and shows crystallographic splittings for the α form only, in accordance with the X-ray data. The NMR spectra are fully assigned. The effect of crystallisation procedure on the NMR spectra was explored. NMR can be used to quantitatively assess mixtures of the β and δ forms.

  10. Identification of Li-Ion Battery SEI Compounds through (7)Li and (13)C Solid-State MAS NMR Spectroscopy and MALDI-TOF Mass Spectrometry.

    PubMed

    Huff, Laura A; Tavassol, Hadi; Esbenshade, Jennifer L; Xing, Wenting; Chiang, Yet-Ming; Gewirth, Andrew A

    2016-01-13

    Solid-state (7)Li and (13)C MAS NMR spectra of cycled graphitic Li-ion anodes demonstrate SEI compound formation upon lithiation that is followed by changes in the SEI upon delithiation. Solid-state (13)C DPMAS NMR shows changes in peaks associated with organic solvent compounds (ethylene carbonate and dimethyl carbonate, EC/DMC) upon electrochemical cycling due to the formation of and subsequent changes in the SEI compounds. Solid-state (13)C NMR spin-lattice (T1) relaxation time measurements of lithiated Li-ion anodes and reference poly(ethylene oxide) (PEO) powders, along with MALDI-TOF mass spectrometry results, indicate that large-molecular-weight polymers are formed in the SEI layers of the discharged anodes. MALDI-TOF MS and NMR spectroscopy results additionally indicate that delithiated anodes exhibit a larger number of SEI products than is found in lithiated anodes. PMID:26653886

  11. The effects of anticalcification treatments and hydration on the molecular dynamics of bovine pericardium collagen as revealed by 13C solid-state NMR.

    PubMed

    deAzevedo, E R; Ayrosa, A M I B; Faria, G C; Cervantes, H J; Huster, D; Bonagamba, T J; Pitombo, R N M; Rabbani, S R

    2010-09-01

    This article describes a solid-state NMR (SSNMR) investigation of the influence of hydration and chemical cross-linking on the molecular dynamics of the constituents of the bovine pericardium (BP) tissues and its relation to the mechanical properties of the tissue. Samples of natural phenethylamine-diepoxide (DE)- and glutaraldehyde (GL)-fixed BP were investigated by (13)C cross-polarization SSNMR to probe the dynamics of the collagen, and the results were correlated to the mechanical properties of the tissues, probed by dynamical mechanical analysis. For samples of natural BP, the NMR results show that the higher the hydration level the more pronounced the molecular dynamics of the collagen backbone and sidechains, decreasing the tissue's elastic modulus. In contrast, in DE- and GL-treated samples, the collagen molecules are more rigid, and the hydration seems to be less effective in increasing the collagen molecular dynamics and reducing the mechanical strength of the samples. This is mostly attributed to the presence of cross-links between the collagen plates, which renders the collagen mobility less dependent on the water absorption in chemically treated samples. PMID:20641133

  12. Irregular structure of the HIV fusion peptide in membranes demonstrated by solid-state NMR and MD simulations.

    PubMed

    Grasnick, Dorit; Sternberg, Ulrich; Strandberg, Erik; Wadhwani, Parvesh; Ulrich, Anne S

    2011-04-01

    To better understand peptide-induced membrane fusion at a molecular level, we set out to determine the structure of the fusogenic peptide FP23 from the HIV-1 protein gp41 when bound to a lipid bilayer. An established solid-state (19)F nuclear magnetic resonance (NMR) approach was used to collect local orientational constraints from a series of CF(3)-phenylglycine-labeled peptide analogues in macroscopically aligned membranes. Fusion assays showed that these (19)F-labels did not significantly affect peptide function. The NMR spectra were characteristic of well-behaved samples, without any signs of heterogeneity or peptide aggregation at 1:300 in 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC). We can conclude from these NMR data that FP23 has a well-defined (time-averaged) conformation and undergoes lateral diffusion in the bilayer plane, presumably as a monomer or small oligomer. Attempts to evaluate its conformation in terms of various secondary structures, however, showed that FP23 does not form any type of regular helix or β-strand. Therefore, all-atom molecular dynamics (MD) simulations were carried out using the orientational NMR constraints as pseudo-forces to drive the peptide into a stable alignment and structure. The resulting picture suggests that FP23 can adopt multiple β-turns and insert obliquely into the membrane. Such irregular conformation explains why the structure of the fusion peptide could not be reliably determined by any biophysical method so far. PMID:21274707

  13. Photochemically Induced Dynamic Nuclear Polarization Observed by Solid-State NMR in a Uniformly (13)C-Isotope-Labeled Photosynthetic Reaction Center.

    PubMed

    Paul, Shubhajit; Bode, Bela E; Matysik, Jörg; Alia, A

    2015-10-29

    A sample of solubilized and quinone-depleted reaction centers from the purple bacterium Rhodobacter (R.) sphaeroides wild type has been prepared entirely (13)C and (15)N isotope labeled at all positions of the protein as well as of the cofactors. In this sample, the occurrence of the solid-state photo-CIDNP (photochemically induced dynamic nuclear polarization) effect has been probed by (13)C solid-state magic-angle spinning NMR under illumination. Under continuous illumination, signal intensities are modified by the three-spin mixing (TSM) mechanism. Time-resolved illumination experiments reveal the occurrence of light-induced nuclear polarization on the time scale of hundreds of microseconds, initially dominated by the transient polarization of the singlet branch of the radical-pair mechanism. A first kinetic analysis shows that the lifetime of the polarization from the singlet branch, indicated by the enhanced absorptive intensities of the signals from aliphatic carbons, is significantly extended. Upon arrival of the polarization from the triplet decay branch, emissive polarization caused by the TSM mechanism is observed. Also, this arrival is significantly delayed. The decay of TSM polarization occurs in two steps, assigned to intra- and intermolecular spin diffusion. PMID:26110356

  14. Solid-state NMR analysis of coals and shales from the Mesaverde Group, Green River Basin, Wyoming

    SciTech Connect

    Miknis, F.P.; MacGowan, D.B.

    1993-08-01

    Samples of coals and shales from the Almond Formation of the Mesaverde Group, Greater Green River Basin, Wyoming were analyzed using solid-state {sup 13}C nuclear magnetic resonance (NMR) techniques of cross polarization with magic-angle spinning (CP/MAS). The samples were taken from a present-day depth of burial ranging from {approximately}3,000 to {approximately}15,000 ft. In addition, CP/MAS {sup 13}C NMR measurements were made on residues from the hydrous pyrolysis of Almond coal. The hydrous pyrolysis experiments were conducted isothermally for 72 hr in the temperature range of 290 to 360{degree}C (554 to 680{degree}F). In general, the maturation trends observed by NMR for the naturally and artificially matured samples were in agreement with results obtained from other geochemical analyses. The NMR spectra of the naturally matured shale samples showed only a small aliphatic component at depths greater than about 12,000 ft, indicating little capacity for hydrocarbon generation at depths greater than this. Vitrinite reflectance measurements placed the oil window at between 4,500 and 14,500 ft. NMR measurements of the hydrous pyrolysis residues showed a clear loss of aliphatic carbon, relative to the aromatic carbon, with temperature. For the residue obtained from the highest study temperature (360{degree}C/680{degree}F), there was a 60% depletion of the hydrocarbon-producing aliphatic components. The trends in loss of aliphatic carbon with temperature suggested a means of defining a geochemical transformation ratio in terms of the loss of the aliphatic carbon fraction. A good correlation was found between the NMR transformation ratio and the production index determined by Rock-Eval pyrolysis measurements.

  15. Solid-state Hadamard NMR spectroscopy: Simultaneous measurements of multiple selective homonuclear scalar couplings

    NASA Astrophysics Data System (ADS)

    Kakita, Veera Mohana Rao; Kupče, Eriks; Bharatam, Jagadeesh

    2015-02-01

    Unambiguous measurement of homonuclear scalar couplings (J) in multi-spin scalar network systems is not straightforward. Further, the direct measurement of J-couplings is obscured in solid-state samples due to the dipolar and chemical shift anisotropy (CSA)-dominated line broadening, even under the magic angle spinning (MAS). We present a new multiple frequency selective spin-echo method based on Hadamard matrix encoding, for simultaneous measurement of multiple homonuclear scalar couplings (J) in the solid-state. In contrast to the Hadamard encoded selective excitation schemes known for the solution-state, herein the selectivity is achieved during refocusing period. The Hadamard encoded refocusing scheme concurrently allows to create the spin-spin commutation property between number of spin-pairs of choice in uniformly labelled molecules, which, therefore avoids (1) the repetition of the double selective refocusing experiments for each spin-pair and (2) the synthesis of expensive selective labelled molecules. The experimental scheme is exemplified for determining 1JCC and 3JCC values in 13C6L-Histidine.HCl molecule, which are found to be in excellent agreement with those measured in conventional double frequency selective refocusing mode as well as in the solution-state. This method can be simply extended to 2D/3D pulse schemes and be applied to small bio-molecular solids.

  16. Probing Solid-State Nanopores with Light for the Detection of Unlabeled Analytes

    PubMed Central

    2015-01-01

    Nanopore sensing has enabled label-free single-molecule measurements on a wide variety of analytes, including DNA, RNA, and protein complexes. Much progress has been made toward biotechnological applications; however, electrically probing the ion current introduces nonideal noise components. Here we further develop a method to couple an ionic current to a photon-by-photon counting of fluorescent signal from Ca2+-sensitive dyes and demonstrate label-free optical detection of biopolymer translocation through solid-state nanopores using TIRF and confocal microscopy. We show that by fine adjustment of the CaCl2 gradient, EGTA concentration, and voltage, the optical signals can be localized to the immediate vicinity of the pore. Consequently, the noise spectral density distribution in the optical signal exhibits a nearly flat distribution throughout the entire frequency range. With the use of high-speed photon counting devices in confocal microscopy and higher photon count rates using stronger light sources, we can improve the signal-to-noise ratio of signal acquisition, while the use of wide-field imaging in TIRF can allow for simultaneous quantitative imaging of large arrays of nanopores. PMID:25363680

  17. A spectrometer designed for 6.7 and 14.1 T DNP-enhanced solid-state MAS NMR using quasi-optical microwave transmission

    NASA Astrophysics Data System (ADS)

    Pike, Kevin J.; Kemp, Thomas F.; Takahashi, Hiroki; Day, Robert; Howes, Andrew P.; Kryukov, Eugeny V.; MacDonald, James F.; Collis, Alana E. C.; Bolton, David R.; Wylde, Richard J.; Orwick, Marcella; Kosuga, Kosuke; Clark, Andrew J.; Idehara, Toshitaka; Watts, Anthony; Smith, Graham M.; Newton, Mark E.; Dupree, Ray; Smith, Mark E.

    2012-02-01

    A Dynamic Nuclear Polarisation (DNP) enhanced solid-state Magic Angle Spinning (MAS) NMR spectrometer operating at 6.7 T is described and demonstrated. The 187 GHz TE13 fundamental mode of the FU CW VII gyrotron is used as the microwave source for this magnetic field strength and 284 MHz 1H DNP-NMR. The spectrometer is designed for use with microwave frequencies up to 395 GHz (the TE16 second-harmonic mode of the gyrotron) for DNP at 14.1 T (600 MHz 1H NMR). The pulsed microwave output from the gyrotron is converted to a quasi-optical Gaussian beam using a Vlasov antenna and transmitted to the NMR probe via an optical bench, with beam splitters for monitoring and adjusting the microwave power, a ferrite rotator to isolate the gyrotron from the reflected power and a Martin-Puplett interferometer for adjusting the polarisation. The Gaussian beam is reflected by curved mirrors inside the DNP-MAS-NMR probe to be incident at the sample along the MAS rotation axis. The beam is focussed to a ˜1 mm waist at the top of the rotor and then gradually diverges to give much more efficient coupling throughout the sample than designs using direct waveguide irradiation. The probe can be used in triple channel HXY mode for 600 MHz 1H and double channel HX mode for 284 MHz 1H, with MAS sample temperatures ⩾85 K. Initial data at 6.7 T and ˜1 W pulsed microwave power are presented with 13C enhancements of 60 for a frozen urea solution (1H-13C CP), 16 for bacteriorhodopsin in purple membrane (1H-13C CP) and 22 for 15N in a frozen glycine solution (1H-15N CP) being obtained. In comparison with designs which irradiate perpendicular to the rotation axis the approach used here provides a highly efficient use of the incident microwave beam and an NMR-optimised coil design.

  18. A spectrometer designed for 6.7 and 14.1 T DNP-enhanced solid-state MAS NMR using quasi-optical microwave transmission.

    PubMed

    Pike, Kevin J; Kemp, Thomas F; Takahashi, Hiroki; Day, Robert; Howes, Andrew P; Kryukov, Eugeny V; MacDonald, James F; Collis, Alana E C; Bolton, David R; Wylde, Richard J; Orwick, Marcella; Kosuga, Kosuke; Clark, Andrew J; Idehara, Toshitaka; Watts, Anthony; Smith, Graham M; Newton, Mark E; Dupree, Ray; Smith, Mark E

    2012-02-01

    A Dynamic Nuclear Polarisation (DNP) enhanced solid-state Magic Angle Spinning (MAS) NMR spectrometer operating at 6.7 T is described and demonstrated. The 187 GHz TE(13) fundamental mode of the FU CW VII gyrotron is used as the microwave source for this magnetic field strength and 284 MHz (1)H DNP-NMR. The spectrometer is designed for use with microwave frequencies up to 395 GHz (the TE(16) second-harmonic mode of the gyrotron) for DNP at 14.1T (600 MHz (1)H NMR). The pulsed microwave output from the gyrotron is converted to a quasi-optical Gaussian beam using a Vlasov antenna and transmitted to the NMR probe via an optical bench, with beam splitters for monitoring and adjusting the microwave power, a ferrite rotator to isolate the gyrotron from the reflected power and a Martin-Puplett interferometer for adjusting the polarisation. The Gaussian beam is reflected by curved mirrors inside the DNP-MAS-NMR probe to be incident at the sample along the MAS rotation axis. The beam is focussed to a ~1 mm waist at the top of the rotor and then gradually diverges to give much more efficient coupling throughout the sample than designs using direct waveguide irradiation. The probe can be used in triple channel HXY mode for 600 MHz (1)H and double channel HX mode for 284 MHz (1)H, with MAS sample temperatures ≥85 K. Initial data at 6.7 T and ~1 W pulsed microwave power are presented with (13)C enhancements of 60 for a frozen urea solution ((1)H-(13)C CP), 16 for bacteriorhodopsin in purple membrane ((1)H-(13)C CP) and 22 for (15)N in a frozen glycine solution ((1)H-(15)N CP) being obtained. In comparison with designs which irradiate perpendicular to the rotation axis the approach used here provides a highly efficient use of the incident microwave beam and an NMR-optimised coil design. PMID:22218011

  19. Investigations of the structure and "interfacial" surface chemistry of Bioglass (RTM) materials by solid-state multinuclear NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Sarkar, Gautam

    Bioactive materials such as BioglassRTM 45S5 (45% SiO 2, 24.5% CaO, 24.5% Na2O, and 6% P2O5 by weight) are sodium-phosphosilicate glasses containing independent three-dimensional silicate and phosphate networks and Na+ and Ca2+ ions as modifying cations. Due to their bioactivity, these materials are currently used as implants and for other surgical and clinical applications. The bioactivity of BioglassesRTM is due to their unique capability to form chemical bonds to tissues through an octacalciumphosphate (OCP)- and/or hydroxyapatite-like (HA) "interfacial" matrix. The formation of OCP and/or HA is preceded by the formation of a silica-rich surface layer and the subsequent growth of an amorphous calcium phosphate (a-CP) layer. Structural characterization of a series of commercial and synthesized Bioglass materials 45S5 52S, 55S, 60S, and synthesized 17O-labelled "Bioglass materials 45S, 52S, 55S and 60S" have been obtained using solid-state single-pulse magic-angle spinning (SP/MAS) 17O, 23Na, 29Si and 31P NMR. The 17O NMR isotropic chemical shifts and estimates of the quadrupole coupling constants (Cq) [at fixed asymmetry parameter ( hQ ) values of zero] have been obtained from solid-state spin-echo 17O SP/MAS NMR spectra of 17O-labelled "Bioglasses". The simulation results of these spectra reveal the presence of both bridging-oxygens (BO, i.e. ≡ Si-17OSi ≡ ) and non-bridging oxygens (NBO, i.e. ≡ Si-17O-Na+/Ca2+ ) in the silicate networks in these materials. 17O NMR spectra of these Bioglass materials do not show any direct evidence for the presence of BO and NBO atoms in the phosphate units; however, they are expected to be present in small amounts. In vitro reactions of BioglassRTM 45S5, 60S and 77S powders have been used to study the "interfacial" surface chemistry of these materials in simulated body-fluid (SBF, Kyoto or K9 solution) and/or 17O-enriched tris-buffer solution. 29Si and 31P SP/MAS NMR have been used to identify and quantify the extent of formation of surface silica species and follow the formation of phosphate species, respectively, while cross-polarization magic-angle spinning (CP/MAS) 29Si and 31P NMR have provided information about low intensity NMR peaks due to various silicon- and phosphorus-species present in the vicinity of associated protons on the surface of in vitro reacted BioglassRTM materials. The solid-state NMR investigations of the "interfacial" surface reactions of BioglassRTM materials are discussed in the context of the structure of these materials and the influence of this structure on the kinetics and the mechanism of their "interfacial" surface chemistry. (Abstract shortened by UMI.) BioglassRTM, trademark, University of Florida, Gainesville, FL, 32611.

  20. Characterization of zeolite structure and fluorocarbon reactivity using solid state NMR and x-ray powder diffraction

    NASA Astrophysics Data System (ADS)

    Ciraolo, Michael Frank

    The research presented in this thesis involves a combination of techniques used to study the structure and interactions zeolites adsorbed with fluorocarbons. This research is specifically aimed at understanding the processes of adsorption, binding, and reactivity of fluorocarbons on cation exchanged faujasite type zeolites. The solid state ion exchange process has also been studied since it is one way to obtain materials with higher exchange levels, which has been shown to effect adsorption and catalytic activity. To improve the understanding of the adsorption and separation processes a time resolved in-situ synchrotron x-ray powder diffraction study has been undertaken. Since faujasite type zeolites have been found to be effective in separating mixtures of HFC-134 (CF2HCF2H) and HFC-134a (CFH2CF3) isomers, the adsorption of these fluorocarbons on NaY have been studied. It has been shown that both the extent of loading and the kinetics of the sorption process in molecular sieves can be followed using this technique. A model for the binding of hydrochlorofluorocarbon (HCFC) 124a (CF 2HCF2Cl) adsorbed on NaX at 100K has been determined using a combination x-ray and neutron powder diffraction and solid state NMR. Using Rietveld refinement of the diffraction data, the HCFC molecule was found localized in the zeolite cavities bound on either end by sodium cations in the SII and SIII' positions. The model is consistent with hydrogen bonding between the proton of the HCFC and the framework oxygen. The NMR results further confirm the model and are consistent with Na-F binding and HCFC-framework interactions. Solid-state MAS NMR, synchrotron X-ray powder diffraction and a mass spectrometer and gas chromatograph catalysis system have been used to study the reactivity of HCFC-124a (CF2HCF2Cl) on NaX, Zn 2+-exchanged NaX (ZnX) and Rb+-exchanged NaX (RbX). We have chosen to study HCFC-124a (CF2HCF2Cl) since HCFC-124a can undergo both dehydrofluorination and dehydrochlorination reactions. The preference between these two reactions should be related to the relative importance of the interactions with the basic oxygen atoms and the cations. RbX is considered a basic zeolite while ZnX should have more acidic character, which should be reflected in differences in product distribution. The positions of the exchanged cations and the residual sodium cations of both exchanged zeolites have been located and the products formed during reaction have been identified. To investigate possible methods to achieve high levels of cation substitution, NH4Y has been ion exchanged with solid Rb2CO3 to form RbY. This process has been followed with x-ray powder diffraction, mass spectrometry and NMR. It has been shown that exchange begins on grinding and no rubidium oxide or new carbonate phases were observed to form during the exchange. The positions of the cations of the final exchanged RbY product have been located with Rietveld analysis. It has also been shown that 1H and 87Rb MAS NMR is very sensitive to the level of hydration of the zeolite and has been used to follow the solid state exchange process.

  1. Solid-state variable-temperature NMR study of the phase separation of polybutadiene polyurethane zwitterionomers

    NASA Astrophysics Data System (ADS)

    Yang, G.; Chen, Q.; Wang, Y.; Yang, C.; Wu, X.

    1994-07-01

    Polybutadiene polyurethane (PBDPU) zwitterionomers based on 4,4'-diphenylmethane diisocyanate (MDI), methyl-diethanolamine (MDEA), and hydroxy terminated polybutadiene are studied with variable-temperature (VT) wide-line 1H NMR. Spin—spin relaxation times ( T2) and spin—lattice relaxation times ( T1) are measured. It is found that phase separation of PBDPU does not change significantly upon ionization. The initial incorporation of ionization groups destroys the crystallinity of the hard segment while further ionization enhances physical crosslinks in the hard phase. The results are compared with a previous VT NMR study on polyether polyurethane zwitterionomers based on MDI, MDEA and 1000 Da molecular weight polytetramethylene oxide.

  2. Rapid and accurate determination of the lignin content of lignocellulosic biomass by solid-state NMR

    PubMed Central

    Fu, Li; McCallum, Scott A.; Miao, Jianjun; Hart, Courtney; Tudryn, Gregory J.; Zhang, Fuming; Linhardt, Robert J.

    2014-01-01

    Biofuels and biomaterials, produced from lignocellulosic feedstock, require facile access to cellulose and hemicellulose to be competitive with petroleum processing and sugar-based fermentation. Physical-chemical barriers resulting from lignin complicates the hydrolysis biomass into fermentable sugars. Thus, the amount of lignin within a substrate is critical in determining biomass processing. The application of 13C cross-polarization, magic-angle spinning, and solid-state nuclear magnetic resonance for the direct quantification of lignin content in biomass is examined. Using a standard curve constructed from pristine lignin and cellulose, the lignin content of a biomass sample is accurately determined through direct measurement without chemical or enzymatic pre-treatment. PMID:25404762

  3. A multinuclear solid state NMR spectroscopic study of the structural evolution of disordered calcium silicate sol-gel biomaterials.

    PubMed

    Lin, Zhongjie; Jones, Julian R; Hanna, John V; Smith, Mark E

    2015-01-28

    Disordered sol-gel prepared calcium silicate biomaterials show significant, composition dependent ability to bond with bone. Bone bonding is attributed to rapid hydroxycarbonate apatite (HCA) formation on the glass surface after immersion in body fluid (or implantation). Atomic scale details of the development of the structure of (CaO)x(SiO2)1-x (x = 0.2, 0.3 and 0.5) under heat treatment and subsequent dissolution in simulated body fluid (SBF) are revealed through a multinuclear solid state NMR approach using one-dimensional (17)O, (29)Si, (31)P and (1)H. Central to this study is the combination of conventional static and magic angle spinning (MAS) and two-dimensional (2D) triple quantum (3Q) (17)O NMR experiments that can readily distinguish and quantify the bridging (BOs) and non-bridging (NBOs) oxygens in the silicate network. Although soluble calcium is present in the sol, the (17)O NMR results reveal that the sol-gel produced network structure is initially dominated by BOs after gelation, aging and drying (e.g. at 120 °C), indicating a nanoscale mixture of the calcium salt and a predominantly silicate network. Only once the calcium salt is decomposed at elevated temperatures do the Ca(2+) ions become available to break BO. Apatite forming ability in SBF depends strongly on the surface OH and calcium content. The presence of calcium aids HCA formation via promotion of surface hydration and the ready availability of Ca(2+) ions. (17)O NMR shows the rapid loss of NBOs charge balanced by calcium as it is leached into the SBF. The formation of nanocrystalline, partially ordered HCA can be detected via(31)P NMR. This data indicates the importance of achieving the right balance of BO/NBO for optimal biochemical response and network properties. PMID:25494341

  4. Acceleration of natural-abundance solid-state MAS NMR measurements on bone by paramagnetic relaxation from gadolinium-DTPA

    NASA Astrophysics Data System (ADS)

    Mroue, Kamal H.; Zhang, Rongchun; Zhu, Peizhi; McNerny, Erin; Kohn, David H.; Morris, Michael D.; Ramamoorthy, Ayyalusamy

    2014-07-01

    Reducing the data collection time without affecting the signal intensity and spectral resolution is one of the major challenges for the widespread application of multidimensional nuclear magnetic resonance (NMR) spectroscopy, especially in experiments conducted on complex heterogeneous biological systems such as bone. In most of these experiments, the NMR data collection time is ultimately governed by the proton spin-lattice relaxation times (T1). For over two decades, gadolinium(III)-DTPA (Gd-DTPA, DTPA = Diethylene triamine pentaacetic acid) has been one of the most widely used contrast-enhancement agents in magnetic resonance imaging (MRI). In this study, we demonstrate that Gd-DTPA can also be effectively used to enhance the longitudinal relaxation rates of protons in solid-state NMR experiments conducted on bone without significant line-broadening and chemical-shift-perturbation side effects. Using bovine cortical bone samples incubated in different concentrations of Gd-DTPA complex, the 1H T1 values were calculated from data collected by 1H spin-inversion recovery method detected in natural-abundance 13C cross-polarization magic angle spinning (CPMAS) NMR experiments. Our results reveal that the 1H T1 values can be successfully reduced by a factor of 3.5 using as low as 10 mM Gd-DTPA without reducing the spectral resolution and thus enabling faster data acquisition of the 13C CPMAS spectra. These results obtained from 13C-detected CPMAS experiments were further confirmed using 1H-detected ultrafast MAS experiments on Gd-DTPA doped bone samples. This approach considerably improves the signal-to-noise ratio per unit time of NMR experiments applied to bone samples by reducing the experimental time required to acquire the same number of scans.

  5. Solid-state NMR analysis of soil organic matter fractions from integrated physical-chemical extraction

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fractions of soil organic matter (SOM) are usually extracted from soil by either physical (size, density) or chemical (e.g., base, acid) procedures. In this study we used 13C nuclear magnetic resonance (NMR) spectroscopy to chemically characterize the fractions that were obtained by an integrated pr...

  6. Hydration properties of regioselectively etherified celluloses monitored by 2H and 13C solid-state MAS NMR spectroscopy.

    PubMed

    Larsen, Flemming H; Schöbitz, Michael; Schaller, Jens

    2012-06-20

    The hydration properties of 2,3-O-hydroxypropylcellulose (HPC) and 2,3-O-hydroxyethylcellulose (HEC) were analyzed by multi-nuclear solid-state MAS NMR spectroscopy. By 13C single-pulse (SP) MAS and cross-polarization (CP) MAS NMR, differences between the immobile regions and all parts of the polysaccharides were detected as a function of hydration. Complementary information about the water environments was observed by 2H MAS NMR. By this approach it was demonstrated that side chains in 2,3-O-HPC and 2,3-O-HEC were easier to hydrate than the cellulose backbone. Furthermore the motion of water was more restricted (slower) in 2,3-O-HPC than in 2,3-O-HEC. For both polysaccharides the hydration could be explained by a two-step process: in step one increased ordering of the immobile regions occurs after which the entire polymer is hydrated in step two. PMID:24750769

  7. Molecular composition of recycled organic wastes, as determined by solid-state {sup 13}C NMR and elemental analyses

    SciTech Connect

    Eldridge, S.M.; Chen, C.R.; Xu, Z.H.; Nelson, P.N.; Boyd, S.E.; Meszaros, I.; Chan, K.Y.

    2013-11-15

    Highlights: • Model estimated the molecular C components well for most RO wastes. • Molecular nature of organic matter in RO wastes varied widely. • Molecular composition by NMR modelling preferable to extraction techniques. • Some model shortcomings in estimating molecular composition of biochars. • Waste molecular composition important for carbon/nutrient outcomes in soil. - Abstract: Using solid state {sup 13}C NMR data and elemental composition in a molecular mixing model, we estimated the molecular components of the organic matter in 16 recycled organic (RO) wastes representative of the major materials generated in the Sydney basin area. Close correspondence was found between the measured NMR signal intensities and those predicted by the model for all RO wastes except for poultry manure char. Molecular nature of the organic matter differed widely between the RO wastes. As a proportion of organic C, carbohydrate C ranged from 0.07 to 0.63, protein C from <0.01 to 0.66, lignin C from <0.01 to 0.31, aliphatic C from 0.09 to 0.73, carbonyl C from 0.02 to 0.23, and char C from 0 to 0.45. This method is considered preferable to techniques involving imprecise extraction methods for RO wastes. Molecular composition data has great potential as a predictor of RO waste soil carbon and nutrient outcomes.

  8. Synthesis and evaluation of nitroxide-based oligoradicals for low-temperature dynamic nuclear polarization in solid state NMR

    PubMed Central

    Yau, Wai-Ming; Thurber, Kent R.; Tycko, Robert

    2014-01-01

    We describe the synthesis of new nitroxide-based biradical, triradical, and tetraradical compounds and the evaluation of their performance as paramagnetic dopants in dynamic nuclear polarization (DNP) experiments in solid state nuclear magnetic resonance (NMR) spectroscopy with magic-angle spinning (MAS). Under our experimental conditions, which include temperatures in the 25–30 K range, a 9.4 T magnetic field, MAS frequencies of 6.2–6.8 kHz, and microwave irradiation at 264.0 GHz from a 800 mW extended interaction oscillator source, the most effective compounds are triradicals that are related to the previously-described compound DOTOPA-TEMPO (see Thurber et al., 2010), but have improved solubility in glycerol/water solvent near neutral pH. Using these compounds at 30 mM total nitroxide concentration, we observe DNP enhancement factors of 92–128 for cross-polarized 13C NMR signals from 15N,13C-labeled melittin in partially protonated glycerol/water, and build-up times of 2.6–3.8 s for 1H spin polarizations. Net sensitivity enhancements with biradical and tetraradical dopants, taking into account absolute 13C NMR signal amplitudes and build-up times, are approximately 2–4 times lower than with the best triradicals. PMID:24887201

  9. Solid state NMR, MRI and Sir Peter Mansfield: (1) from broad lines to narrow and back again; and (2) a highly tenuous link to landmine detection.

    PubMed

    Garroway, A N

    1999-12-01

    The contributions of Sir Peter Mansfield to MRI are rooted in solid state NMR. I summarize some of the important contributions of Sir Peter to that field, provide a glimpse of the state of the art in multiple-pulse line-narrowing in the early 1970s, and indicate how the earliest MRI efforts at Nottingham flowed from solid state NMR. These line-narrowing methods, providing control over the Hamiltonian governing the dynamics of nuclear spins, continue to evolve and to find new uses. I indicate how some methods and ideas from solid state NMR of the 1970s are at present applied to the detection of explosives in landmines by nuclear quadrupole resonance (NQR). PMID:10628681

  10. Reassessment of MxiH subunit orientation and fold within native Shigella T3SS needles using surface labelling and solid-state NMR

    PubMed Central

    Verasdonck, Joeri; Shen, Da-Kang; Treadgold, Alexander; Arthur, Christopher; Böckmann, Anja; Meier, Beat H.; Blocker, Ariel J.

    2015-01-01

    T3SSs are essential virulence determinants of many Gram-negative bacteria, used to inject bacterial effectors of virulence into eukaryotic host cells. Their major extracellular portion, a ∼50 nm hollow, needle-like structure, is essential to host cell sensing and the conduit for effector secretion. It is formed of a small, conserved subunit arranged as a helical polymer. The structure of the subunit has been studied by electron cryomicroscopy within native polymers and by solid-state NMR in recombinant polymers, yielding two incompatible atomic models. To resolve this controversy, we re-examined the native polymer used for electron cryomicroscopy via surface labelling and solid-state NMR. Our data show the orientation and overall fold of the subunit within this polymer is as established by solid-state NMR for recombinant polymers. PMID:26439285

  11. Reassessment of MxiH subunit orientation and fold within native Shigella T3SS needles using surface labelling and solid-state NMR.

    PubMed

    Verasdonck, Joeri; Shen, Da-Kang; Treadgold, Alexander; Arthur, Christopher; Böckmann, Anja; Meier, Beat H; Blocker, Ariel J

    2015-12-01

    T3SSs are essential virulence determinants of many Gram-negative bacteria, used to inject bacterial effectors of virulence into eukaryotic host cells. Their major extracellular portion, a ∼50 nm hollow, needle-like structure, is essential to host cell sensing and the conduit for effector secretion. It is formed of a small, conserved subunit arranged as a helical polymer. The structure of the subunit has been studied by electron cryomicroscopy within native polymers and by solid-state NMR in recombinant polymers, yielding two incompatible atomic models. To resolve this controversy, we re-examined the native polymer used for electron cryomicroscopy via surface labelling and solid-state NMR. Our data show the orientation and overall fold of the subunit within this polymer is as established by solid-state NMR for recombinant polymers. PMID:26439285

  12. Solid-state NMR analysis of the {beta}-strand orientation of the protofibrils of amyloid {beta}-protein

    SciTech Connect

    Doi, Takashi; Masuda, Yuichi; Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai 980-8578 ; Irie, Kazuhiro; Akagi, Ken-ichi; Monobe, Youko; Imazawa, Takayoshi; Takegoshi, K.

    2012-11-30

    Highlights: Black-Right-Pointing-Pointer The supramolecular structure of A{beta}42 protofibrils was analyzed by solid-state NMR. Black-Right-Pointing-Pointer The Ala-21 residue in the A{beta}42 protofibrils is included in a slightly disordered {beta}-strand. Black-Right-Pointing-Pointer The A{beta}42 protofibrils do not form intermolecular in-register parallel {beta}-sheets. -- Abstract: Alzheimer's disease (AD) is caused by abnormal deposition (fibrillation) of a 42-residue amyloid {beta}-protein (A{beta}42) in the brain. During the process of fibrillation, the A{beta}42 takes the form of protofibrils with strong neurotoxicity, and is thus believed to play a crucial role in the pathogenesis of AD. To elucidate the supramolecular structure of the A{beta}42 protofibrils, the intermolecular proximity of the Ala-21 residues in the A{beta}42 protofibrils was analyzed by {sup 13}C-{sup 13}C rotational resonance experiments in the solid state. Unlike the A{beta}42 fibrils, an intermolecular {sup 13}C-{sup 13}C correlation was not found in the A{beta}42 protofibrils. This result suggests that the {beta}-strands of the A{beta}42 protofibrils are not in an in-register parallel orientation. A{beta}42 monomers would assemble to form protofibrils with the {beta}-strand conformation, then transform into fibrils by forming intermolecular parallel {beta}-sheets.

  13. Hydration properties and phosphorous speciation in native, gelatinized and enzymatically modified potato starch analyzed by solid-state MAS NMR.

    PubMed

    Larsen, Flemming H; Kasprzak, Mirosław M; Lærke, Helle N; Knudsen, Knud Erik B; Pedersen, Sven; Jørgensen, Anne S; Blennow, Andreas

    2013-09-12

    Hydration of granular, gelatinized and molecularly modified states of potato starch in terms of molecular mobility were analyzed by (13)C and (31)P solid-state MAS NMR. Gelatinization (GEL) tremendously reduced the immobile fraction compared to native (NA) starch granules. This effect was enhanced by enzyme-assisted catalytic branching with branching enzyme (BE) or combined BE and β-amylase (BB) catalyzed exo-hydrolysis. Carbons of the glycosidic α-1,6 linkages required high hydration rates before adopting uniform chemical shifts indicating solid-state disorder and poor water accessibility. Comparative analysis of wheat and waxy maize starches demonstrated that starches were similar upon gelatinization independent of botanical origin and that the torsion angles of the glycosidic linkages were averages of the crystalline A and B type structures. In starch suspension phosphorous in immobile regions was only observed in NA starch. Moreover phosphorous was observed in a minor pH-insensitive form and as major phosphate in hydrated GEL and BE starches. PMID:23911477

  14. Easy-Going On-Spectrometer Optimisation of Phase Modulated Homonuclear Decoupling Sequences in Solid-State NMR

    NASA Astrophysics Data System (ADS)

    Grimminck, Dennis L. A. G.; Vasa, Suresh K.; Meerts, W. Leo; Kentgens, P. M.

    2011-06-01

    A global optimisation scheme for phase modulated proton homonuclear decoupling sequences in solid-state NMR is presented. Phase modulations, parameterised by DUMBO Fourier coefficients, were optimized using a Covariance Matrix Adaptation Evolution Strategies algorithm. Our method, denoted EASY-GOING homonuclear decoupling, starts with featureless spectra and optimises proton-proton decoupling, during either proton or carbon signal detection. On the one hand, our solutions closely resemble (e)DUMBO for moderate sample spinning frequencies and medium radio-frequency (rf) field strengths. On the other hand, the EASY-GOING approach resulted in a superior solution, achieving significantly better resolved proton spectra at very high 680 kHz rf field strength. N. Hansen, and A. Ostermeier. Evol. Comput. 9 (2001) 159-195 B. Elena, G. de Paepe, L. Emsley. Chem. Phys. Lett. 398 (2004) 532-538

  15. An efficient method and device for transfer of semisolid materials into solid-state NMR spectroscopy rotors

    NASA Astrophysics Data System (ADS)

    Hisao, Grant S.; Harland, Michael A.; Brown, Robert A.; Berthold, Deborah A.; Wilson, Thomas E.; Rienstra, Chad M.

    2016-04-01

    The study of mass-limited biological samples by magic angle spinning (MAS) solid-state NMR spectroscopy critically relies upon the high-yield transfer of material from a biological preparation into the MAS rotor. This issue is particularly important for maintaining biological activity and hydration of semi-solid samples such as membrane proteins in lipid bilayers, pharmaceutical formulations, microcrystalline proteins and protein fibrils. Here we present protocols and designs for rotor-packing devices specifically suited for packing hydrated samples into Pencil-style 1.6 mm, 3.2 mm standard, and 3.2 mm limited speed MAS rotors. The devices are modular and therefore readily adaptable to other rotor and/or ultracentrifugation tube geometries.

  16. An efficient method and device for transfer of semisolid materials into solid-state NMR spectroscopy rotors.

    PubMed

    Hisao, Grant S; Harland, Michael A; Brown, Robert A; Berthold, Deborah A; Wilson, Thomas E; Rienstra, Chad M

    2016-04-01

    The study of mass-limited biological samples by magic angle spinning (MAS) solid-state NMR spectroscopy critically relies upon the high-yield transfer of material from a biological preparation into the MAS rotor. This issue is particularly important for maintaining biological activity and hydration of semi-solid samples such as membrane proteins in lipid bilayers, pharmaceutical formulations, microcrystalline proteins and protein fibrils. Here we present protocols and designs for rotor-packing devices specifically suited for packing hydrated samples into Pencil-style 1.6mm, 3.2mm standard, and 3.2mm limited speed MAS rotors. The devices are modular and therefore readily adaptable to other rotor and/or ultracentrifugation tube geometries. PMID:26905816

  17. 2D 31P solid state NMR spectroscopy, electronic structure and thermochemistry of PbP7

    NASA Astrophysics Data System (ADS)

    Benndorf, Christopher; Hohmann, Andrea; Schmidt, Peer; Eckert, Hellmut; Johrendt, Dirk; Schäfer, Konrad; Pöttgen, Rainer

    2016-03-01

    Phase pure polycrystalline PbP7 was prepared from the elements via a lead flux. Crystalline pieces with edge-lengths up to 1 mm were obtained. The assignment of the previously published 31P solid state NMR spectrum to the seven distinct crystallographic sites was accomplished by radio-frequency driven dipolar recoupling (RFDR) experiments. As commonly found in other solid polyphosphides there is no obvious correlation between the 31P chemical shift and structural parameters. PbP7 decomposes incongruently under release of phosphorus forming liquid lead as remainder. The thermal decomposition starts at T>550 K with a vapor pressure almost similar to that of red phosphorus. Electronic structure calculations reveal PbP7 as a semiconductor according to the Zintl description and clearly shows the stereo-active Pb-6s2 lone pairs in the electron localization function ELF.

  18. Solid-state NMR spectroscopy reveals that water is nonessential to the core structure of alpha-synuclein fibrils.

    PubMed

    Kloepper, Kathryn D; Hartman, Kevin L; Ladror, Daniel T; Rienstra, Chad M

    2007-11-29

    Protein aggregation is implicated in the etiology of numerous neurodegenerative diseases. An understanding of aggregation mechanisms is enhanced by atomic-resolution structural information, of which relatively little is currently available. Lewy bodies, the pathological hallmark of Parkinson's disease, contain large quantities of fibrillar alpha-synuclein (AS). Here we present solid-state NMR spectroscopy studies of dried AS fibrils. The spectra have high resolution and sensitivity, and the site-resolved chemical shifts agree very well with those previously observed for hydrated fibrils. The conserved chemical shifts indicate that bulk water is nonessential to the fibril core structure. Moreover, the sample preparation procedure yields major improvements in spectral sensitivity, without compromising spectral resolution. This advance will greatly assist the atomic-resolution structural analysis of AS fibrils. PMID:17985869

  19. Dipolar Assisted Assignment Protocol (DAAP) for MAS solid-state NMR of Rotationally Aligned Membrane Proteins in Phospholipid Bilayers

    PubMed Central

    Das, Bibhuti B.; Zhang, Hua; Opella, Stanley J.

    2014-01-01

    A method for making resonance assignments in magic angle spinning solid-state NMR spectra of membrane proteins that utilizes the range of hetero-nuclear dipolar coupling frequencies in combination with conventional chemical shift based assignment methods is demonstrated. The dipolar assisted assignment protocol (DAAP) takes advantage of the rotational alignment of the membrane proteins in liquid crystalline phospholipid bilayers. Improved resolution is obtained by combining the magnetically inequivalent heteronuclear dipolar frequencies with isotropic chemical shift frequencies. Spectra with both dipolar and chemical shift frequency axes assist with resonance assignments. DAAP can be readily extended to three- and four- dimensional experiments and to include both backbone and side chain sites in proteins. PMID:24698983

  20. The structure, dynamics and orientation of antimicrobial peptides in membranes by multidimensional solid-state NMR spectroscopy.

    PubMed

    Bechinger, B

    1999-12-15

    Linear peptide antibiotics have been isolated from amphibians, insects and humans and used as templates to design cheaper and more potent analogues for medical applications. Peptides such as cecropins or magainins are < or = 40 amino acids in length. Many of them have been prepared by solid-phase peptide synthesis with isotopic labels incorporated at selected sites. Structural analysis by solid-state NMR spectroscopy and other biophysical techniques indicates that these peptide antibiotics strongly interact with lipid membranes. In bilayer environments they exhibit amphipathic alpha-helical conformations and alignments of the helix axis parallel to the membrane surface. This contrasts the transmembrane orientations observed for alamethicin or gramicidin A. Models that have been proposed to explain the antibiotic and pore-forming activities of membrane-associated peptides, as well as other experimental results, include transmembrane helical bundles, wormholes, carpets, detergent-like effects or the in-plane diffusion of peptide-induced bilayer instabilities. PMID:10590307

  1. Solid-State NMR Studies of Fossil Fuels using One- and Two-Dimensional Methods at High Magnetic Field

    SciTech Connect

    Althaus, Stacey M.; Mao, Kanmi; Kennedy, Gordon J.; Pruski, Marek

    2012-06-24

    We examine the opportunities offered by advancements in solid-state NMR (SSNMR) methods, which increasingly rely on the use of high magnetic fields and fast magic angle spinning (MAS), in the studies of coals and other carbonaceous materials. The sensitivity of one- and two-dimensional experiments tested on several Argonne Premium coal samples is only slightly lower than that of traditional experiments performed at low magnetic fields in large MAS rotors, since higher receptivity per spin and the use of 1H detection of low-gamma nuclei can make up for most of the signal loss due to the small rotor size. The advantages of modern SSNMR methodology in these studies include improved resolution, simplicity of pulse sequences, and the possibility of using J-coupling during mixing.

  2. Tripeptides on Gold Nanoparticles: Structural Differences between Two Reverse Sequences as Determined by Solid-State NMR and DFT Calculations.

    PubMed

    Karki, Ichhuk; Wang, Hong; Geise, Natalie R; Wilson, Brendan W; Lewis, James P; Gullion, Terry

    2015-09-10

    The reverse-sequence peptides CysAlaAla and AlaAlaCys may attach to gold nanoparticles through the thiol group, and they differ primarily by whether the charged amino or the carboxylate group is proximal to the sulfur. Alanine residues in these peptides are not expected to interact significantly with the gold surface and serve to place a large separation between the amino and carboxylate groups. Solid-state NMR experiments and DFT calculations were performed to explore the structural differences between CysAlaAla on gold nanoparticles and AlaAlaCys on gold nanoparticles. It is found that the relative positions between the thiol, amino, and carboxylate groups strongly influences the structures of the peptide-gold systems. CysAlaAla orients parallel to the gold surface in a monolayer fashion, whereas AlaAlaCys forms an interdigitating bilayer-like structure that is oriented upright relative to the gold surface. PMID:26308986

  3. High-resolution structural insights into bone: a solid-state NMR relaxation study utilizing paramagnetic doping.

    PubMed

    Mroue, Kamal H; MacKinnon, Neil; Xu, Jiadi; Zhu, Peizhi; McNerny, Erin; Kohn, David H; Morris, Michael D; Ramamoorthy, Ayyalusamy

    2012-09-27

    The hierarchical heterogeneous architecture of bone imposes significant challenges to structural and dynamic studies conducted by traditional biophysical techniques. High-resolution solid-state nuclear magnetic resonance (SSNMR) spectroscopy is capable of providing detailed atomic-level structural insights into such traditionally challenging materials. However, the relatively long data-collection time necessary to achieve a reliable signal-to-noise ratio (S/N) remains a major limitation for the widespread application of SSNMR on bone and related biomaterials. In this study, we attempt to overcome this limitation by employing the paramagnetic relaxation properties of copper(II) ions to shorten the (1)H intrinsic spin-lattice (T(1)) relaxation times measured in natural-abundance (13)C cross-polarization (CP) magic-angle-spinning (MAS) NMR experiments on bone tissues for the purpose of accelerating the data acquisition time in SSNMR. To this end, high-resolution solid-state (13)C CPMAS experiments were conducted on type I collagen (bovine tendon), bovine cortical bone, and demineralized bovine cortical bone, each in powdered form, to measure the (1)H T(1) values in the absence and in the presence of 30 mM Cu(II)(NH(4))(2)EDTA. Our results show that the (1)H T(1) values were successfully reduced by a factor of 2.2, 2.9, and 3.2 for bovine cortical bone, type I collagen, and demineralized bone, respectively, without reducing the spectral resolution and thus enabling faster data acquisition. In addition, paramagnetic quenching of particular (13)C NMR resonances on exposure to Cu(2+) ions in the absence of mineral was also observed, potentially suggesting the relative proximity of three main amino acids in the protein backbone (glycine, proline, and alanine) to the bone mineral surface. PMID:22953757

  4. Characterizing the secondary protein structure of black widow dragline silk using solid-state NMR and X-ray diffraction.

    PubMed

    Jenkins, Janelle E; Sampath, Sujatha; Butler, Emily; Kim, Jihyun; Henning, Robert W; Holland, Gregory P; Yarger, Jeffery L

    2013-10-14

    This study provides a detailed secondary structural characterization of major ampullate dragline silk from Latrodectus hesperus (black widow) spiders. X-ray diffraction results show that the structure of black widow major ampullate silk fibers is comprised of stacked β-sheet nanocrystallites oriented parallel to the fiber axis and an amorphous region with oriented (anisotropic) and isotropic components. The combination of two-dimensional (2D) (13)C-(13)C through-space and through-bond solid-state NMR experiments provide chemical shifts that are used to determine detailed information about the amino acid motif secondary structure in black widow spider dragline silk. Individual amino acids are incorporated into different repetitive motifs that make up the majority of this protein-based biopolymer. From the solid-state NMR measurements, we assign distinct secondary conformations to each repetitive amino acid motif and, hence, to the amino acids that make up the motifs. Specifically, alanine is incorporated in β-sheet (poly(Alan) and poly(Gly-Ala)), 3(1)-helix (poly(Gly-Gly-Xaa), and α-helix (poly(Gln-Gln-Ala-Tyr)) components. Glycine is determined to be in β-sheet (poly(Gly-Ala)) and 3(1)-helical (poly(Gly-Gly-X(aa))) regions, while serine is present in β-sheet (poly(Gly-Ala-Ser)), 3(1)-helix (poly(Gly-Gly-Ser)), and β-turn (poly(Gly-Pro-Ser)) structures. These various motif-specific secondary structural elements are quantitatively correlated to the primary amino acid sequence of major ampullate spidroin 1 and 2 (MaSp1 and MaSp2) and are shown to form a self-consistent model for black widow dragline silk. PMID:24024617

  5. Effect of Ca:Mg ratio on precipitated P species identified using 31P solid state NMR

    NASA Astrophysics Data System (ADS)

    Manimel Wadu, M.

    2009-04-01

    M.C.W. Manimel Wadu1, O.O Akinremi1, S. Kroeker2 1Department of Soil Science, University of Manitoba, Winnipeg, R3T 2N2, Canada 2Department of Chemistry, University of Manitoba, Winnipeg, R3T 2N2, Canada Agronomic efficiency of added P fertilizer is reduced by the precipitation reactions with the exchangeable Ca and Mg in calcareous soils. We hypothesized that the ratio of Ca to Mg on the soil exchange complex will affect the species of P that is precipitated and its solubility in the soil. A laboratory experiment was conducted using a model calcareous soil system which was composed of resin (Amberlite IRP69) and sand coated with CaCO3 packed into a column. The resin was pre saturated with Ca and Mg in order to achieve five different saturation ratios of Ca:Mg approximately as 100:0, 70:30, 50:50, 30:70 and 0:100. Monoammonium Phosphate was applied to the soil surface to simulate one-dimensional diffusive transport. The column was then incubated for 2 weeks. Chemical analysis for water and acid soluble P, pH, NH4, Ca and Mg was performed on 2mm sections of the soil to a depth of 10 cm. This paper will present and discuss the distribution of P along the soil column. Unlike similar studies that have speculated on the precipitation of P, this study will identify and quantify the P species that is formed using 31P solid state NMR technique. Such knowledge will be helpful in understanding the effect of Ca and Mg on P availability in calcareous system and the role of each cation on P precipitation. Key words: P fertilizers, Ca, Mg, model system, solid state NMR

  6. Uncovering the triggers for GPCR activation using solid-state NMR spectroscopy.

    PubMed

    Kimata, Naoki; Reeves, Philip J; Smith, Steven O

    2015-04-01

    G protein-coupled receptors (GPCRs) span cell membranes with seven transmembrane helices and respond to a diverse array of extracellular signals. Crystal structures of GPCRs have provided key insights into the architecture of these receptors and the role of conserved residues. However, the question of how ligand binding induces the conformational changes that are essential for activation remains largely unanswered. Since the extracellular sequences and structures of GPCRs are not conserved between receptor subfamilies, it is likely that the initial molecular triggers for activation vary depending on the specific type of ligand and receptor. In this article, we describe NMR studies on the rhodopsin subfamily of GPCRs and propose a mechanism for how retinal isomerization switches the receptor to the active conformation. These results suggest a general approach for determining the triggers for activation in other GPCR subfamilies using NMR spectroscopy. PMID:25797010

  7. Uncovering the triggers for GPCR activation using solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Kimata, Naoki; Reeves, Philip J.; Smith, Steven O.

    2015-04-01

    G protein-coupled receptors (GPCRs) span cell membranes with seven transmembrane helices and respond to a diverse array of extracellular signals. Crystal structures of GPCRs have provided key insights into the architecture of these receptors and the role of conserved residues. However, the question of how ligand binding induces the conformational changes that are essential for activation remains largely unanswered. Since the extracellular sequences and structures of GPCRs are not conserved between receptor subfamilies, it is likely that the initial molecular triggers for activation vary depending on the specific type of ligand and receptor. In this article, we describe NMR studies on the rhodopsin subfamily of GPCRs and propose a mechanism for how retinal isomerization switches the receptor to the active conformation. These results suggest a general approach for determining the triggers for activation in other GPCR subfamilies using NMR spectroscopy.

  8. Structure of Oxide Glasses under Compression and Confinement: Insights from High-resolution Solid-state NMR and Non-resonant Inelastic X-ray Scattering

    NASA Astrophysics Data System (ADS)

    Lee, S.

    2013-12-01

    The atomic and nano-scale structure of non-crystalline oxides with varying degree of confinement and compression (pressure) are essential to understand the atomic origins of thermo-mechanical properties and kinetic stability of these materials in the earth's surfaces and interiors. Despite this importance, the effect of confinement and pressure on the nature of bonding in the non-crystalline oxides have remained one of the challenging problems in mineral physics and condensed matter physics due to lack of suitable experimental probes. Advances in element-specific experimental probes, such as non-resonant inelastic x-ray scattering (NRIXS) and high resolution solid-state NMR combined with the first principle calculations have revealed structural details of bonding transitions of amorphous oxides under compression and confinement (e.g. Lee, Rev. Min. Geochem. 2013 accepted; Phys. Rev. Letts, 2013 accepted; Proc. Nat. Aca. Sci. 2011, 108, 6847; Kim and Lee, Geochim. Cosmochim Acta. In press; Yi and Lee, Am Min 2012, 97, 897). Here, we present the key recent progress by NRIXS and NMR into the pressure and confinement-induced bonding transitions in non-crystalline oxides. Theoretical calculation of K-edge NRIXS spectrum took into consideration crystallographically-distinct sites and revealed that the edge features systematically shift to higher energy with increasing degree of densification in atomic arrangement in the polymorphs (from enstatite, perovskite, to post-perovskite). On the basis of these results, multi-nuclear (B, O, Li, Ca), multi-edge (K-, L-, and M-) XRS studies of diverse low z-oxide glasses indicated the pressure-induced increases in the fraction of triply coordinated oxygen above 20 GPa and showed evidence for the topologically driven densification in multi-component basaltic glasses. Finally, we report the first high-resolution solid-state NMR results for the amorphous oxides under confinement where the degree of structural disorder tends to decrease with increasing degree of confinement. These results allow us to microscopically confine the geochemical processes involving interactions between melts and fluids.

  9. Solid-State 87Sr NMR Spectroscopy at Natural Abundance and High Magnetic Field Strength.

    PubMed

    Faucher, Alexandra; Terskikh, Victor V; Ye, Eric; Bernard, Guy M; Wasylishen, Roderick E

    2015-12-10

    Twenty-five strontium-containing solids were characterized via (87)Sr NMR spectroscopy at natural abundance and high magnetic field strength (B0 = 21.14 T). Strontium nuclear quadrupole coupling constants in these compounds are sensitive to the strontium site symmetry and range from 0 to 50.5 MHz. An experimental (87)Sr chemical shift scale is proposed, and available data indicate a chemical shift range of approximately 550 ppm, from -200 to +350 ppm relative to Sr(2+)(aq). In general, magnetic shielding increased with strontium coordination number. Experimentally measured chemical shift anisotropy is reported for stationary samples of solid powdered SrCl2·6H2O, SrBr2·6H2O, and SrCO3, with δaniso((87)Sr) values of +28, +26, and -65 ppm, respectively. NMR parameters were calculated using CASTEP, a gauge including projector augmented wave (GIPAW) DFT-based program, which addresses the periodic nature of solids using plane-wave basis sets. Calculated NMR parameters are in good agreement with those measured. PMID:26565918

  10. Solid-state NMR characterization of the mineral searlesite and its detection in complex synthesis mixtures.

    PubMed

    Fyfe, C A; Skibsted, J; Schwieger, W

    2001-11-01

    The mineral searlesite (NaBSi(2)O(5)(OH)(2)) was synthesized and characterized by (1)H, (11)B, (23)Na, and (29)Si magic-angle spinning (MAS) NMR spectroscopy. From these spectra, the (11)B and (23)Na quadrupole coupling parameters and isotropic chemical shifts and the (29)Si chemical shift anisotropies have been precisely determined. These parameters are all consistent with the local environments obtained from the crystal structure for searlesite from X-ray diffraction, and they demonstrate that the synthetic sample has a high degree of both short- and long-range order. Furthermore, these anisotropic parameters are found to provide a unique fingerprinting of searlesite in complex mixtures where the presence of this mineral is not anticipated. This is demonstrated for product mixtures formed in attempts to incorporate boron in the structures of the layer silicates magadiite and kenyaite. These mixtures have been investigated by (11)B, (23)Na, and (29)Si MAS NMR which clearly reveal that the samples are mixtures of searlesite and magadiite/kenyaite and that searlesite production consumes all of the boron in these synthesis mixtures. However, the (29)Si MAS NMR spectra of these mixtures indicate that the presence of boron in the reaction mixtures nevertheless has an important influence on the quality of the magadiite and kenyaite layer silicates produced. PMID:11681903

  11. Trimethylphosphine-Assisted Surface Fingerprinting of Metal Oxide Nanoparticle by (31)P Solid-State NMR: A Zinc Oxide Case Study.

    PubMed

    Peng, Yung-Kang; Ye, Lin; Qu, Jin; Zhang, Li; Fu, Yingyi; Teixeira, Ivo F; McPherson, Ian James; He, Heyong; Tsang, Shik Chi Edman

    2016-02-24

    Nano metal oxides are becoming widely used in industrial, commercial and personal products (semiconductors, optics, solar cells, catalysts, paints, cosmetics, sun-cream lotions, etc.). However, the relationship of surface features (exposed planes, defects and chemical functionalities) with physiochemical properties is not well studied primarily due to lack of a simple technique for their characterization. In this study, solid state (31)P MAS NMR is used to map surfaces on various ZnO samples with the assistance of trimethylphosphine (TMP) as a chemical probe. As similar to XRD giving structural information on a crystal, it is demonstrated that this new surface-fingerprint technique not only provides qualitative (chemical shift) but also quantitative (peak intensity) information on the concentration and distribution of cations and anions, oxygen vacancies and hydroxyl groups on various facets from a single deconvoluted (31)P NMR spectrum. On the basis of this technique, a new mechanism for photocatalytic •OH radical generation from direct surface-OH oxidation is revealed, which has important implications regarding the safety of using nano oxides in personal care products. PMID:26812527

  12. {sup 45}Sc Solid State NMR studies of the silicides ScTSi (T=Co, Ni, Cu, Ru, Rh, Pd, Ir, Pt)

    SciTech Connect

    Harmening, Thomas; Eckert, Hellmut; Fehse, Constanze M.; Sebastian, C. Peter; Poettgen, Rainer

    2011-12-15

    The silicides ScTSi (T=Fe, Co, Ni, Cu, Ru, Rh, Pd, Ir, Pt) were synthesized by arc-melting and characterized by X-ray powder diffraction. The structures of ScCoSi, ScRuSi, ScPdSi, and ScIrSi were refined from single crystal diffractometer data. These silicides crystallize with the TiNiSi type, space group Pnma. No systematic influences of the {sup 45}Sc isotropic magnetic shift and nuclear electric quadrupolar coupling parameters on various structural distortion parameters calculated from the crystal structure data can be detected. {sup 45}Sc MAS-NMR data suggest systematic trends in the local electronic structure probed by the scandium atoms: both the electric field gradients and the isotropic magnetic shifts relative to a 0.2 M aqueous Sc(NO{sub 3}){sub 3} solution decrease with increasing valence electron concentration and within each T group the isotropic magnetic shift decreases monotonically with increasing atomic number. The {sup 45}Sc nuclear electric quadrupolar coupling constants are generally well reproduced by quantum mechanical electric field gradient calculations using the WIEN2k code. Highlights: Black-Right-Pointing-Pointer Arc-melting synthesis of silicides ScTSi. Black-Right-Pointing-Pointer Single crystal X-ray data of ScCoSi, ScRuSi, ScPdSi, and ScIrSi. Black-Right-Pointing-Pointer {sup 45}Sc solid state NMR of silicides ScTSi.

  13. Catalytic roles of βLys87 in tryptophan synthase: (15)N solid state NMR studies.

    PubMed

    Caulkins, Bethany G; Yang, Chen; Hilario, Eduardo; Fan, Li; Dunn, Michael F; Mueller, Leonard J

    2015-09-01

    The proposed mechanism for tryptophan synthase shows βLys87 playing multiple catalytic roles: it bonds to the PLP cofactor, activates C4' for nucleophilic attack via a protonated Schiff base nitrogen, and abstracts and returns protons to PLP-bound substrates (i.e. acid-base catalysis). ε-¹⁵N-lysine TS was prepared to access the protonation state of βLys87 using ¹⁵N solid-state nuclear magnetic resonance (SSNMR) spectroscopy for three quasi-stable intermediates along the reaction pathway. These experiments establish that the protonation state of the ε-amino group switches between protonated and neutral states as the β-site undergoes conversion from one intermediate to the next during catalysis, corresponding to mechanistic steps where this lysine residue has been anticipated to play alternating acid and base catalytic roles that help steer reaction specificity in tryptophan synthase catalysis. This article is part of a Special Issue entitled: Cofactor-dependent proteins: evolution, chemical diversity and bio-applications. Guest Editors: Andrea Mozzarelli and Loredano Pollegioni. PMID:25688830

  14. A unified heteronuclear decoupling strategy for magic-angle-spinning solid-state NMR spectroscopy

    SciTech Connect

    Equbal, Asif; Bjerring, Morten; Nielsen, Niels Chr. E-mail: ncn@inano.au.dk; Madhu, P. K. E-mail: ncn@inano.au.dk

    2015-05-14

    A unified strategy of two-pulse based heteronuclear decoupling for solid-state magic-angle spinning nuclear magnetic resonance is presented. The analysis presented here shows that different decoupling sequences like two-pulse phase-modulation (TPPM), X-inverse-X (XiX), and finite pulse refocused continuous wave (rCW{sup A}) are basically specific solutions of a more generalized decoupling scheme which incorporates the concept of time-modulation along with phase-modulation. A plethora of other good decoupling conditions apart from the standard, TPPM, XiX, and rCW{sup A} decoupling conditions are available from the unified decoupling approach. The importance of combined time- and phase-modulation in order to achieve the best decoupling conditions is delineated. The consequences of different indirect dipolar interactions arising from cross terms comprising of heteronuclear and homonuclear dipolar coupling terms and also those between heteronuclear dipolar coupling and chemical-shift anisotropy terms are presented in order to unfold the effects of anisotropic interactions under different decoupling conditions. Extensive numerical simulation results are corroborated with experiments on standard amino acids.

  15. An experimental and theoretical NMR study of NH-benzimidazoles in solution and in the solid state: proton transfer and tautomerism

    PubMed Central

    Nieto, Carla I; Cabildo, Pilar; García, M Ángeles; Elguero, José

    2014-01-01

    Summary This paper reports the 1H, 13C and 15N NMR experimental study of five benzimidazoles in solution and in the solid state (13C and 15N CPMAS NMR) as well as the theoretically calculated (GIAO/DFT) chemical shifts. We have assigned unambiguously the "tautomeric positions" (C3a/C7a, C4/C7 and C5/C6) of NH-benzimidazoles that, in some solvents and in the solid state, appear different (blocked tautomerism). In the case of 1H-benzimidazole itself we have measured the prototropic rate in HMPA-d 18. PMID:25161719

  16. Advanced solid-state NMR characterization of marine dissolved organic matter isolated using the coupled reverse osmosis/electrodialysis method.

    PubMed

    Mao, Jingdong; Kong, Xueqian; Schmidt-Rohr, Klaus; Pignatello, Joseph J; Perdue, E Michael

    2012-06-01

    Advanced (13)C solid-state techniques were employed to investigate the major structural characteristics of two surface-seawater dissolved organic matter (DOM) samples isolated using the novel coupled reverse osmosis/electrodialysis method. The NMR techniques included quantitative (13)C direct polarization/magic angle spinning (DP/MAS) and DP/MAS with recoupled dipolar dephasing, (13)C cross-polarization/total sideband suppression (CP/TOSS), (13)C chemical shift anisotropy filter, CH, CH(2), and CH(n) selection, two-dimensional (1)H-(13)C heteronuclear correlation NMR (2D HETCOR), 2D HETCOR combined with dipolar dephasing, and (15)N cross-polarization/magic angle spinning (CP/MAS). The two samples (Coastal and Marine DOM) were collected at the mouth of the Ogeechee River and in the Gulf Stream, respectively. The NMR results indicated that they were structurally distinct. Coastal DOM contained significantly more aromatic and carbonyl carbons whereas Marine DOM was markedly enriched in alkoxy carbon (e.g., carbohydrate-like moieties). Both samples contained significant amide N, but Coastal DOM had nitrogen bonded to aromatic carbons. Our dipolar-dephased spectra indicated that a large fraction of alkoxy carbons were not protonated. For Coastal DOM, our NMR results were consistent with the presence of the major structural units of (1) carbohydrate-like moieties, (2) lignin residues, (3) peptides or amino sugars, and (4) COO-bonded alkyls. For Marine DOM, they were (1) carbohydrate-like moieties, (2) peptides or amino sugars, and (3) COO-bonded alkyls. In addition, both samples contained significant amounts of nonpolar alkyl groups. The potential sources of the major structural units of DOM were discussed in detail. Nonprotonated O-alkyl carbon content was proposed as a possible index of humification. PMID:22553962

  17. Cation substitution in β-tricalcium phosphate investigated using multi-nuclear, solid-state NMR

    SciTech Connect

    Grigg, Andrew T.; Mee, Martin; Mallinson, Phillip M.; Fong, Shirley K.; Gan, Zhehong; Dupree, Ray; Holland, Diane

    2014-04-01

    The substitution of aluminium, gallium and sodium cations into β-tricalcium phosphate (β-TCP; Ca{sub 3}(PO{sub 4}){sub 2}) has been investigated, and the Ca sites involved successfully determined, using a combination of 1D {sup 31}P, {sup 27}Al, {sup 71}Ga, {sup 23}Na and {sup 43}Ca (natural abundance) NMR and 2D {sup 27}Al({sup 31}P), {sup 71}Ga({sup 31}P) and {sup 23}Na({sup 31}P) rotary-resonance-recoupled heteronuclear multiple-quantum correlation (R{sup 3}-HMQC) NMR. Over the compositional range studied, substitution of Ca{sup 2+} by Al{sup 3+} or Ga{sup 3+} was observed only on the Ca(5) site, whilst substitution by Na{sup +} was confined to the Ca(4) site. Some AlPO{sub 4} or GaPO{sub 4} second phase was observed at the highest doping levels in the Al{sup 3+} and Ga{sup 3+} substituted samples. - Graphical abstract: 2D contour plots with skyline projections showing recoupling of {sup 27}Al, {sup 71}Ga and {sup 23}Na to different {sup 31}P sites. - Highlights: • β-Ca{sub 3}(PO{sub 4}){sub 2} has been prepared pure and also with Al{sup 3+}, Ga{sup 3+} and Na{sup +} substituents. • Multi-nuclear 1D NMR and heteronuclear X({sup 31}P) recoupling have been used. • Models for substitution correctly predict site preference and occupancy. • Progressive changes in {sup 31}P spectra have been explained. • Al{sup 3+} and Ga{sup 3+} substitute onto the Ca(5) site, and Na{sup +} onto the Ca(4) site.

  18. Probing spin density and local structure in the Prussian blue analogues CsCd[Fe/Co(CN)6]·0.5H2O and Cd3[Fe/Co(CN)6]2·15H2O with solid-state MAS NMR spectroscopy.

    PubMed

    Flambard, Alexandrine; Köhler, Frank H; Lescouëzec, Rodrigue; Revel, Bertrand

    2011-10-01

    Magic-angle spinning (MAS) NMR spectroscopy is used to study the local structure and spin delocalisation in Prussian blue analogues (PBAs). We selected two common archetypes of PBAs (A(I)M(II)[M(III)(CN)(6)]·xH(2)O and M(II)(3)[M(III)(CN)(6)](2)·xH(2)O, in which A(I) is an alkali ion, and M(II) and M(III) are transition-metal ions) that exhibit similar cubic frameworks but different microscopic structures. Whereas the first type of PBA contains interstitial alkali ions and does not exhibit any [M(III)(CN)(6)](3-) vacancies, the second type of PBA exhibits [M(III)(CN)(6)](3-) vacancies, but does not contain inserted alkali ions. In this study, we selected Cd(II) as a divalent metal in order to use the (113)Cd nuclei (I=1/2) as a probe of the local structure. Here, we present a complete MAS NMR study on two series of PBAs of the formulas Cd(II)(3)[Fe(III)(x)Co(III)(1-x)(CN)(6)](2)·15H(2)O with x=0 (1), 0.25 (2), 0.5 (3), 0.75 (4) and 1 (5), and CsCd(II)[Fe(III)(x)Co(III)(1-x)(CN)(6)]·0.5H(2)O with x=0 (6), 0.25 (7), 0.5 (8), 0.75 (9) and 1 (10). Interestingly, the presence of Fe(III) magnetic centres in the vicinity of the cadmium sites has a magnifying-glass effect on the NMR spectrum: it induces a striking signal spread such that the resolution is notably improved compared to that achieved for the diamagnetic PBAs. By doping the sample with varying amounts of diamagnetic Co(III) and comparing the NMR spectra of both types of PBAs, we have been able to give a view of the structure which is complementary to that usually obtained from X-ray diffraction studies. In particular, this study has shown that the vacancies are not randomly distributed in the mesoporous PBAs. Moreover the cadmium chemical shift, which is a measure of the hyperfine coupling, allows the estimation of the spin density on the cadmium nucleus, and consequently, the elucidation of the spin delocalisation mechanism in these compounds along with its dependency on structural parameters. PMID:21882267

  19. Preparation of highly and generally enriched mammalian tissues for solid state NMR.

    PubMed

    Wong, Veronica Wai Ching; Reid, David G; Chow, Wing Ying; Rajan, Rakesh; Green, Maggie; Brooks, Roger A; Duer, Melinda J

    2015-10-01

    An appreciable level of isotope labelling is essential for future NMR structure elucidation of mammalian biomaterials, which are either poorly expressed, or unexpressable, using micro-organisms. We present a detailed protocol for high level (13)C enrichment even in slow turnover murine biomaterials (fur keratin), using a customized diet supplemented with commercial labelled algal hydrolysate and formulated as a gel to minimize wastage, which female mice consumed during pregnancy and lactation. This procedure produced approximately eightfold higher fur keratin labelling in pups, exposed in utero and throughout life to label, than in adults exposed for the same period, showing both the effectiveness, and necessity, of this approach. PMID:26407607

  20. Identification of different oxygen species in oxide nanostructures with (17)O solid-state NMR spectroscopy.

    PubMed

    Wang, Meng; Wu, Xin-Ping; Zheng, Sujuan; Zhao, Li; Li, Lei; Shen, Li; Gao, Yuxian; Xue, Nianhua; Guo, Xuefeng; Huang, Weixin; Gan, Zhehong; Blanc, Frédéric; Yu, Zhiwu; Ke, Xiaokang; Ding, Weiping; Gong, Xue-Qing; Grey, Clare P; Peng, Luming

    2015-02-01

    Nanostructured oxides find multiple uses in a diverse range of applications including catalysis, energy storage, and environmental management, their higher surface areas, and, in some cases, electronic properties resulting in different physical properties from their bulk counterparts. Developing structure-property relations for these materials requires a determination of surface and subsurface structure. Although microscopy plays a critical role owing to the fact that the volumes sampled by such techniques may not be representative of the whole sample, complementary characterization methods are urgently required. We develop a simple nuclear magnetic resonance (NMR) strategy to detect the first few layers of a nanomaterial, demonstrating the approach with technologically relevant ceria nanoparticles. We show that the (17)O resonances arising from the first to third surface layer oxygen ions, hydroxyl sites, and oxygen species near vacancies can be distinguished from the oxygen ions in the bulk, with higher-frequency (17)O chemical shifts being observed for the lower coordinated surface sites. H2 (17)O can be used to selectively enrich surface sites, allowing only these particular active sites to be monitored in a chemical process. (17)O NMR spectra of thermally treated nanosized ceria clearly show how different oxygen species interconvert at elevated temperature. Density functional theory calculations confirm the assignments and reveal a strong dependence of chemical shift on the nature of the surface. These results open up new strategies for characterizing nanostructured oxides and their applications. PMID:26601133

  1. Solid state NMR of SiO 2 nanotube coated ammonium tartrate crystal

    NASA Astrophysics Data System (ADS)

    Lim, A. R.; Schueneman, G. T.; Novak, B. M.

    1999-04-01

    Ammonium tartrate crystal and SiO 2 nanotube coated ammonium tartrate crystal were studied by 13C CP/MAS NMR, and the structure of two samples were verified using the 13C NMR spectrum. The spin-lattice relaxation times for the carbons in the rotating frame, T1 ρ, have been measured as a function of temperature. All relaxation times of the carbons in the two materials undergo slow motions, i.e. motions on the slow side of the T1 ρ minimum. From these relaxation times, we determine the activation energy for the ammonium tartrate crystal and SiO 2 nanotube coated ammonium tartrate crystal, respectively. The activation energies for the SiO 2 nanotube coated ammonium tartrate crystal were found to be generally higher than those of ammonium tartrate crystal. We think that the higher activation energy for the hydrocarbon in the SiO 2 nanotube coated ammonium tartrate crystal is because of the bonding between the oxygen in the SiO 2 nanotube and the hydroxyl group of the ammonium tartrate crystal.

  2. Identification of different oxygen species in oxide nanostructures with 17O solid-state NMR spectroscopy

    PubMed Central

    Wang, Meng; Wu, Xin-Ping; Zheng, Sujuan; Zhao, Li; Li, Lei; Shen, Li; Gao, Yuxian; Xue, Nianhua; Guo, Xuefeng; Huang, Weixin; Gan, Zhehong; Blanc, Frédéric; Yu, Zhiwu; Ke, Xiaokang; Ding, Weiping; Gong, Xue-Qing; Grey, Clare P.; Peng, Luming

    2015-01-01

    Nanostructured oxides find multiple uses in a diverse range of applications including catalysis, energy storage, and environmental management, their higher surface areas, and, in some cases, electronic properties resulting in different physical properties from their bulk counterparts. Developing structure-property relations for these materials requires a determination of surface and subsurface structure. Although microscopy plays a critical role owing to the fact that the volumes sampled by such techniques may not be representative of the whole sample, complementary characterization methods are urgently required. We develop a simple nuclear magnetic resonance (NMR) strategy to detect the first few layers of a nanomaterial, demonstrating the approach with technologically relevant ceria nanoparticles. We show that the 17O resonances arising from the first to third surface layer oxygen ions, hydroxyl sites, and oxygen species near vacancies can be distinguished from the oxygen ions in the bulk, with higher-frequency 17O chemical shifts being observed for the lower coordinated surface sites. H217O can be used to selectively enrich surface sites, allowing only these particular active sites to be monitored in a chemical process. 17O NMR spectra of thermally treated nanosized ceria clearly show how different oxygen species interconvert at elevated temperature. Density functional theory calculations confirm the assignments and reveal a strong dependence of chemical shift on the nature of the surface. These results open up new strategies for characterizing nanostructured oxides and their applications. PMID:26601133

  3. A New Tool for NMR Crystallography: Complete (13)C/(15)N Assignment of Organic Molecules at Natural Isotopic Abundance Using DNP-Enhanced Solid-State NMR.

    PubMed

    Märker, Katharina; Pingret, Morgane; Mouesca, Jean-Marie; Gasparutto, Didier; Hediger, Sabine; De Paëpe, Gaël

    2015-11-01

    NMR crystallography of organic molecules at natural isotopic abundance (NA) strongly relies on the comparison of assigned experimental and computed NMR chemical shifts. However, a broad applicability of this approach is often hampered by the still limited (1)H resolution and/or difficulties in assigning (13)C and (15)N resonances without the use of structure-based chemical shift calculations. As shown here, such difficulties can be overcome by (13)C-(13)C and for the first time (15)N-(13)C correlation experiments, recorded with the help of dynamic nuclear polarization. We present the complete de novo (13)C and (15)N resonance assignment at NA of a self-assembled 2'-deoxyguanosine derivative presenting two different molecules in the asymmetric crystallographic unit cell. This de novo assignment method is exclusively based on aforementioned correlation spectra and is an important addition to the NMR crystallography approach, rendering firstly (1)H assignment straightforward, and being secondly a prerequisite for distance measurements with solid-state NMR. PMID:26485326

  4. Molecular composition of recycled organic wastes, as determined by solid-state 13C NMR and elemental analyses.

    PubMed

    Eldridge, S M; Chen, C R; Xu, Z H; Nelson, P N; Boyd, S E; Meszaros, I; Chan, K Y

    2013-11-01

    Using solid state (13)C NMR data and elemental composition in a molecular mixing model, we estimated the molecular components of the organic matter in 16 recycled organic (RO) wastes representative of the major materials generated in the Sydney basin area. Close correspondence was found between the measured NMR signal intensities and those predicted by the model for all RO wastes except for poultry manure char. Molecular nature of the organic matter differed widely between the RO wastes. As a proportion of organic C, carbohydrate C ranged from 0.07 to 0.63, protein C from <0.01 to 0.66, lignin C from <0.01 to 0.31, aliphatic C from 0.09 to 0.73, carbonyl C from 0.02 to 0.23, and char C from 0 to 0.45. This method is considered preferable to techniques involving imprecise extraction methods for RO wastes. Molecular composition data has great potential as a predictor of RO waste soil carbon and nutrient outcomes. PMID:23896223

  5. Quantitative analysis of Bordeaux red wine precipitates by solid-state NMR: Role of tartrates and polyphenols.

    PubMed

    Prakash, Shipra; Iturmendi, Nerea; Grelard, Axelle; Moine, Virginie; Dufourc, Erick

    2016-05-15

    Stability of wines is of great importance in oenology matters. Quantitative estimation of dark red precipitates formed in Merlot and Cabernet Sauvignon wine from Bordeaux region for vintages 2012 and 2013 was performed during the oak barrel ageing process. Precipitates were obtained by placing wine at -4°C or 4°C for 2-6 days and monitored by periodic sampling during a one-year period. Spectroscopic identification of the main families of components present in the precipitate powder was performed with (13)C solid-state CPMAS NMR and 1D and 2D solution NMR of partially water re-solubilized precipitates. The study revealed that the amount of precipitate obtained is dependent on vintage, temperature and grape variety. Major components identified include potassium bitartrate, polyphenols, polysaccharides, organic acids and free amino acids. No evidence was found for the presence of proteins. The influence of main compounds found in the precipitates is discussed in relation to wine stability. PMID:26775965

  6. Solid-state dynamics in the closo-carboranes: a (11)B MAS NMR and molecular dynamics study.

    PubMed

    Ahumada, Hernán; Kurkiewicz, Teresa; Thrippleton, Michael J; Wimperis, Stephen

    2015-03-19

    This work explores the dynamic behavior of the three closo-carborane isomers (formula C2B10H12) using modern solid-state magic angle spinning (MAS) NMR techniques and relates the experimental measurements to theoretical results obtained using molecular dynamics simulations. At high temperatures and at B0 = 9.4 T, the (11)B MAS line widths are narrow (40-90 Hz) for the three isomers. The rotational correlation times (τc) calculated by molecular dynamics are on the picosecond time scale, showing a quasi-isotropic rotation at these temperatures, typical for liquid systems. For all three isomers, the values of the (11)B spin-lattice relaxation times (T1) show discontinuities as the temperature is decreased, confirming the phase changes reported in the literature. At low temperatures, the (11)B MAS spectra of all three isomers exhibit much broader lines. The simulations showed that the molecular reorientation was anisotropic around different symmetry axes for each isomer, and this was supported by the values of the reduced quadrupolar parameter PQ(eff) derived from "dynamic shift" measurements using (11)B MQMAS NMR spectroscopy. The behavior of PQ(eff) as a function of temperature for p-carborane suggests that molecular reorientation is about the C5 symmetry axis of the molecule at low temperatures, and this was supported by the molecular dynamics simulations. PMID:25710751

  7. Solid state NMR chemical shift assignment and conformational analysis of a cellulose binding protein facilitated by optimized glycerol enrichment.

    PubMed

    Ivanir, Hadar; Goldbourt, Amir

    2014-07-01

    Magic-angle spinning solid-state NMR has been applied to study CBM3b-Cbh9A (CBM3b), a cellulose binding module protein belonging to family 3b. It is a 146-residue protein having a unique nine-stranded β-sandwich fold, in which 35% of the structure is in a β-sheet conformation and the remainder of the protein is composed of loops and unstructured regions. Yet, the protein can be crystalized and it forms elongated needles. Close to complete chemical shift assignment of the protein was obtained by combining two- and three-dimensional experiments using a fully labeled sample and a glycerol-labeled sample. The use of an optimized protocol for glycerol-based sparse labeling reduces sample preparation costs and facilitates the assignment of the large number of aromatic signals in this protein. Conformational analysis shows good correlation between the NMR-predicted secondary structure and the reported X-ray crystal structure, in particular in the structured regions. Residues which show high B-factor values are situated mainly in unstructured regions, and are missing in our spectra indicating conformational flexibility rather than heterogeneity. Interestingly, long-range contacts, which could be clearly detected for tyrosine residues, could not be observed for aromatic phenylalanine residues pointing into the hydrophobic core, suggesting possible high ring mobility. These studies will allow us to further investigate the cellulose-bound form of CBM proteins. PMID:24824437

  8. Efficient and stable reconstitution of the ABC transporter BmrA for solid-state NMR studies.

    PubMed

    Kunert, Britta; Gardiennet, Carole; Lacabanne, Denis; Calles-Garcia, Daniel; Falson, Pierre; Jault, Jean-Michel; Meier, Beat H; Penin, François; Böckmann, Anja

    2014-01-01

    We present solid-state NMR sample preparation and first 2D spectra of the Bacillus subtilis ATP-binding cassette (ABC) transporter BmrA, a membrane protein involved in multidrug resistance. The homodimeric 130-kDa protein is a challenge for structural characterization due to its membrane-bound nature, size, inherent flexibility and insolubility. We show that reconstitution of this protein in lipids from Bacillus subtilis at a lipid-protein ratio of 0.5 w/w allows for optimal protein insertion in lipid membranes with respect to two central NMR requirements, high signal-to-noise in the spectra and sample stability over a time period of months. The obtained spectra point to a well-folded protein and a highly homogenous preparation, as witnessed by the narrow resonance lines and the signal dispersion typical for the expected secondary structure distribution of BmrA. This opens the way for studies of the different conformational states of the transporter in the export cycle, as well as on interactions with substrates, via chemical-shift fingerprints and sequential resonance assignments. PMID:25988146

  9. Efficient and stable reconstitution of the ABC transporter BmrA for solid-state NMR studies

    PubMed Central

    Kunert, Britta; Gardiennet, Carole; Lacabanne, Denis; Calles-Garcia, Daniel; Falson, Pierre; Jault, Jean-Michel; Meier, Beat H.; Penin, François; Böckmann, Anja

    2014-01-01

    We present solid-state NMR sample preparation and first 2D spectra of the Bacillus subtilis ATP-binding cassette (ABC) transporter BmrA, a membrane protein involved in multidrug resistance. The homodimeric 130-kDa protein is a challenge for structural characterization due to its membrane-bound nature, size, inherent flexibility and insolubility. We show that reconstitution of this protein in lipids from Bacillus subtilis at a lipid-protein ratio of 0.5 w/w allows for optimal protein insertion in lipid membranes with respect to two central NMR requirements, high signal-to-noise in the spectra and sample stability over a time period of months. The obtained spectra point to a well-folded protein and a highly homogenous preparation, as witnessed by the narrow resonance lines and the signal dispersion typical for the expected secondary structure distribution of BmrA. This opens the way for studies of the different conformational states of the transporter in the export cycle, as well as on interactions with substrates, via chemical-shift fingerprints and sequential resonance assignments. PMID:25988146

  10. Molecular dynamics of a polyaniline/β-cyclodextrin complex investigated by 13C solid-state NMR.

    PubMed

    Hasegawa, Yuichi; Inoue, Yoshio; Deguchi, Kenzo; Ohki, Shinobu; Tansho, Masataka; Shimizu, Tadashi; Yazawa, Koji

    2012-02-16

    The molecular dynamics of a polyaniline/β-cyclodextrin inclusion complex (PANI/β-CD IC) and its relation with optical properties were investigated using high-resolution solid-state (13)C nuclear magnetic resonance (NMR) and optical absorption spectroscopies. UV-vis measurements revealed a π-π* absorption peak of a PANI film that had a 10 nm blue-shift by inclusion of β-CD, indicating that π-conjugation of PANI was shortened in the IC. Temperature dependent analysis of (13)C NMR spectra and spin-lattice relaxation times (T(1C)) revealed that the inclusion induced acceleration of the twisting motion of the PANI chain. Moreover, two twisting motions attributed to different torsional angle modes were observed following Arrhenius plots of T(1C) measurements, and the twisting frequency and angle increased above -25 °C. These results suggest that the β-CD inclusion weakens the intermolecular π-π interaction and enhances the accompanying twisting motion, consequently leading to a blue-shift of UV-vis absorption. PMID:22233191

  11. Solid state NMR and IR characterization of wood polymer structure in relation to tree provenance.

    PubMed

    Santoni, Ilaria; Callone, Emanuela; Sandak, Anna; Sandak, Jakub; Dirè, Sandra

    2015-03-01

    (13)C nuclear magnetic resonance and mid-infrared spectroscopies were used for characterizing changes in the chemical structure of wood polymers (cellulose, hemicellulose and lignin) in relation to the tree growth location. Samples of three provenances in Europe (Finland, Poland and Italy) were selected for studies. The requirement was to use untreated solid wood samples to minimize any manipulation to the nanostructure of native wood. The results confirm that the chemical and physical properties of samples belonging to the same wood species (Picea abies Karst.) differ due to the origin. Both FT-IR and dynamic NMR spectroscopies were able to correctly discriminate samples originating from three different provenances in Europe. Such methods might be very useful for both, research and understanding of wood microstructure and its variability due to the growth conditions. PMID:25498692

  12. Mechanism of Solid-State Thermolysis of Ammonia Boraine: 15N NMR Study Using Fast Magic-Angle Spinning and Dynamic Nuclear Polarization

    SciTech Connect

    Kobayashi, Takeshi; Gupta, Shalabh; Caporini, Marc A; Pecharsky, Vitalij K; Pruski, Marek

    2014-08-28

    The solid-state thermolysis of ammonia borane (NH3BH3, AB) was explored using state-of-the-art 15N solid-state NMR spectroscopy, including 2D indirectly detected 1H{15N} heteronuclear correlation and dynamic nuclear polarization (DNP)-enhanced 15N{1H} cross-polarization experiments as well as 11B NMR. The complementary use of 15N and 11B NMR experiments, supported by density functional theory calculations of the chemical shift tensors, provided insights into the dehydrogenation mechanism of AB—insights that have not been available by 11B NMR alone. Specifically, highly branched polyaminoborane derivatives were shown to form from AB via oligomerization in the “head-to-tail” manner, which then transform directly into hexagonal boron nitride analog through the dehydrocyclization reaction, bypassing the formation of polyiminoborane.

  13. Quadrupolar Metal Nuclides in Bioinorganic Chemistry: Solid-State NMR Studies

    SciTech Connect

    Lipton, Andrew S.; Ellis, Paul D.; Polenova, Tatyana E.

    2009-09-16

    Metal ions play an important role in bioinorganic chemistry, however, following their respective chemistries is often complicated because several relevant metal ions (such as V5+, Cu1+, Zn2+, and Mg2+) are not always amenable to conventional UV/Vis or EPR spectroscopy. Rather, what we know of these metal sites has come from the characterization of the various compounds and proteins via x-ray crystallographic methods or from using surrogate metal probes for conventional spectroscopy

  14. An alternative solution for computer controlled tuning and matching of existing NMR probes

    NASA Astrophysics Data System (ADS)

    Koczor, Bálint; Sedyó, Inez; Rohonczy, János

    2015-10-01

    Tuning and matching of NMR probes is necessary for many fields of NMR application including temperature dependent NMR, thermoporometry and cryoporometry, or when significantly different types of samples are measured in automation using sample changers. Mismatch of the probe is an especially critical issue in the case of high magnetic fields, polar or ionic solvents, or extreme thermal conditions. Careful tuning is particularly important for quantitative NMR measurements. Manual tuning and matching of the NMR probe is not possible in the case of automated or remotely controlled measurements. Spectrometer manufacturers offer modern probes equipped with automatic tuning/matching mechanics, like Bruker ATM™, suitable for these experiments. The disadvantages of probes with built-in ATM™ are the significantly higher price, and the non-detachable and non-portable construction. Computer controlled tuning and matching is highly desirrable in solid state NMR since no industrial solution has been developed yet for MAS NMR probes. We present an alternative solution for computer controlled tuning and matching of existing Bruker probes. Building costs are significantly lower, since only commercially available components and ICs are used.

  15. An alternative solution for computer controlled tuning and matching of existing NMR probes.

    PubMed

    Koczor, Bálint; Sedyó, Inez; Rohonczy, János

    2015-10-01

    Tuning and matching of NMR probes is necessary for many fields of NMR application including temperature dependent NMR, thermoporometry and cryoporometry, or when significantly different types of samples are measured in automation using sample changers. Mismatch of the probe is an especially critical issue in the case of high magnetic fields, polar or ionic solvents, or extreme thermal conditions. Careful tuning is particularly important for quantitative NMR measurements. Manual tuning and matching of the NMR probe is not possible in the case of automated or remotely controlled measurements. Spectrometer manufacturers offer modern probes equipped with automatic tuning/matching mechanics, like Bruker ATM™, suitable for these experiments. The disadvantages of probes with built-in ATM™ are the significantly higher price, and the non-detachable and non-portable construction. Computer controlled tuning and matching is highly desirrable in solid state NMR since no industrial solution has been developed yet for MAS NMR probes. We present an alternative solution for computer controlled tuning and matching of existing Bruker probes. Building costs are significantly lower, since only commercially available components and ICs are used. PMID:26363581

  16. Hydrothermal carbon from biomass: structural differences between hydrothermal and pyrolyzed carbons via 13C solid state NMR.

    PubMed

    Falco, Camillo; Perez Caballero, Fernando; Babonneau, Florence; Gervais, Christel; Laurent, Guillaume; Titirici, Maria-Magdalena; Baccile, Niki

    2011-12-01

    The objective of this paper is to better describe the structure of the hydrothermal carbon (HTC) process and put it in relationship with the more classical pyrolytic carbons. Indeed, despite the low energetic impact and the number of applications described so far for HTC, very little is known about the structure, reaction mechanism, and the way these materials relate to coals. Are HTC and calcination processes equivalent? Are the structures of the processed materials related to each other in any way? Which is the extent of polyaromatic hydrocarbons (PAH) inside HTC? In this work, the effect of hydrothermal treatment and pyrolysis are compared on glucose, a good model carbohydrate; a detailed single-quantum double-quantum (SQ-DQ) solid state (13)C NMR study of the HTC and calcined HTC is used to interpret the spectral region corresponding to the signal of furanic and arene groups. These data are compared to the spectroscopic signatures of calcined glucose, starch, and xylose. A semiquantitative analysis of the (13)C NMR spectra provides an estimation of the furanic-to-arene ratio which varies from 1:1 to 4:1 according to the processing conditions and carbohydrate employed. In addition, we formulate some hypothesis, validated by DFT (density functional theory) modeling associated with (13)C NMR chemical shifts calculations, about the possible furan-rich structural intermediates that occur in the coalification process leading to condensed polyaromatic structures. In combination with a broad parallel study on the HTC processing conditions effect on glucose, cellulose, and raw biomass (Falco, C.; Baccile, N.; Titirici, M.-M. Green Chem., 2011, DOI: 10.1039/C1GC15742F), we propose a broad reaction scheme and in which we show that, through HTC, it is possible to tune the furan-to-arene ratio composing the aromatic core of the produced HTC carbons, which is not possible if calcination is used alone, in the temperature range below 350 °C. PMID:22050004

  17. Dipole tensor-based atomic-resolution structure determination of a nanocrystalline protein by solid-state NMR

    PubMed Central

    Franks, W. Trent; Wylie, Benjamin J.; Schmidt, Heather L. Frericks; Nieuwkoop, Andrew J.; Mayrhofer, Rebecca-Maria; Shah, Gautam J.; Graesser, Daniel T.; Rienstra, Chad M.

    2008-01-01

    Magic-angle spinning (MAS) solid-state NMR (SSNMR) techniques have emerged in recent years for solving complete structures of uniformly labeled proteins lacking macroscopic order. Strategies used thus far have relied primarily on semiquantitative distance restraints, analogous to the nuclear Overhauser effect (NOE) routinely used in solution NMR. Here, we present a complementary approach for using relative orientations of molecular fragments, determined from dipolar line shapes. Whereas SSNMR distance restraints typically have an uncertainty of ≈1 Å, the tensor-based experiments report on relative vector (pseudobond) angles with precision of a few degrees. By using 3D techniques of this type, vector angle (VEAN) restraints were determined for the majority of the 56-residue B1 immunoglobulin binding domain of protein G [protein GB1 (a total of 47 HN-HN, 49 HN-HC, and 12 HA-HB restraints)]. By using distance restraints alone in the structure calculations, the overall backbone root-mean-square deviation (bbRMSD) was 1.01 ± 0.13 Å (1.52 ± 0.12 Å for all heavy atoms), which improved to 0.49 ± 0.05 Å (1.19 ± 0.07 Å) on the addition of empirical chemical shift [torsion angle likelihood obtained from shift and sequence similarity (TALOS)] restraints. VEAN restraints further improved the ensemble to 0.31 ± 0.06 Å bbRMSD (1.06 ± 0.07 Å); relative to the structure with distances alone, most of the improvement remained (bbRMSD 0.64 ± 0.09 Å; 1.29 ± 0.07 Å) when TALOS restraints were removed before refinement. These results represent significant progress toward atomic-resolution protein structure determination by SSNMR, capabilities that can be applied to a large range of membrane proteins and fibrils, which are often not amenable to solution NMR or x-ray crystallography. PMID:18344321

  18. Magic-angle spinning solid-state NMR of a 144 kDa membrane protein complex: E. coli cytochrome bo3 oxidase.

    PubMed

    Frericks, Heather L; Zhou, Donghua H; Yap, Lai Lai; Gennis, Robert B; Rienstra, Chad M

    2006-09-01

    Recent progress in magic-angle spinning (MAS) solid-state NMR (SSNMR) has enabled multidimensional studies of large, macroscopically unoriented membrane proteins with associated lipids, without the requirement of solubility that limits other structural techniques. Here we present initial sample preparation and SSNMR studies of a 144 kDa integral membrane protein, E. coli cytochrome bo(3) oxidase. The optimized protocol for expression and purification yields approximately 5 mg of the enzymatically active, uniformly (13)C,(15)N-enriched membrane protein complex from each liter of growth medium. The preparation retains endogenous lipids and yields spectra of high sensitivity and resolution, consistent with a folded, homogenous protein. Line widths of isolated signals are less than 0.5 ppm, with a large number of individual resonances resolved in the 2D and 3D spectra. The (13)C chemical shifts, assigned by amino acid type, are consistent with the secondary structure previously observed by diffraction methods. Although the structure is predominantly helical, the percentage of non-helical signals varies among residue types; these percentages agree well between the NMR and diffraction data. Samples show minimal evidence of degradation after several weeks of NMR data acquisition. Use of a triple resonance scroll resonator probe further improves sample stability and enables higher power decoupling, higher duty cycles and more advanced 3D experiments to be performed. These initial results in cytochrome bo(3) oxidase demonstrate that multidimensional MAS SSNMR techniques have sufficient sensitivity and resolution to interrogate selected parts of a very large uniformly (13)C,(15)N-labeled membrane protein. PMID:16964530

  19. Hyperpolarized NMR Probes for Biological Assays

    PubMed Central

    Meier, Sebastian; Jensen, Pernille R.; Karlsson, Magnus; Lerche, Mathilde H.

    2014-01-01

    During the last decade, the development of nuclear spin polarization enhanced (hyperpolarized) molecular probes has opened up new opportunities for studying the inner workings of living cells in real time. The hyperpolarized probes are produced ex situ, introduced into biological systems and detected with high sensitivity and contrast against background signals using high resolution NMR spectroscopy. A variety of natural, derivatized and designed hyperpolarized probes has emerged for diverse biological studies including assays of intracellular reaction progression, pathway kinetics, probe uptake and export, pH, redox state, reactive oxygen species, ion concentrations, drug efficacy or oncogenic signaling. These probes are readily used directly under natural conditions in biofluids and are often directly developed and optimized for cellular assays, thus leaving little doubt about their specificity and utility under biologically relevant conditions. Hyperpolarized molecular probes for biological NMR spectroscopy enable the unbiased detection of complex processes by virtue of the high spectral resolution, structural specificity and quantifiability of NMR signals. Here, we provide a survey of strategies used for the selection, design and use of hyperpolarized NMR probes in biological assays, and describe current limitations and developments. PMID:24441771

  20. Hyperpolarized NMR probes for biological assays.

    PubMed

    Meier, Sebastian; Jensen, Pernille R; Karlsson, Magnus; Lerche, Mathilde H

    2014-01-01

    During the last decade, the development of nuclear spin polarization enhanced (hyperpolarized) molecular probes has opened up new opportunities for studying the inner workings of living cells in real time. The hyperpolarized probes are produced ex situ, introduced into biological systems and detected with high sensitivity and contrast against background signals using high resolution NMR spectroscopy. A variety of natural, derivatized and designed hyperpolarized probes has emerged for diverse biological studies including assays of intracellular reaction progression, pathway kinetics, probe uptake and export, pH, redox state, reactive oxygen species, ion concentrations, drug efficacy or oncogenic signaling. These probes are readily used directly under natural conditions in biofluids and are often directly developed and optimized for cellular assays, thus leaving little doubt about their specificity and utility under biologically relevant conditions. Hyperpolarized molecular probes for biological NMR spectroscopy enable the unbiased detection of complex processes by virtue of the high spectral resolution, structural specificity and quantifiability of NMR signals. Here, we provide a survey of strategies used for the selection, design and use of hyperpolarized NMR probes in biological assays, and describe current limitations and developments. PMID:24441771

  1. Exploration of structure and function in biomolecules through solid-state NMR and computational methods

    NASA Astrophysics Data System (ADS)

    Heider, Elizabeth M.

    Solid-State Nuclear Magnetic Resonance (SSNMR) spectroscopy and quantum mechanical calculations are powerful analysis tools. Leveraged independently, each method yields important nuclear and molecular information. Used in concert, SSNMR and computational techniques provide complementary data about the structure of solids. These methods are particularly useful in characterizing the structures of microcrystalline organic compounds and revealing mechanisms of biological activity. Such applications may possess special relevance in analysis of pharmaceutical products; 90% of all pharmaceuticals are marketed as solids and bioactivity is strongly linked with molecular conformation. Accordingly, this dissertation employs both SSNMR and quantum mechanical computation to study three bioactive molecules: citrinin, two forms of Atrasentan (Abt-627), and paclitaxel (Taxol RTM). First, a computational study is utilized to determine the mechanism for unusual antioxidant activity in citrinin. Here, molecular geometries and bond dissociation enthalpies (BDE) of the citrinin O--H groups are calculated from first principles (ab initio). The total molecular Hamiltonian is determined by approximating the individual contributors to energy including electronic energy and contributions from modes of molecular vibration. This study of citrinin clearly identifies specific reaction sites in the active form, establishing the central role of intramolecular hydrogen bonding in this activity. Notably, it is discovered that citrinin itself is not the active species. Instead, a pair of hydrated Michael addition products of citrinin act as radical scavengers via O--H bond dissociation. Next, two separate compounds of the anticancer drug Abt-627 (form I and form II) are examined via SSNMR. The three principal values of the 13C diagonalized chemical shift tensor are acquired through the high resolution 2D experiment, FIREMAT. Isotropic chemical shift assignments are made utilizing both dipolar dephasing experiments and 1H-- 13C heteronuclear correlation (HETCOR) experiments. A comparison of spectral data confirms the presence of two molecules in the asymmetric unit for form II (Z'=2) and regions of conformational variation between the two forms are posited. Structural rigidity is found throughout both forms and extends into the alkyl groups at the amine with similarties between form I and form II in this moiety. Likely regions of motion are found around the bond axes formed by C1--C5 in form I. This motion is also observed in one of the two molecules of form II. Tensor differences between the two forms at the tetrahydro-pyrrole center indicate that conformational variation between form I and form II exists in the dihedral angles formed by the atoms C14--C13--C3--C2, O--C12--C2--C1, C10--C5--C1--N1 and C21--C20--N1--C4. Finally, SSNMR is applied in conjunction with quantum mechanical calculations in the analysis of a novel polymorph of the anticancer drug paclitaxel. The three dimensional structure of paclitaxel is established through a combination of SSNMR tensor (13C & 15N) and 1H--13C HETCOR data. With two molecules in the asymmetric unit (Z'=2), this represents the first conformational characterization with Z'>1 established solely by SSNMR. Semi-empirical models are constructed and fitted to experimental data by adjusting the conformation of the paclitaxel models and selecting those conformers which minimize the difference between predicted and measured tensors. This computational grid search exhausively samples the conformation of paclitaxel, utilizing more than 600 independent models. HETCOR data at thirteen key positions provide shift assignment to the asymmetric unit for each comparison. The two distinct molecules of the asymmetric unit possess nearly identical baccatin III moieties with matching conformations of the C10 acetyl moiety. Additionally, both are found to exhibit an extended conformation of the phenylisoserine sidechain at the C13 position. Notable differences between the two forms are centered around the rotation axes of O--C13, C2'--C1 ', and C3'--C2'.

  2. Chemical structures of swine-manure chars produced under different carbonization conditions investigated by advanced solid-state 13C nuclear magnetic resonance (NMR) spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two types of swine manure chars, hydrothermally-produced hydrochar and slow-pyrolysis pyrochar, and their raw swine manure solid were characterized using advanced 13C solid-state nuclear magnetic resonance (NMR) spectroscopy. Compared with the parent raw swine manure, both hydrochars and pyrochar di...

  3. Differences between Lignin in Unprocessed Wood, Milled Wood, Mutant Wood, and Extracted Lignin Detected by 13C Solid-State NMR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Solid-state 13C nuclear magnetic resonance (NMR) has been applied to an array of intact and isolated wood samples in order to identify potential structural changes induced by tree age, milling, lignin extraction, or naturally occurring mutations. Included in this study were mature loblolly pine mil...

  4. High Field Solid-State NMR Spectroscopy Investigation of (15)N-Labeled Rosette Nanotubes: Hydrogen Bond Network and Channel-Bound Water.

    PubMed

    Fenniri, Hicham; Tikhomirov, Grigory A; Brouwer, Darren H; Bouatra, Souhaila; El Bakkari, Mounir; Yan, Zhimin; Cho, Jae-Young; Yamazaki, Takeshi

    2016-05-18

    (15)N-labeled rosette nanotubes were synthesized and investigated using high-field solid-state NMR spectroscopy, X-ray diffraction, atomic force microscopy, and electron microscopy. The results established the H-bond network involved in the self-assembly of the nanostructure as well as bound water molecules in the nanotube's channel. PMID:27141817

  5. High-resolution solid-state {sup 13}C and {sup 15}N NMR spectroscopy of pyrazole and 3,5-dimethylpyrazole adsorbed on alumina and silica

    SciTech Connect

    Aguilar-Parrilla, F.; Limbach, H.H.; Claramunt, R.M.

    1994-09-01

    Using pyrazole and 3,5-dimethylpyrazole mixtures with alumina and silica, high-resolution solid state {sup 13}C and {sup 15}N CPMAS NMR was performed to compare the spectra. The NH-N proton tautomers resulting depend strongly on the environment. 70 refs., 8 figs., 4 tabs.

  6. Cholesterol orientation and dynamics in dimyristoylphosphatidylcholine bilayers: a solid state deuterium NMR analysis.

    PubMed Central

    Marsan, M P; Muller, I; Ramos, C; Rodriguez, F; Dufourc, E J; Czaplicki, J; Milon, A

    1999-01-01

    Proton decoupled deuterium NMR spectra of oriented bilayers made of DMPC and 30 mol % deuterated cholesterol acquired at 76.8 MHz (30 degreesC) have provided a set of very accurate quadrupolar splitting for eight C-D bonds of cholesterol. Due to the new precision of the experimental data, the original analysis by. Biochemistry. 23:6062-6071) had to be reconsidered. We performed a systematic study of the influence on the precision and uniqueness of the data-fitting procedure of: (i) the coordinates derived from x-ray, neutron scattering, or force field-minimized structures, (ii) internal mobility, (iii) the axial symmetry hypothesis, and (iv) the knowledge of some quadrupolar splitting assignments. Good agreement between experiment and theory could be obtained only with the neutron scattering structure, for which both axial symmetry hypothesis and full order parameter matrix analysis gave satisfactory results. Finally, this work revealed an average orientation of cholesterol slightly different from those previously published and, most importantly, a molecular order parameter equal to 0.95 +/- 0.01, instead of 0.79 +/- 0.03 previously found for the same system at 30 degreesC. Temperature dependence in the 20-50 degreesC range shows a constant average orientation and a monotonous decrease of cholesterol Smol, with a slope of -0.0016 K-1. A molecular order parameter of 0.89 +/- 0.01 at 30 degreesC was determined for a DMPC/16 mol % of cholesterol. PMID:9876147

  7. Solid state 1H NMR studies of cell wall materials of potatoes

    NASA Astrophysics Data System (ADS)

    Tang, Huiru; Belton, Peter S.; Ng, Annie; Waldron, Keith W.; Ryden, Peter

    1999-04-01

    Cell wall materials from potatoes ( Solanum tuberosum) prepared by two different methods have been studied using NMR proton relaxation times. Spin lattice relaxation in both the rotating and laboratory frames as well as transverse relaxation have been measured over a range of temperatures and hydration levels. It was observed that the sample prepared using a DMSO extraction showed anomalous behaviour of spin lattice relaxation in the laboratory frame probably due to residual solvent in the sample. Spin lattice relaxation in the laboratory frame is the result of hydroxymethyl rotation and another unidentified high frequency motion. In the rotating frame relaxation is adequately explained by hydroxymethyl rotation alone. In neither experiment is methyl group rotation observed, calculation suggests that this is due to the low density of methyl groups in the sample. Non-freezing water in potato cell walls, α-cellulose and pectin was found about 0.2, 0.04 and 0.18 g per gram dry matter, indicating preferable hydration of pectin compared to cellulose. The effects of hydration are most noticeable in the measurements that reflect low frequency motions, particularly transverse relaxation, where both second moments and the relative intensity of signals arising from immobile material are reduced by hydration.

  8. Solid-state (13)C NMR and synchrotron SAXS/WAXS studies of uniaxially-oriented polyethylene.

    PubMed

    Afeworki, Mobae; Brant, Pat; Lustiger, Arnold; Norman, Alexander

    2015-11-01

    We report solid-state (13)C NMR and synchrotron wide-and small-angle X-ray scattering experiments (WAXS, SAXS) on metallocene linear low density polyethylene films (e.g., Exceed™ 1018 mLLDPE; nominally 1MI, 0.918 density ethylene-hexene metallocene copolymer) as a function of uniaxial draw ratio, λ. Combined, these experiments provide an unambiguous, quantitative molecular view of the orientation of both the crystalline and amorphous phases in the samples as a function of draw. Together with previously reported differential scanning calorimetry (DSC), gas transport measurements, transmission electron microscopy (TEM), optical birefringence, small angle X-ray scattering (SAXS) as well as other characterization techniques, this study of the state of orientation in both phases provides insight concerning the development of unusually high barrier properties of the most oriented samples (λ=10). In this work, static (non-spinning) solid-state NMR measurements indicate that in the drawn Exceed(TM) films both the crystalline and amorphous regions are highly oriented. In particular, chemical shift data show the amorphous phase is comprised increasingly of so-called "taut tie chains" (or tie chains under any state of tautness) in the mLLDPE with increasing draw ratio - the resonance lines associated with the amorphous phase shift to where the crystalline peaks are observed. In the sample with highest total draw (λ=10), virtually all of the chains in the non-crystalline region have responded and aligned in the machine (draw) direction. Both monoclinic and orthorhombic crystalline peaks are observed in high-resolution, solid-state magic-angle spinning (MAS) NMR measurements of the oriented PE films. The orientation is comparable to that obtained for ultra-high molecular weight HDPE fibers described as "ultra-oriented" in the literature. Furthermore, the presence of a monoclinic peak in cold-drawn samples suggests that there is an appreciable internal stress associated with the LLDPE. The results are confirmed and independently quantified by Herman's Orientation Function values derived from the WAXS measurements. The degree of orientation approaches theoretically perfect alignment of chains along the draw direction. We deduce from this observation that a high fraction of the non-crystalline chains are either tie chains that directly connect adjacent lamellae or are interlocking loops from adjacent lamellae. In either case, the chains are load-bearing and are consistent with the idea of "taut tie chains". We note that transmission electron micrographs recorded for the ultra-oriented Exceed showed the lamellae are often appreciably thinner and shorter than they are for cast or blown Exceed 1018. Combined with higher crystallinity, the thinner lamellae statistically favor more tie chains. Finally, the remarkably large decrease in permeability of the λ=10 film is primarily attributed to the high degree of orientation (and loss of entropy) of the amorphous phase. PMID:26476811

  9. Centrosymmetric silicide ScNiSi 3 - Structure, chemical bonding, and 45Sc solid state NMR

    NASA Astrophysics Data System (ADS)

    Harmening, Thomas; Eckert, Hellmut; Johrendt, Dirk; Pöttgen, Rainer

    2008-05-01

    The SmNiGe 3 type silicide ScNiSi 3, previously reported to crystallize in the non-centrosymmetric space group Amm2, was synthesized from the elements by arc-melting. Two different single crystals were investigated on the basis of X-ray diffractometer data: a = 382.0(1), b = 2058.2(1), c = 381.0(1) pm, wR2 = 0.0531, 947 F2 values for ScNi 0.913(2)Si 3 and a = 382.5(1), b = 2058.7(1), c = 380.8(1) pm, wR2 = 0.0494, 554 F2 values for ScNi 0.929(4)Si 3 with 23 variable parameters per refinement. The single crystal data unambiguously confirm the centrosymmetric space group Cmmm. The structure consists of a three-dimensional [NiSi 3] polyanion in which the scandium atoms fill distorted hexagonal channels. The polyanion contains three crystallographically independent silicon sites which are all involved in Si-Si bonding with Si-Si distances ranging from 232 to 235 pm. Each nickel atom has a square-pyramidal silicon coordination with rather short Ni-Si distances (217-222 pm). The nickel sites of both crystals show significant underfilling. The 45Sc magic-angle spinning NMR spectrum reveals the presence of a single scandium site, thereby confirming the centrosymmetric structure. In addition, a second, weaker resonance is attributed to scandium nuclei in the vicinity of a nickel vacancy. The fractional area of this resonance is quantitatively consistent with the nickel site occupancy of 92% determined from single crystal X-ray diffraction. Electronic structure calculations reveal metallic behaviour with strong Ni-Si and Si-Si bonds as expected, but additionally significant Sc-Ni bonding in contrast to YNiSi 3, where Y-Ni bonding is reported to be absent.

  10. Probing porous media with gas diffusion NMR

    NASA Technical Reports Server (NTRS)

    Mair, R. W.; Wong, G. P.; Hoffmann, D.; Hurlimann, M. D.; Patz, S.; Schwartz, L. M.; Walsworth, R. L.

    1999-01-01

    We show that gas diffusion nuclear magnetic resonance (GD-NMR) provides a powerful technique for probing the structure of porous media. In random packs of glass beads, using both laser-polarized and thermally polarized xenon gas, we find that GD-NMR can accurately measure the pore space surface-area-to-volume ratio, S/V rho, and the tortuosity, alpha (the latter quantity being directly related to the system's transport properties). We also show that GD-NMR provides a good measure of the tortuosity of sandstone and complex carbonate rocks.

  11. Closed-cycle cold helium magic-angle spinning for sensitivity-enhanced multi-dimensional solid-state NMR

    NASA Astrophysics Data System (ADS)

    Matsuki, Yoh; Nakamura, Shinji; Fukui, Shigeo; Suematsu, Hiroto; Fujiwara, Toshimichi

    2015-10-01

    Magic-angle spinning (MAS) NMR is a powerful tool for studying molecular structure and dynamics, but suffers from its low sensitivity. Here, we developed a novel helium-cooling MAS NMR probe system adopting a closed-loop gas recirculation mechanism. In addition to the sensitivity gain due to low temperature, the present system has enabled highly stable MAS (vR = 4-12 kHz) at cryogenic temperatures (T = 35-120 K) for over a week without consuming helium at a cost for electricity of 16 kW/h. High-resolution 1D and 2D data were recorded for a crystalline tri-peptide sample at T = 40 K and B0 = 16.4 T, where an order of magnitude of sensitivity gain was demonstrated versus room temperature measurement. The low-cost and long-term stable MAS strongly promotes broader application of the brute-force sensitivity-enhanced multi-dimensional MAS NMR, as well as dynamic nuclear polarization (DNP)-enhanced NMR in a temperature range lower than 100 K.

  12. Integrating Solid-State NMR and Computational Modeling to Investigate the Structure and Dynamics of Membrane-Associated Ghrelin

    PubMed Central

    Els-Heindl, Sylvia; Chollet, Constance; Scheidt, Holger A.; Beck-Sickinger, Annette G.; Meiler, Jens; Huster, Daniel

    2015-01-01

    The peptide hormone ghrelin activates the growth hormone secretagogue receptor 1a, also known as the ghrelin receptor. This 28-residue peptide is acylated at Ser3 and is the only peptide hormone in the human body that is lipid-modified by an octanoyl group. Little is known about the structure and dynamics of membrane-associated ghrelin. We carried out solid-state NMR studies of ghrelin in lipid vesicles, followed by computational modeling of the peptide using Rosetta. Isotropic chemical shift data of isotopically labeled ghrelin provide information about the peptide’s secondary structure. Spin diffusion experiments indicate that ghrelin binds to membranes via its lipidated Ser3. Further, Phe4, as well as electrostatics involving the peptide’s positively charged residues and lipid polar headgroups, contribute to the binding energy. Other than the lipid anchor, ghrelin is highly flexible and mobile at the membrane surface. This observation is supported by our predicted model ensemble, which is in good agreement with experimentally determined chemical shifts. In the final ensemble of models, residues 8–17 form an α-helix, while residues 21–23 and 26–27 often adopt a polyproline II helical conformation. These helices appear to assist the peptide in forming an amphipathic conformation so that it can bind to the membrane. PMID:25803439

  13. Integrating solid-state NMR and computational modeling to investigate the structure and dynamics of membrane-associated ghrelin.

    PubMed

    Vortmeier, Gerrit; DeLuca, Stephanie H; Els-Heindl, Sylvia; Chollet, Constance; Scheidt, Holger A; Beck-Sickinger, Annette G; Meiler, Jens; Huster, Daniel

    2015-01-01

    The peptide hormone ghrelin activates the growth hormone secretagogue receptor 1a, also known as the ghrelin receptor. This 28-residue peptide is acylated at Ser3 and is the only peptide hormone in the human body that is lipid-modified by an octanoyl group. Little is known about the structure and dynamics of membrane-associated ghrelin. We carried out solid-state NMR studies of ghrelin in lipid vesicles, followed by computational modeling of the peptide using Rosetta. Isotropic chemical shift data of isotopically labeled ghrelin provide information about the peptide's secondary structure. Spin diffusion experiments indicate that ghrelin binds to membranes via its lipidated Ser3. Further, Phe4, as well as electrostatics involving the peptide's positively charged residues and lipid polar headgroups, contribute to the binding energy. Other than the lipid anchor, ghrelin is highly flexible and mobile at the membrane surface. This observation is supported by our predicted model ensemble, which is in good agreement with experimentally determined chemical shifts. In the final ensemble of models, residues 8-17 form an α-helix, while residues 21-23 and 26-27 often adopt a polyproline II helical conformation. These helices appear to assist the peptide in forming an amphipathic conformation so that it can bind to the membrane. PMID:25803439

  14. Structural topology of phospholamban pentamer in lipid bilayers by a hybrid solution and solid-state NMR method

    PubMed Central

    Verardi, Raffaello; Shi, Lei; Traaseth, Nathaniel J.; Walsh, Naomi; Veglia, Gianluigi

    2011-01-01

    Phospholamban (PLN) is a type II membrane protein that inhibits the sarcoplasmic reticulum Ca2+-ATPase (SERCA), thereby regulating calcium homeostasis in cardiac muscle. In membranes, PLN forms pentamers that have been proposed to function either as a storage for active monomers or as ion channels. Here, we report the T-state structure of pentameric PLN solved by a hybrid solution and solid-state NMR method. In lipid bilayers, PLN adopts a pinwheel topology with a narrow hydrophobic pore, which excludes ion transport. In the T state, the cytoplasmic amphipathic helices (domains Ia) are absorbed into the lipid bilayer with the transmembrane domains arranged in a left-handed coiled-coil configuration, crossing the bilayer with a tilt angle of approximately 11° with respect to the membrane normal. The tilt angle difference between the monomer and pentamer is approximately 13°, showing that intramembrane helix–helix association forces dominate over the hydrophobic mismatch, driving the overall topology of the transmembrane assembly. Our data reveal that both topology and function of PLN are shaped by the interactions with lipids, which fine-tune the regulation of SERCA. PMID:21576492

  15. Advanced solid-state NMR techniques for characterization of membrane protein structure and dynamics: application to Anabaena Sensory Rhodopsin.

    PubMed

    Ward, Meaghan E; Brown, Leonid S; Ladizhansky, Vladimir

    2015-04-01

    Studies of the structure, dynamics, and function of membrane proteins (MPs) have long been considered one of the main applications of solid-state NMR (SSNMR). Advances in instrumentation, and the plethora of new SSNMR methodologies developed over the past decade have resulted in a number of high-resolution structures and structural models of both bitopic and polytopic α-helical MPs. The necessity to retain lipids in the sample, the high proportion of one type of secondary structure, differential dynamics, and the possibility of local disorder in the loop regions all create challenges for structure determination. In this Perspective article we describe our recent efforts directed at determining the structure and functional dynamics of Anabaena Sensory Rhodopsin, a heptahelical transmembrane (7TM) protein. We review some of the established and emerging methods which can be utilized for SSNMR-based structure determination, with a particular focus on those used for ASR, a bacterial protein which shares its 7TM architecture with G-protein coupled receptors. PMID:25637099

  16. Solid-state NMR structure of a pathogenic fibril of full-length human α-synuclein.

    PubMed

    Tuttle, Marcus D; Comellas, Gemma; Nieuwkoop, Andrew J; Covell, Dustin J; Berthold, Deborah A; Kloepper, Kathryn D; Courtney, Joseph M; Kim, Jae K; Barclay, Alexander M; Kendall, Amy; Wan, William; Stubbs, Gerald; Schwieters, Charles D; Lee, Virginia M Y; George, Julia M; Rienstra, Chad M

    2016-05-01

    Misfolded α-synuclein amyloid fibrils are the principal components of Lewy bodies and neurites, hallmarks of Parkinson's disease (PD). We present a high-resolution structure of an α-synuclein fibril, in a form that induces robust pathology in primary neuronal culture, determined by solid-state NMR spectroscopy and validated by EM and X-ray fiber diffraction. Over 200 unique long-range distance restraints define a consensus structure with common amyloid features including parallel, in-register β-sheets and hydrophobic-core residues, and with substantial complexity arising from diverse structural features including an intermolecular salt bridge, a glutamine ladder, close backbone interactions involving small residues, and several steric zippers stabilizing a new orthogonal Greek-key topology. These characteristics contribute to the robust propagation of this fibril form, as supported by the structural similarity of early-onset-PD mutants. The structure provides a framework for understanding the interactions of α-synuclein with other proteins and small molecules, to aid in PD diagnosis and treatment. PMID:27018801

  17. Solid-State (13)C NMR Delineates the Architectural Design of Biopolymers in Native and Genetically Altered Tomato Fruit Cuticles.

    PubMed

    Chatterjee, Subhasish; Matas, Antonio J; Isaacson, Tal; Kehlet, Cindie; Rose, Jocelyn K C; Stark, Ruth E

    2016-01-11

    Plant cuticles on outer fruit and leaf surfaces are natural macromolecular composites of waxes and polyesters that ensure mechanical integrity and mitigate environmental challenges. They also provide renewable raw materials for cosmetics, packaging, and coatings. To delineate the structural framework and flexibility underlying the versatile functions of cutin biopolymers associated with polysaccharide-rich cell-wall matrices, solid-state NMR spectra and spin relaxation times were measured in a tomato fruit model system, including different developmental stages and surface phenotypes. The hydrophilic-hydrophobic balance of the cutin ensures compatibility with the underlying polysaccharide cell walls; the hydroxy fatty acid structures of outer epidermal cutin also support deposition of hydrophobic waxes and aromatic moieties while promoting the formation of cell-wall cross-links that rigidify and strengthen the cuticle composite during fruit development. Fruit cutin-deficient tomato mutants with compromised microbial resistance exhibit less efficient local and collective biopolymer motions, stiffening their cuticular surfaces and increasing their susceptibility to fracture. PMID:26652188

  18. Investigation of drug-excipient interactions in lapatinib amorphous solid dispersions using solid-state NMR spectroscopy.

    PubMed

    Song, Yang; Yang, Xinghao; Chen, Xin; Nie, Haichen; Byrn, Stephen; Lubach, Joseph W

    2015-03-01

    This study investigated the presence of specific drug-excipient interactions in amorphous solid dispersions of lapatinib (LB) and four commonly used pharmaceutical polymers, including Soluplus, polyvinylpyrrolidone vinyl acetate (PVPVA), hydroxypropylmethylcellulose acetate succinate (HPMCAS), and hydroxypropylmethylcellulose phthalate (HPMCP). Based on predicted pKa differences, LB was hypothesized to exhibit a specific ionic interaction with HPMCP, and possibly with HPMCAS, while Soluplus and PVPVA were studied as controls without ionizable functionality. Thermal studies showed a single glass transition (Tg) for each dispersion, in close agreement with predicted values for Soluplus, PVPVA, and HPMCAS systems. However, the Tg values of LB-HPMCP solid dispersions were markedly higher than predicted values, indicating a strong intermolecular interaction between LB and HPMCP. (15)N solid-state NMR provided direct spectroscopic evidence for protonation of LB (i.e., salt formation) within the HPMCP solid dispersions. (1)H T1 and (1)H T1ρ relaxation studies of the dispersions supported the ionic interaction hypothesis, and indicated multiple phases in the cases of excess drug or polymer. In addition, the dissolution and stability behavior of each system was examined. Both acidic polymers, HPMCAS and HPMCP, effectively inhibited the crystallization of LB on accelerated stability, likely owing to beneficial strong intermolecular hydrogen and/or specific ionic bonds with the acidic polymers. Soluplus and PVPVA showed poor physical properties on stability and subsequently poor crystallization inhibition. PMID:25585133

  19. Structural Insight into an Alzheimer’s Brain-Derived Spherical Assembly of Amyloid β by Solid-State NMR

    PubMed Central

    2016-01-01

    Accumulating evidence suggests that various neurodegenerative diseases, including Alzheimer’s disease (AD), are linked to cytotoxic diffusible aggregates of amyloid proteins, which are metastable intermediate species in protein misfolding. This study presents the first site-specific structural study on an intermediate called amylospheroid (ASPD), an AD-derived neurotoxin composed of oligomeric amyloid-β (Aβ). Electron microscopy and immunological analyses using ASPD-specific “conformational” antibodies established synthetic ASPD for the 42-residue Aβ(1–42) as an excellent structural/morphological analogue of native ASPD extracted from AD patients, the level of which correlates with the severity of AD. 13C solid-state NMR analyses of approximately 20 residues and interstrand distances demonstrated that the synthetic ASPD is made of a homogeneous single conformer containing parallel β-sheets. These results provide profound insight into the native ASPD, indicating that Aβ is likely to self-assemble into the toxic intermediate with β-sheet structures in AD brains. This approach can be applied to various intermediates relevant to amyloid diseases. PMID:25938164

  20. Characterization of phosphate sequestration by a lanthanum modified bentonite clay: A solid- state NMR, EXAFS and PXRD study

    SciTech Connect

    Dithmer, Line; Lipton, Andrew S.; Reitzel, Kasper; Warner, Terence E.; Lundberg, Daniel; Nielsen, Ulla Gro

    2015-04-07

    Phosphate (P) sequestration by a lanthanum (La) exchanged bentonite (a clay mineral), which is extensively used in chemical lake restoration, was investigated on the molecular level using a combination of 31P and 139La solid state NMR spectroscopy (SSNMR), extended X-ray absorption spectroscopy (EX-AFS) and powder X-ray diffraction (PXRD) in combination with sorption studies. 31P SSNMR show that all phosphate is immobilized as rhabdophane, LaPO4·xH2O, which is further supported by 139La SSNMR and EXAFS; whereas PXRD results are ambiguous with respect to rhabdophane and monazite (LaPO4). Adsorption studies show that, at humic acids (HA) concentrations above ca. 250 μM the binding capacity is only 50 % of the theoretical value or even less. No other lanthanum or phosphate phases are detected by SSNMR and EXAFS indicating the effect of HA is kinetic. Moreover, 31P SSNMR shows that rhabdophane formed upon P sequestration is in close proximity to the clay matrix.

  1. Characterization of phosphate sequestration by a lanthanum modified bentonite clay: a solid-state NMR, EXAFS, and PXRD study.

    PubMed

    Dithmer, Line; Lipton, Andrew S; Reitzel, Kasper; Warner, Terence E; Lundberg, Daniel; Nielsen, Ulla Gro

    2015-04-01

    Phosphate (Pi) sequestration by a lanthanum (La) exchanged clay mineral (La-Bentonite), which is extensively used in chemical lake restoration, was investigated on the molecular level using a combination of (31)P and (139)La solid state NMR spectroscopy (SSNMR), extended X-ray absorption spectroscopy (EXAFS), powder X-ray diffraction (PXRD) and sorption studies. (31)P SSNMR show that all Pi was immobilized as rhabdophane (LaPO4·n H2O, n ≤ 3), which was further supported by (139)La SSNMR and EXAFS. However, PXRD results were ambiguous with respect to rhabdophane and monazite (LaPO4). Adsorption studies showed that at dissolved organic carbon (DOC) concentration above ca. 250 μM the binding capacity was only 50% of the theoretical value or even less. No other La or Pi phases were detected by SSNMR and EXAFS indicating the effect of DOC is kinetic. Moreover, (31)P SSNMR showed that rhabdophane formed upon Pi sequestration is in close proximity to the clay matrix. PMID:25747941

  2. Spin dynamics with Solid State NMR and GPU calculations: Loschmidt Echoes, Intrinsic Decoherence and Quantum Dynamical Phase Transitions

    NASA Astrophysics Data System (ADS)

    Pastawski, Horacio M.

    2015-03-01

    After overviewing argentine Condensed Matter Physics outside the Metropolitan area I will focus on the Loschmidt Echo [LE], a concept developed and pursed at Córdoba. It is the recovered fraction of a localized excitation after a spreading period followed by an imperfect time reversal procedure. In Solid State NMR, the LE has allowed us to quantify the decoherence and irreversibility induced by an uncontrolled environment. Notably complex many-body dynamics makes the system particularly sensitive to environmental disturbances presenting a decoherence rate that becomes perturbation independent beyond some small threshold. These experiments and the theoretical analysis based on the Feynman's path integral, summarized at a tutorial level, fueled the field of dynamical quantum chaos [4]. The quest for a perturbation independent decoherence as an emergent phenomenon in thermodynamic limit, lead us to discuss other dynamical observables that depend non-analytically on the environment strength, i.e. that undergo a quantum dynamical phase transition QDPT. GPU based high performance computing boosts the evaluation of the LE, allowing us to asses thermalization and how the Metal-Insulator transition (also a QDPT) emerges in interacting many-body systems. Financing institutions: CONICET, SeCyT-UNC, ANPCyT, MinCyT-Cor and Antorchas.

  3. Solid-State Selective 13C Excitation and Spin Diffusion NMR to Resolve Spatial Dimensions in Plant Cell Walls

    SciTech Connect

    Foston, M.; Katahira, R.; Gjersing, E.; Davis, M. F.; Ragauskas, A. J.

    2012-02-15

    The average spatial dimensions between major biopolymers within the plant cell wall can be resolved using a solid-state NMR technique referred to as a {sup 13}C cross-polarization (CP) SELDOM (selectively by destruction of magnetization) with a mixing time delay for spin diffusion. Selective excitation of specific aromatic lignin carbons indicates that lignin is in close proximity to hemicellulose followed by amorphous and finally crystalline cellulose. {sup 13}C spin diffusion time constants (T{sub SD}) were extracted using a two-site spin diffusion theory developed for {sup 13}C nuclei under magic angle spinning (MAS) conditions. These time constants were then used to calculate an average lower-limit spin diffusion length between chemical groups within the plant cell wall. The results on untreated {sup 13}C enriched corn stover stem reveal that the lignin carbons are, on average, located at distances {approx}0.7-2.0 nm from the carbons in hemicellulose and cellulose, whereas the pretreated material had larger separations.

  4. Crystal Polymorphism of Protein GB1 Examined by Solid-State NMR Spectroscopy and X-ray Diffraction

    PubMed Central

    Frericks Schmidt, Heather L.; Sperling, Lindsay J.; Gao, Yi Gui; Wylie, Benjamin J.; Boettcher, John M.; Wilson, Scott R.; Rienstra, Chad M.

    2009-01-01

    The study of micro- or nanocrystalline proteins by magic-angle spinning (MAS) solid-state NMR (SSNMR) gives atomic-resolution insight into structure in cases when single crystals cannot be obtained for diffraction studies. Subtle differences in the local chemical environment around the protein, including the characteristics of the co-solvent and the buffer, determine whether a protein will form single crystals. The impact of these small changes in formulation is also evident in the SSNMR spectra, but leads only to correspondingly subtle changes in the spectra. Here we demonstrate that several formulations of GB1 microcrystals yield very high-quality SSNMR spectra, although only a subset of conditions enable growth of single crystals. We have characterized these polymorphs by X-ray powder diffraction and assigned the SSNMR spectra. Assignments of the 13C and 15N SSNMR chemical shifts confirm that the backbone structure is conserved, indicative of a common protein fold, but sidechain chemical shifts are changed on the surface of the protein, in a manner dependent upon crystal packing and electrostatic interactions with salt in the mother liquor. Our results demonstrate the ability of SSNMR to reveal minor structural differences among crystal polymorphs. This ability has potential practical utility for studying formulation chemistry of industrial and therapeutic proteins, as well as for deriving fundamental insights into the phenomenon of single crystal growth. PMID:18052145

  5. Comparison of celery (Apium graveolens L.) collenchyma and parenchyma cell wall polysaccharides enabled by solid-state (13)C NMR.

    PubMed

    Zujovic, Zoran; Chen, Da; Melton, Laurence D

    2016-02-01

    Collenchyma cells with their thickened walls are one of specific mechanical support tissues for plants, while parenchyma cells are thin walled and serve multiple functions. The parenchyma tissue is what you enjoy eating, while collenchyma, because of its fibrous nature, is not so attractive. Celery is a useful model for comparing the cell walls (CWs) of the two cell types such as collenchyma and parenchyma. However, to date, the structural characteristics of collenchyma and parenchyma cell walls from the same plant have not been compared. Monosaccharide composition suggested the collenchyma cell walls contained less pectin but more hemicellulose in comparison to parenchyma. High-resolution solid-state NMR spectra of highly mobile pectins revealed that the arabinan signals were more evident in the collenchyma spectrum, while galactan showed a much stronger resonance in the parenchyma spectrum. In addition, methyl esterified and non-esterified galacturonic acid signals were observed in parenchyma CWs, but only the latter one appeared in the collenchyma. The ratio of cellulose surface/interior obtained from CP/MAS spectra for collenchyma suggested the cellulose microfibrils were ~2.4 nm, while in the parenchyma, these were somewhat larger. X-ray diffraction indicated the size of the cellulose microfibrils were the same for both types of CWs. PMID:26717549

  6. Static solid-state (14)N NMR and computational studies of nitrogen EFG tensors in some crystalline amino acids.

    PubMed

    O'Dell, Luke A; Schurko, Robert W

    2009-08-28

    The recently reported direct enhancement of integer spin magnetization (DEISM) methodology for signal enhancement in solid-state NMR of integer spins has been used to obtain static (14)N powder patterns from alpha-glycine, L-leucine and L-proline in relatively short experimental times at 9.4 T, allowing accurate determination of the quadrupolar parameters. Proton decoupling and deuteration of the nitrogen sites were used to reduce the (1)H-(14)N dipolar contribution to the transverse relaxation time allowing more echoes to be acquired per scan. In addition, ab initio calculations using molecular clusters (Gaussian 03) and the full crystal lattice (CASTEP) have been employed to confirm these results, to obtain the orientation of the electric field gradient (EFG) tensors in the molecular frame, and also to correctly assign the two sets of parameters for L-leucine. The (14)N EFG tensor is shown to be highly sensitive to the surrounding environment, particularly to nearby hydrogen bonding. PMID:19652842

  7. Study of the Beckmann rearrangement of acetophenone oxime over porous solids by means of solid state NMR spectroscopy.

    PubMed

    Fernandez, Ana Belen; Lezcano-Gonzalez, Ines; Boronat, Mercedes; Blasco, Teresa; Corma, Avelino

    2009-07-01

    The Beckmann rearrangement of acetophenone oxime using zeolite H-beta and silicalite-N as catalysts has been investigated by means of (15)N and (13)C solid state NMR spectroscopy in combination with theoretical calculations. The results obtained show that the oxime is N-protonated at room temperature on the acid sites of zeolite H-beta. At reaction temperatures of 423 K or above, the two isomeric amides, acetanilide and N-methyl benzamide (NMB) are formed, and interact with the Brønsted acid sites of zeolite H-beta through hydrogen bonds. The presence of residual water hydrolyzes the two amides, while larger amounts inhibit the formation of NMB and cause the total hydrolysis of the acetanilide. Over siliceous zeolite silicalite-N, containing silanol nests as active sites, the oxime is adsorbed through hydrogen bonds and only acetanilide is formed at reaction temperatures of 423 K or above. In the presence of water, the reaction starts at 473 K, still being very selective up to 573 K, and the amide is partially hydrolyzed only above this temperature . PMID:19562146

  8. Advanced solid-state NMR techniques for characterization of membrane protein structure and dynamics: Application to Anabaena Sensory Rhodopsin

    NASA Astrophysics Data System (ADS)

    Ward, Meaghan E.; Brown, Leonid S.; Ladizhansky, Vladimir

    2015-04-01

    Studies of the structure, dynamics, and function of membrane proteins (MPs) have long been considered one of the main applications of solid-state NMR (SSNMR). Advances in instrumentation, and the plethora of new SSNMR methodologies developed over the past decade have resulted in a number of high-resolution structures and structural models of both bitopic and polytopic α-helical MPs. The necessity to retain lipids in the sample, the high proportion of one type of secondary structure, differential dynamics, and the possibility of local disorder in the loop regions all create challenges for structure determination. In this Perspective article we describe our recent efforts directed at determining the structure and functional dynamics of Anabaena Sensory Rhodopsin, a heptahelical transmembrane (7TM) protein. We review some of the established and emerging methods which can be utilized for SSNMR-based structure determination, with a particular focus on those used for ASR, a bacterial protein which shares its 7TM architecture with G-protein coupled receptors.

  9. Differentiation of histidine tautomeric states using 15N selectively filtered 13C solid-state NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Miao, Yimin; Cross, Timothy A.; Fu, Riqiang

    2014-08-01

    The histidine imidazole ring in proteins usually contains a mixture of three possible tautomeric states (two neutral - ? and ? states and a charged state) at physiological pHs. Differentiating the tautomeric states is critical for understanding how the histidine residue participates in many structurally and functionally important proteins. In this work, one dimensional 15N selectively filtered 13C solid-state NMR spectroscopy is proposed to differentiate histidine tautomeric states and to identify all 13C resonances of the individual imidazole rings in a mixture of tautomeric states. When 15N selective 180 pulses are applied to the protonated or non-protonated nitrogen region, the 13C sites that are bonded to the non-protonated or protonated nitrogen sites can be identified, respectively. A sample of 13C, 15N labeled histidine powder lyophilized from a solution at pH 6.3 has been used to illustrate the usefulness of this scheme by uniquely assigning resonances of the neutral ? and charged states from the mixture.

  10. Crystal Polymorphism of Protein GB1 Examined by Solid-State NMR Spectroscopy and X-ray Diffraction

    SciTech Connect

    Schmidt,H.; Sperling, L.; Gao, Y.; Wylie, B.; Boettcher, J.; Wilson, S.; Rienstra, C.

    2007-01-01

    The study of micro- or nanocrystalline proteins by magic-angle spinning (MAS) solid-state NMR (SSNMR) gives atomic-resolution insight into structure in cases when single crystals cannot be obtained for diffraction studies. Subtle differences in the local chemical environment around the protein, including the characteristics of the cosolvent and the buffer, determine whether a protein will form single crystals. The impact of these small changes in formulation is also evident in the SSNMR spectra; however, the changes lead only to correspondingly subtle changes in the spectra. Here, we demonstrate that several formulations of GB1 microcrystals yield very high quality SSNMR spectra, although only a subset of conditions enable growth of single crystals. We have characterized these polymorphs by X-ray powder diffraction and assigned the SSNMR spectra. Assignments of the 13C and 15N SSNMR chemical shifts confirm that the backbone structure is conserved, indicative of a common protein fold, but side chain chemical shifts are changed on the surface of the protein, in a manner dependent upon crystal packing and electrostatic interactions with salt in the mother liquor. Our results demonstrate the ability of SSNMR to reveal minor structural differences among crystal polymorphs. This ability has potential practical utility for studying the formulation chemistry of industrial and therapeutic proteins, as well as for deriving fundamental insights into the phenomenon of single-crystal growth.

  11. The 31P solid state NMR studies of ZrP, Mg3P2, and CdPS3

    NASA Astrophysics Data System (ADS)

    Nissan, R. A.; Vanderah, T. A.

    The 31P solid state NMR spectra of ZrP, Mg3P2, MgP4, and CdPS3 are reported. Static and magic angle spinning (MAS) spectra were obtained for each compound. In all cases, chemical shift anisotropy and the effects of dipolar broadening were sufficiently reduced by the MAS method to reveal the isotropic chemical shifts for each crystallographically distinct phosphorus. The observed resonances were assigned to the different types of phosphorus by considering the structural details of each compound. In ZrP the two chemical shifts of +128.4 and +187.5 ppm (relative to 85 percent H3PO4) were assigned to P occupying the Wyckoff 2d and 2a sites, respectively. In Mg3P2, two resonances from the 24d and 8a site P atoms were observed at -262.3 and -239.6 ppm, respectively. In MgP4 two types of phosphorus, one type coordinated to two Mg and two P and the other to three P and one Mg, gave chemical shifts of -109.2 and -6.1 ppm, respectively. From CdPS3 only one resonance at +104.9 ppm is observed as all P atoms are crystallographically equivalent.

  12. Solid-state and solution /sup 13/C NMR in the conformational analysis of methadone-hydrochloride and related narcotic analgesics

    SciTech Connect

    Sumner, S.C.J.

    1986-01-01

    Solid state and solution /sup 13/C NMR have been used to study the conformations of the racemic mixtures and single enantiomers of methadone hydrochloride, alpha and beta methadol hydrochloride, and alpha and beta acetylmethadol hydrochloride. The NMR spectra acquired for the compounds as solids, and in polar and nonpolar solvents are compared, in order to determine the conformation of the molecules in solution. To determine the reliability of assigning solution conformations by comparing solution and solid state chemical shift data, three bond coupling constants measured in solution are compared with those calculated from X-ray data. The conformations of the racemic mixture and plus enantiomer of methadone hydrochloride have been shown to be very similar in the solid state, where minor differences in conformation can be seen by comparing NMR spectra obtained for the solids. Also shown is that the molecules of methadone hydrochloride have conformations in polar and in nonpolar solvents which are very similar to the conformation of the molecules in the solid state.

  13. New opportunities for high-resolution solid-state NMR spectroscopy of oxide materials at 21.1- and 18.8-T fields.

    PubMed

    Stebbins, J; Du, Lin-Shu; Kroeker, S; Neuhoff, P; Rice, D; Frye, J; Jakobsen, H J

    2002-01-01

    We present here high-resolution solid state NMR spectra of several oxide and silicate materials that illustrate the improvements obtainable with very high external fields (18.8 and 21.1 T), with probes capable of tuning to a wide frequency range that allow observations of nuclides from high to low magnetogyric ratio. We discuss 27Al MAS spectra for the zeolite scolecite (CaAl2Si3O10 x 3H2O), 17O MAS data for analcime (NaAlSi2O6 x H2O), calcium monoaluminate (CaAI2O4), and titanite (CaTiSiO5), 39K spin-echo spectra for leucite (KAlSi2O6), microline (KAlSiO8), muscovite (KAl2(AlSi3O10)(OH2) and a potassium aluminosolicate glass, and preliminary 73Ge spin-echo MAS spectra for crystalline and glassy germanium dioxide (GeO2). PMID:11949814

  14. Glassy carbons from poly(furfuryl alcohol) copolymers: structural studies by high-resolution solid-state NMR techniques

    SciTech Connect

    Eckert, H.; Levendis, Y.A.; Flagan, R.C.

    1988-08-25

    The chemical structure of glass carbon particles produced from poly(furfuryl alcohol) copolymers is studied by /sup 13/C cross-polarization/magic-angle spinning (CP-MAS) NMR and high-speed /sup 1/H MAS NMR. In agreement with earlier proposals, /sup 13/C NMR spectra confirm the buildup of a highly unsaturated system at the expense of furan rings and aliphatic carbon atoms, and upon heating to 800 K this conversion is essentially complete. Successive carbonization by air oxidation or pyrolysis at temperatures up to 1600 K is reflected in a gradual decrease of the /sup 13/C chemical shift from ca. 130 to 115 ppm versus tetramethylsilane. /sup 1/H MAS NMR is used to detect and quantitate the amount of residual C-bonded hydrogen species at various stages of the carbonization process. In addition, these spectra show intense, narrow resonances due to sorbed H/sub 2/O molecules, which resonate over a wide range of chemical shifts (between 2.5 and /minus/8 ppm versus tetramethylsilane). In analogy with effects observed by Tabony and co-workers for molecules adsorbed above the basal plane of graphite, the upfield shifts observed for water sorbed in the glassy carbons of the present study are attributed to the large susceptibility anisotropy of submicroscopically ordered, turbostratic, or partially graphitized regions of the samples. The extent of this ordering is inversely correlated with the absolute content of residual C-bonded hydrogen species and depends mainly on the temperature of pyrolysis, whereas the oxygen content of the heating atmosphere and the composition of the initial polymeric material appear to be of secondary importance. The results suggest that sorbed H/sub 2/O molecules can function as sensitive NMR chemical shift probes for the initial stages of crystallization processes in glassy carbons.

  15. Sensitivity and resolution enhanced solid-state NMR for paramagnetic systems and biomolecules under very fast magic angle spinning.

    PubMed

    Parthasarathy, Sudhakar; Nishiyama, Yusuke; Ishii, Yoshitaka

    2013-09-17

    Recent research in fast magic angle spinning (MAS) methods has drastically improved the resolution and sensitivity of NMR spectroscopy of biomolecules and materials in solids. In this Account, we summarize recent and ongoing developments in this area by presenting (13)C and (1)H solid-state NMR (SSNMR) studies on paramagnetic systems and biomolecules under fast MAS from our laboratories. First, we describe how very fast MAS (VFMAS) at the spinning speed of at least 20 kHz allows us to overcome major difficulties in (1)H and (13)C high-resolution SSNMR of paramagnetic systems. As a result, we can enhance both sensitivity and resolution by up to a few orders of magnitude. Using fast recycling (?ms/scan) with short (1)H T1 values, we can perform (1)H SSNMR microanalysis of paramagnetic systems on the microgram scale with greatly improved sensitivity over that observed for diamagnetic systems. Second, we discuss how VFMAS at a spinning speed greater than ?40 kHz can enhance the sensitivity and resolution of (13)C biomolecular SSNMR measurements. Low-power (1)H decoupling schemes under VFMAS offer excellent spectral resolution for (13)C SSNMR by nominal (1)H RF irradiation at ?10 kHz. By combining the VFMAS approach with enhanced (1)H T1 relaxation by paramagnetic doping, we can achieve extremely fast recycling in modern biomolecular SSNMR experiments. Experiments with (13)C-labeled ubiquitin doped with 10 mM Cu-EDTA demonstrate how effectively this new approach, called paramagnetic assisted condensed data collection (PACC), enhances the sensitivity. Lastly, we examine (13)C SSNMR measurements for biomolecules under faster MAS at a higher field. Our preliminary (13)C SSNMR data of A? amyloid fibrils and GB1 microcrystals acquired at (1)H NMR frequencies of 750-800 MHz suggest that the combined use of the PACC approach and ultrahigh fields could allow for routine multidimensional SSNMR analyses of proteins at the 50-200 nmol level. Also, we briefly discuss the prospects for studying bimolecules using (13)C SSNMR under ultrafast MAS at the spinning speed of ?100 kHz. PMID:23889329

  16. Comprehensive signal assignment of 13C-labeled lignocellulose using multidimensional solution NMR and 13C chemical shift comparison with solid-state NMR.

    PubMed

    Komatsu, Takanori; Kikuchi, Jun

    2013-09-17

    A multidimensional solution NMR method has been developed using various pulse programs including HCCH-COSY and (13)C-HSQC-NOESY for the structural characterization of commercially available (13)C labeled lignocellulose from potatoes (Solanum tuberosum L.), chicory (Cichorium intybus), and corn (Zea mays). This new method allowed for 119 of the signals in the (13)C-HSQC spectrum of lignocelluloses to be assigned and was successfully used to characterize the structures of lignocellulose samples from three plants in terms of their xylan and xyloglucan structures, which are the major hemicelluloses in angiosperm. Furthermore, this new method provided greater insight into fine structures of lignin by providing a high resolution to the aromatic signals of the β-aryl ether and resinol moieties, as well as the diastereomeric signals of the β-aryl ether. Finally, the (13)C chemical shifts assigned in this study were compared with those from solid-state NMR and indicated the presence of heterogeneous dynamics in the polysaccharides where rigid cellulose and mobile hemicelluloses moieties existed together. PMID:24010724

  17. Solid-state NMR adiabatic TOBSY sequences provide enhanced sensitivity for multidimensional high-resolution magic-angle-spinning 1H MR spectroscopy.

    PubMed

    Andronesi, Ovidiu C; Mintzopoulos, Dionyssios; Struppe, Jochem; Black, Peter M; Tzika, A Aria

    2008-08-01

    We propose a solid-state NMR method that maximizes the advantages of high-resolution magic-angle-spinning (HRMAS) applied to intact biopsies when compared to more conventional liquid-state NMR approaches. Theoretical treatment, numerical simulations and experimental results on intact human brain biopsies are presented. Experimentally, it is proven that an optimized adiabatic TOBSY (TOtal through Bond correlation SpectroscopY) solid-state NMR pulse sequence for two-dimensional 1H-1H homonuclear scalar-coupling longitudinal isotropic mixing provides a 20%-50% improvement in signal-to-noise ratio relative to its liquid-state analogue TOCSY (TOtal Correlation SpectroscopY). For this purpose we have refined the C9(15)1 symmetry-based 13C TOBSY pulse sequence for 1H MRS use and compared it to MLEV-16 TOCSY sequence. Both sequences were rotor-synchronized and implemented using WURST-8 adiabatic inversion pulses. As discussed theoretically and shown in simulations, the improved magnetization-transfer comes from actively removing residual dipolar couplings from the average Hamiltonian. Importantly, the solid-state NMR techniques are tailored to perform measurements at low temperatures where sample degradation is reduced. This is the first demonstration of such a concept for HRMAS metabolic profiling of disease processes, including cancer, from biopsies requiring reduced sample degradation for further genomic analysis. PMID:18556227

  18. 33S NMR cryogenic probe for taurine detection

    NASA Astrophysics Data System (ADS)

    Hobo, Fumio; Takahashi, Masato; Maeda, Hideaki

    2009-03-01

    With the goal of a S33 nuclear magnetic resonance (NMR) probe applicable to in vivo NMR on taurine-biological samples, we have developed the S33 NMR cryogenic probe, which is applicable to taurine solutions. The NMR sensitivity gain relative to a conventional broadband probe is as large as 3.5. This work suggests that improvements in the preamplifier could allow NMR measurements on 100 μM taurine solutions, which is the level of sensitivity necessary for biological samples.

  19. (51)V solid-state NMR and density functional theory studies of eight-coordinate non-oxo vanadium complexes: oxidized amavadin.

    PubMed

    Ooms, Kristopher J; Bolte, Stephanie E; Baruah, Bharat; Choudhary, Muhammad Aziz; Crans, Debbie C; Polenova, Tatyana

    2009-05-01

    Using (51)V magic angle spinning solid-state NMR spectroscopy and density functional theory calculations we have characterized the chemical shift and quadrupolar coupling parameters for two eight-coordinate vanadium complexes, [PPh(4)][V(v)(HIDPA)(2)] and [PPh(4)][V(v)(HIDA)(2)]; HIDPA = 2,2'-(hydroxyimino)dipropionate and HIDA = 2,2'-(hydroxyimino)diacetate. The coordination geometry under examination is the less common non-oxo eight coordinate distorted dodecahedral geometry that has not been previously investigated by solid-state NMR spectroscopy. Both complexes were isolated by oxidizing their reduced forms: [V(iv)(HIDPA)(2)](2-) and [V(iv)(HIDA)(2)](2-). V(iv)(HIDPA)(2)(2-) is also known as amavadin, a vanadium-containing natural product present in the Amanita muscaria mushroom and is responsible for vanadium accumulation in nature. The quadrupolar coupling constants, C(Q), are found to be moderate, 5.0-6.4 MHz while the chemical shift anisotropies are relatively small for vanadium complexes, -420 and -360 ppm. The isotropic chemical shifts in the solid state are -220 and -228 ppm for the two compounds, and near the chemical shifts observed in solution. Presumably this is a consequence of the combined effects of the increased coordination number and the absence of oxo groups. Density functional theory calculations of the electric field gradient parameters are in good agreement with the NMR results while the chemical shift parameters show some deviation from the experimental values. Future work on this unusual coordination geometry and a combined analysis by solid-state NMR and density functional theory should provide a better understanding of the correlations between experimental NMR parameters and the local structure of the vanadium centers. PMID:19421628

  20. Ca(2+) ATPase Conformational Transitions in Lipid Bilayers Mapped by Site-directed Ethylation and Solid-State NMR.

    PubMed

    Vostrikov, Vitaly V; Gustavsson, Martin; Gopinath, Tata; Mullen, Dan; Dicke, Alysha A; Truong, Vincent; Veglia, Gianluigi

    2016-02-19

    To transmit signals across cellular compartments, many membrane-embedded enzymes undergo extensive conformational rearrangements. Monitoring these events in lipid bilayers by NMR at atomic resolution has been challenging due to the large size of these systems. It is further exacerbated for large mammalian proteins that are difficult to express and label with NMR-active isotopes. Here, we synthesized and engineered (13)C ethyl groups on native cysteines to map the structural transitions of the sarcoplasmic reticulum Ca(2+)-ATPase, a 110 kDa transmembrane enzyme that transports Ca(2+) into the sarcoplasmic reticulum. Using magic angle spinning NMR, we monitored the chemical shifts of the methylene and methyl groups of the derivatized cysteine residues along the major steps of the enzymatic cycle. The methylene chemical shifts are sensitive to the ATPase conformational changes induced upon nucleotide and Ca(2+) ion binding and are ideal probes for active and inactive states of the enzyme. This new approach is extendable to large mammalian enzymes and signaling proteins with native or engineered cysteine residues in their amino acid sequence. PMID:26650884

  1. (13)C solid-state NMR analysis of the most common pharmaceutical excipients used in solid drug formulations, Part I: Chemical shifts assignment.

    PubMed

    Pisklak, Dariusz Maciej; Zielińska-Pisklak, Monika Agnieszka; Szeleszczuk, Łukasz; Wawer, Iwona

    2016-04-15

    Solid-state NMR is an excellent and useful method for analyzing solid-state forms of drugs. In the (13)C CP/MAS NMR spectra of the solid dosage forms many of the signals originate from the excipients and should be distinguished from those of active pharmaceutical ingredient (API). In this work the most common pharmaceutical excipients used in the solid drug formulations: anhydrous α-lactose, α-lactose monohydrate, mannitol, sucrose, sorbitol, sodium starch glycolate type A and B, starch of different origin, microcrystalline cellulose, hypromellose, ethylcellulose, methylcellulose, hydroxyethylcellulose, sodium alginate, magnesium stearate, sodium laurilsulfate and Kollidon(®) were analyzed. Their (13)C CP/MAS NMR spectra were recorded and the signals were assigned, employing the results (R(2): 0.948-0.998) of GIPAW calculations and theoretical chemical shifts. The (13)C ssNMR spectra for some of the studied excipients have not been published before while for the other signals in the spectra they were not properly assigned or the assignments were not correct. The results summarize and complement the data on the (13)C ssNMR analysis of the most common pharmaceutical excipients and are essential for further NMR studies of API-excipient interactions in the pharmaceutical formulations. PMID:26845204

  2. High-yield expression and purification of isotopically labeled cytochrome P450 monooxygenases for solid-state NMR spectroscopy

    PubMed Central

    Rupasinghe, Sanjeewa G.; Duan, Hui; Frericks Schmidt, Heather L.; Berthold, Deborah A.; Rienstra, Chad M.; Schuler, Mary A.

    2008-01-01

    Cytochrome P450 monooxygenases (P450s), which represent the major group of drug metabolizing enzymes in humans, also catalyze important synthetic and detoxicative reactions in insects, plants and many microbes. Flexibilities in their catalytic sites and membrane associations are thought to play central roles in substrate binding and catalytic specificity. To date, E. coli expression strategies for structural analysis of eukaryotic membrane-bound P450s by X-ray crystallography have necessitated full or partial removal of their N-terminal signal anchor domain (SAD) and, often, replacement of residues more peripherally associated with the membrane (such as the F-G loop region). Even with these modifications, investigations of P450 structural flexibility remain challenging with multiple single crystal conditions needed to identify spatial variations between substrate-free and different substrate-bound forms. To overcome these limitations, we have developed methods for the efficient expression of 13C- and 15N-labeled P450s and analysis of their structures by magic-angle spinning solid-state NMR (SSNMR) spectroscopy. In the presence of co-expressed GroEL and GroES chaperones, full-length (53 kDa) Arabidopsis 13C,15N-labeled CYP98A3 is expressed at yields of 2–4 mg per liter of minimal media without the necessity of generating side chain modifications or N-terminal deletions. Precipitated CYP98A3 generates high quality SSNMR spectra consistent with a homogeneous, folded protein. These data highlight the potential of these methodologies to contribute to the structural analysis of membrane-bound proteins. PMID:18005930

  3. Active-site structure of the thermophilic Foc-subunit ring in membranes elucidated by solid-state NMR.

    PubMed

    Kang, Su-Jin; Todokoro, Yasuto; Yumen, Ikuko; Shen, Bo; Iwasaki, Iku; Suzuki, Toshiharu; Miyagi, Atsushi; Yoshida, Masasuke; Fujiwara, Toshimichi; Akutsu, Hideo

    2014-01-21

    FoF1-ATP synthase uses the electrochemical potential across membranes or ATP hydrolysis to rotate the Foc-subunit ring. To elucidate the underlying mechanism, we carried out a structural analysis focused on the active site of the thermophilic c-subunit (TFoc) ring in membranes with a solid-state NMR method developed for this purpose. We used stereo-array isotope labeling (SAIL) with a cell-free system to highlight the target. TFoc oligomers were purified using a virtual ring His tag. The membrane-reconstituted TFoc oligomer was confirmed to be a ring indistinguishable from that expressed in E.coli on the basis of the H(+)-translocation activity and high-speed atomic force microscopic images. For the analysis of the active site, 2D (13)C-(13)C correlation spectra of TFoc rings labeled with SAIL-Glu and -Asn were recorded. Complete signal assignment could be performed with the aid of the C(?)i+1-C(?)i correlation spectrum of specifically (13)C,(15)N-labeled TFoc rings. The C(?) chemical shift of Glu-56, which is essential for H(+) translocation, and related crosspeaks revealed that its carboxyl group is protonated in the membrane, forming the H(+)-locked conformation with Asn-23. The chemical shift of Asp-61 C(?) of the E.coli c ring indicated an involvement of a water molecule in the H(+) locking, in contrast to the involvement of Asn-23 in the TFoc ring, suggesting two different means of proton storage in the c rings. PMID:24461014

  4. Polydisperse methyl β-cyclodextrin-epichlorohydrin polymers: variable contact time (13)C CP-MAS solid-state NMR characterization.

    PubMed

    Mallard, Isabelle; Baudelet, Davy; Castiglione, Franca; Ferro, Monica; Panzeri, Walter; Ragg, Enzio; Mele, Andrea

    2015-01-01

    The polymerization of partially methylated β-cyclodextrin (CRYSMEB) with epichlorohydrin was carried out in the presence of a known amount of toluene as imprinting agent. Three different preparations (D1, D2 and D3) of imprinted polymers were obtained and characterized by solid-state (13)C NMR spectroscopy under cross-polarization magic angle spinning (CP-MAS) conditions. The polymers were prepared by using the same synthetic conditions but with different molar ratios of imprinting agent/monomer, leading to morphologically equivalent materials but with different absorption properties. The main purpose of the work was to find a suitable spectroscopic descriptor accounting for the different imprinting process in three homogeneous polymeric networks. The polymers were characterized by studying the kinetics of the cross-polarization process. This approach is based on variable contact time CP-MAS spectra, referred to as VCP-MAS. The analysis of the VCP-MAS spectra provided two relaxation parameters: T CH (the CP time constant) and T 1ρ (the proton spin-lattice relaxation time in the rotating frame). The results and the analysis presented in the paper pointed out that T CH is sensitive to the imprinting process, showing variations related to the toluene/cyclodextrin molar ratio used for the preparation of the materials. Conversely, the observed values of T 1ρ did not show dramatic variations with the imprinting protocol, but rather confirmed that the three polymers are morphologically similar. Thus the combined use of T CH and T 1ρ can be helpful for the characterization and fine tuning of imprinted polymeric matrices. PMID:26877800

  5. Magic-angle spinning solid-state NMR spectroscopy of nanodisc-embedded human CYP3A4.

    PubMed

    Kijac, Aleksandra Z; Li, Ying; Sligar, Stephen G; Rienstra, Chad M

    2007-12-01

    Cytochrome P450 (CYP) 3A4 contributes to the metabolism of approximately 50% of commercial drugs by oxidizing a large number of structurally diverse substrates. Like other endoplasmic reticulum-localized P450s, CYP3A4 contains a membrane-anchoring N-terminal helix and a significant number of hydrophobic domains, important for the interaction between CYP3A4 and the membrane. Although the membrane affects specificity of CYP3A4 ligand binding, the structural details of the interaction have not been revealed so far because X-ray crystallography studies are available only for the soluble domain of CYP3A4. Here we report sample preparation and initial magic-angle spinning (MAS) solid-state NMR (SSNMR) of CYP3A4 (Delta3-12) embedded in a nanoscale membrane bilayer, or Nanodisc. The growth protocol yields approximately 2.5 mg of the enzymatically active, uniformly 13C,15N-enriched CYP3A4 from 1 L of growth medium. Polyethylene glycol 3350-precipitated CYP3A4 in Nanodiscs yields spectra of high resolution and sensitivity, consistent with a folded, homogeneous protein. CYP3A4 in Nanodiscs remains enzymatically active throughout the precipitation protocol as monitored by bromocriptine binding. The 13C line widths measured from 13C-13C 2D chemical shift correlation spectra are approximately 0.5 ppm. The secondary structure distribution within several amino acid types determined from 13C chemical shifts is consistent with the ligand-free X-ray structures. These results demonstrate that MAS SSNMR can be performed on Nanodisc-embedded membrane proteins in a folded, active state. The combination of SSNMR and Nanodisc methodologies opens up new possibilities for obtaining structural information on CYP3A4 and other integral membrane proteins with full retention of functionality. PMID:17985934

  6. Synthesis, characterization, and solid-state NMR investigation of organically modified bentonites and their composites with LDPE.

    PubMed

    Borsacchi, Silvia; Sudhakaran, Umayal; Geppi, Marco; Ricci, Lucia; Liuzzo, Vincenzo; Ruggeri, Giacomo

    2013-07-23

    Polymer/clay nanocomposites show remarkably improved properties (mechanical properties, as well as decreased gas permeability and flammability, etc.) with respect to their microscale counterparts and pristine polymers. Due to the substantially apolar character of most of the organic polymers, natural occurring hydrophilic clays are modified into organophilic clays with consequent increase of the polymer/clay compatibility. Different strategies have been developed for the preparation of nanocomposites with improved properties, especially aimed at achieving the best dispersion of clay platelets in the polymer matrix. In this paper we present the preparation and characterization of polymer/clay nanocomposites composed of low-density polyethylene (LDPE) and natural clay, montmorillonite-containing bentonite. Two different forms of the clay have been considered: the first, a commercial organophilic bentonite (Nanofil 15), obtained by exchanging the natural cations with dimethyldioctadecylammonium (2C18) cations, and the second, obtained by performing a grafting reaction of an alkoxysilane containing a polymerizable group, 3-(trimethoxysilyl)propyl methacrylate (TSPM), onto Nanofil 15. Both the clays and LDPE/clay nanocomposites were characterized by thermal, FT-IR, and X-ray diffraction techniques. The samples were also investigated by means of (29)Si, (13)C, and (1)H solid-state NMR, obtaining information on the structural properties of the modified clays. Moreover, by exploiting the effect of bentonite paramagnetic (Fe(3+)) ions on proton spin-lattice relaxation times (T1's), useful information about the extent of the polymer-clay dispersion and their interfacial interactions could be obtained. PMID:23786424

  7. Solid-state NMR spectroscopic study of chromophore-protein interactions in the Pr ground state of plant phytochrome A.

    PubMed

    Song, Chen; Essen, Lars-Oliver; Gärtner, Wolfgang; Hughes, Jon; Matysik, Jörg

    2012-05-01

    Despite extensive study, the molecular structure of the chromophore-binding pocket of phytochrome A (phyA), the principal photoreceptor controlling photomorphogenesis in plants, has not yet been successfully resolved. Here, we report a series of two-dimensional (2-D) magic-angle spinning solid-state NMR experiments on the recombinant N-terminal, 65-kDa PAS-GAF-PHY light-sensing module of phytochrome A3 from oat (Avena sativa), assembled with uniformly 13C- and 15N-labeled phycocyanobilin (u-[13C,15N]-PCB-As.phyA3). The Pr state of this protein was studied regarding the electronic structure of the chromophore and its interactions with the proximal amino acids. Using 2-D 13C-13C and 1H-15N experiments, a complete set of 13C and 15N assignments for the chromophore were obtained. Also, a large number of 1H-13C distance restraints between the chromophore and its binding pocket were revealed by interfacial heteronuclear correlation spectroscopy. 13C doublings of the chromophore A-ring region and the C-ring carboxylate moiety, together with the observation of two Pr isoforms, Pr-I and Pr-II, demonstrate the local mobility of the chromophore and the plasticity of its protein environment. It appears that the interactions and dynamics in the binding pocket of phyA in the Pr state are remarkably similar to those of cyanobacterial phytochrome (Cph1). The N-terminus of the region modeled (residues 56-66 of phyA) is highly mobile. Differences in the regulatory processes involved in plant and Cph1 phytochromes are discussed. PMID:22419823

  8. Polydisperse methyl β-cyclodextrin–epichlorohydrin polymers: variable contact time 13C CP-MAS solid-state NMR characterization

    PubMed Central

    Mallard, Isabelle; Baudelet, Davy; Castiglione, Franca; Ferro, Monica; Panzeri, Walter; Ragg, Enzio

    2015-01-01

    Summary The polymerization of partially methylated β-cyclodextrin (CRYSMEB) with epichlorohydrin was carried out in the presence of a known amount of toluene as imprinting agent. Three different preparations (D1, D2 and D3) of imprinted polymers were obtained and characterized by solid-state 13C NMR spectroscopy under cross-polarization magic angle spinning (CP-MAS) conditions. The polymers were prepared by using the same synthetic conditions but with different molar ratios of imprinting agent/monomer, leading to morphologically equivalent materials but with different absorption properties. The main purpose of the work was to find a suitable spectroscopic descriptor accounting for the different imprinting process in three homogeneous polymeric networks. The polymers were characterized by studying the kinetics of the cross-polarization process. This approach is based on variable contact time CP-MAS spectra, referred to as VCP-MAS. The analysis of the VCP-MAS spectra provided two relaxation parameters: T CH (the CP time constant) and T 1ρ (the proton spin-lattice relaxation time in the rotating frame). The results and the analysis presented in the paper pointed out that T CH is sensitive to the imprinting process, showing variations related to the toluene/cyclodextrin molar ratio used for the preparation of the materials. Conversely, the observed values of T 1ρ did not show dramatic variations with the imprinting protocol, but rather confirmed that the three polymers are morphologically similar. Thus the combined use of T CH and T 1ρ can be helpful for the characterization and fine tuning of imprinted polymeric matrices. PMID:26877800

  9. Direct Speciation of Phosphorus in Alum-Amended Poultry Litter: Solid-State 31P NMR Investigation

    SciTech Connect

    Hunger, Stefan; Cho, Herman M.; Sims, James T.; Sparks, Donald L.

    2004-02-01

    Amending poultry litter (PL) with aluminum sulfate (alum) has proven to be effective in reducing water-soluble phosphorus (P) in the litter and in runoff from fields that have received PL applications; it has therefore been suggested as a best management practice. Although its effectiveness has been demonstrated on a macroscopic scale in the field, little is known about P speciation in either alumamended or unamended litter. This knowledge is important for the evaluation of the long-term stability and bioavailability of P, which is a necessary prerequisite for the assessment of the sustainability of intensive poultry operations. Both solid state MAS and CP-MAS {sup 31}P NMR as well as {sup 31}P({sup 27}Al) TRAPDOR were used to investigate P speciation in alumamended and unamended PL. The results indicate the presence of a complex mixture of organic and inorganic orthophosphate phases. A calcium phosphate phase, probably a surface precipitate on calcium carbonate, could be identified in both unamended and alum-amended PL, as well as physically bound HPO{sub 4}{sup 2-}. Phosphate associated with Al was found in the alum-amended PL, most probably a mixture of a poorly ordered wavellite and phosphate surface complexes on aluminum hydroxide that had been formed by the hydrolysis of alum. However, a complex mixture of organic and inorganic phosphate species could not be resolved. Phosphate associated with Al comprised on average 40{+-}14% of the total P in alum-amended PL, whereas calcium phosphate phases comprised on average 7{+-}4% in the alum-amended PL and 14{+-}5% in the unamended PL.

  10. Solid-state NMR Reveals the Carbon-based Molecular Architecture of Cryptococcus neoformans Fungal Eumelanins in the Cell Wall.

    PubMed

    Chatterjee, Subhasish; Prados-Rosales, Rafael; Itin, Boris; Casadevall, Arturo; Stark, Ruth E

    2015-05-29

    Melanin pigments protect against both ionizing radiation and free radicals and have potential soil remediation capabilities. Eumelanins produced by pathogenic Cryptococcus neoformans fungi are virulence factors that render the fungal cells resistant to host defenses and certain antifungal drugs. Because of their insoluble and amorphous characteristics, neither the pigment bonding framework nor the cellular interactions underlying melanization of C. neoformans have yielded to comprehensive molecular-scale investigation. This study used the C. neoformans requirement of exogenous obligatory catecholamine precursors for melanization to produce isotopically enriched pigment "ghosts" and applied 2D (13)C-(13)C correlation solid-state NMR to reveal the carbon-based architecture of intact natural eumelanin assemblies in fungal cells. We demonstrated that the aliphatic moieties of solid C. neoformans melanin ghosts include cell-wall components derived from polysaccharides and/or chitin that are associated proximally with lipid membrane constituents. Prior to development of the mature aromatic fungal pigment, these aliphatic moieties form a chemically resistant framework that could serve as the scaffold for melanin synthesis. The indole-based core aromatic moieties show interconnections that are consistent with proposed melanin structures consisting of stacked planar assemblies, which are associated spatially with the aliphatic scaffold. The pyrrole aromatic carbons of the pigments bind covalently to the aliphatic framework via glycoside or glyceride functional groups. These findings establish that the structure of the pigment assembly changes with time and provide the first biophysical information on the mechanism by which melanin is assembled in the fungal cell wall, offering vital insights that can advance the design of bioinspired conductive nanomaterials and novel therapeutics. PMID:25825492

  11. Active-Site Structure of the Thermophilic Foc-Subunit Ring in Membranes Elucidated by Solid-State NMR

    PubMed Central

    Kang, Su-Jin; Todokoro, Yasuto; Yumen, Ikuko; Shen, Bo; Iwasaki, Iku; Suzuki, Toshiharu; Miyagi, Atsushi; Yoshida, Masasuke; Fujiwara, Toshimichi; Akutsu, Hideo

    2014-01-01

    FoF1-ATP synthase uses the electrochemical potential across membranes or ATP hydrolysis to rotate the Foc-subunit ring. To elucidate the underlying mechanism, we carried out a structural analysis focused on the active site of the thermophilic c-subunit (TFoc) ring in membranes with a solid-state NMR method developed for this purpose. We used stereo-array isotope labeling (SAIL) with a cell-free system to highlight the target. TFoc oligomers were purified using a virtual ring His tag. The membrane-reconstituted TFoc oligomer was confirmed to be a ring indistinguishable from that expressed in E. coli on the basis of the H+-translocation activity and high-speed atomic force microscopic images. For the analysis of the active site, 2D 13C-13C correlation spectra of TFoc rings labeled with SAIL-Glu and -Asn were recorded. Complete signal assignment could be performed with the aid of the Cαi+1-Cαi correlation spectrum of specifically 13C,15N-labeled TFoc rings. The Cδ chemical shift of Glu-56, which is essential for H+ translocation, and related crosspeaks revealed that its carboxyl group is protonated in the membrane, forming the H+-locked conformation with Asn-23. The chemical shift of Asp-61 Cγ of the E. coli c ring indicated an involvement of a water molecule in the H+ locking, in contrast to the involvement of Asn-23 in the TFoc ring, suggesting two different means of proton storage in the c rings. PMID:24461014

  12. Mechanism of formation of humus coatings on mineral surfaces 3. Composition of adsorbed organic acids from compost leachate on alumina by solid-state 13C NMR

    USGS Publications Warehouse

    Wershaw, R. L.; Llaguno, E.C.; Leenheer, J.A.

    1996-01-01

    The adsorption of compost leachate DOC on alumina is used as a model for elucidation of the mechanism of formation of natural organic coatings on hydrous metal oxide surfaces in soils and sediments. Compost leachate DOC is composed mainly of organic acid molecules. The solid-state 13C NMR spectra of these organic acids indicate that they are very similar in composition to aquatic humic substances. Changes in the solid-state 13C NMR spectra of compost leachate DOC fractions adsorbed on alumina indicate that the DOC molecules are most likely adsorbed on metal oxide surfaces through a combination of polar and hydrophobic interaction mechanisms. This combination of polar and hydrophobic mechanism leads to the formation of bilayer coatings of the leachate molecules on the oxide surfaces.

  13. Amyloid Hydrogen Bonding Polymorphism Evaluated by (15)N{(17)O}REAPDOR Solid-State NMR and Ultra-High Resolution Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.

    PubMed

    Wei, Juan; Antzutkin, Oleg N; Filippov, Andrei V; Iuga, Dinu; Lam, Pui Yiu; Barrow, Mark P; Dupree, Ray; Brown, Steven P; O'Connor, Peter B

    2016-04-12

    A combined approach, using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) and solid-state NMR (Nuclear Magnetic Resonance), shows a high degree of polymorphism exhibited by Aβ species in forming hydrogen-bonded networks. Two Alzheimer's Aβ peptides, Ac-Aβ16-22-NH2 and Aβ11-25, selectively labeled with (17)O and (15)N at specific amino acid residues were investigated. The total amount of peptides labeled with (17)O as measured by FTICR-MS enabled the interpretation of dephasing observed in (15)N{(17)O}REAPDOR solid-state NMR experiments. Specifically, about one-third of the Aβ peptides were found to be involved in the formation of a specific >C═(17)O···H-(15)N hydrogen bond with their neighbor peptide molecules, and we hypothesize that the rest of the molecules undergo ± n off-registry shifts in their hydrogen bonding networks. PMID:26983928

  14. High-resolution paramagnetically enhanced solid-state NMR spectroscopy of membrane proteins at fast magic angle spinning.

    PubMed

    Ward, Meaghan E; Wang, Shenlin; Krishnamurthy, Sridevi; Hutchins, Howard; Fey, Michael; Brown, Leonid S; Ladizhansky, Vladimir

    2014-01-01

    Magic angle spinning nuclear magnetic resonance (MAS NMR) is well suited for the study of membrane proteins in membrane mimetic and native membrane environments. These experiments often suffer from low sensitivity, due in part to the long recycle delays required for magnetization and probe recovery, as well as detection of low gamma nuclei. In ultrafast MAS experiments sensitivity can be enhanced through the use of low power sequences combined with paramagnetically enhanced relaxation times to reduce recycle delays, as well as proton detected experiments. In this work we investigate the sensitivity of (13)C and (1)H detected experiments applied to 27 kDa membrane proteins reconstituted in lipids and packed in small 1.3 mm MAS NMR rotors. We demonstrate that spin diffusion is sufficient to uniformly distribute paramagnetic relaxation enhancement provided by either covalently bound or dissolved CuEDTA over 7TM alpha helical membrane proteins. Using paramagnetic enhancement and low power decoupling in carbon detected experiments we can recycle experiments ~13 times faster than under traditional conditions. However, due to the small sample volume the overall sensitivity per unit time is still lower than that seen in the 3.2 mm probe. Proton detected experiments, however, showed increased efficiency and it was found that the 1.3 mm probe could achieve sensitivity comparable to that of the 3.2 mm in a given amount of time. This is an attractive prospect for samples of limited quantity, as this allows for a reduction in the amount of protein that needs to be produced without the necessity for increased experimental time. PMID:24338448

  15. Direct observation of DME carbonylation in the different channels of H-MOR zeolite by continuous-flow solid-state NMR spectroscopy.

    PubMed

    He, Ting; Ren, Pengju; Liu, Xianchun; Xu, Shutao; Han, Xiuwen; Bao, Xinhe

    2015-12-01

    The dynamic evolution of acetyl intermediates in the two different channels of H-mordenite (H-MOR) zeolite during dimethyl ether (DME) carbonylation is tracked by using in situ solid-state NMR spectroscopy under continuous-flow conditions. Thus, the reaction path via methyl acetate produced over active sites in 8 member ring (MR) channels, followed by diffusion into 12 MR channels, is proposed. PMID:26451500

  16. Non-destructive detection of methionine sulfoxide in the resilium of a surf clam by solid-state 13C-NMR spectroscopy.

    PubMed

    Kikuchi, Y; Tamiya, N; Nozawa, T; Hatano, M

    1982-07-01

    Methionine sulfoxide was detected in the resilium (internal hinge ligament) of a surf clam by high-resolution solid-state 13C-NMR spectroscopy involving cross-polarization and magic angle spinning, using no chemical procedure. The results support the previous report [Kikuchi, Y. and Tamiya, N. (1981) J. Biochem. (Tokyo) 89, 1975-1976] on a high content of methionine sulfoxide observed by chemical methods in the resilium protein of surf clam species. PMID:7117255

  17. Local environment and composition of magnesium gallium layered double hydroxides determined from solid-state 1H and 71Ga NMR spectroscopy

    SciTech Connect

    Petersen, Line B.; Lipton, Andrew S.; Zorin, Vadim; Nielsen, Ulla Gro

    2014-11-01

    Ordering of gallium(III) in a series of magnesium gallium layered double hydroxides (LDH’s), [Mg1-xGax(OH)2(NO3)x yH2O], was determined using solid-state 1H and 71Ga NMR spectroscopy. Depletion of Ga in these LDH’s is demonstrated to be the result of soluble [Ga(OH)4]-complexes formed during synthesis.

  18. Preservation of proteinaceous material during the degradation of the green alga Botryococcus braunii: A solid-state 2D 15N 13C NMR spectroscopy study

    NASA Astrophysics Data System (ADS)

    Zang, Xu; Nguyen, Reno T.; Harvey, H. Rodger; Knicker, Heike; Hatcher, Patrick G.

    2001-10-01

    Using solid-state cross-polarization-magic-angle-spinning (CPMAS) 13C and 15N nuclear magnetic resonance (NMR) and 2-D double cross polarization (DCP) MAS 15N 13C NMR techniques, microbially degraded Botryococcus braunii was analyzed to study the chemical nature of organic nitrogen in the algal residue. The amide linkage, as found in protein, was observed as the major nitrogen component in 201-day-old degraded algae. No significant amount of heterocyclic nitrogen, or evidence for melanoidin products, was found. The results strongly suggest that proteinaceous material can survive early diagenesis and be preserved via its encapsulation by refractory, macromolecular, organic matter.

  19. Structure determination of uniformly (13)C, (15)N labeled protein using qualitative distance restraints from MAS solid-state (13)C-NMR observed paramagnetic relaxation enhancement.

    PubMed

    Tamaki, Hajime; Egawa, Ayako; Kido, Kouki; Kameda, Tomoshi; Kamiya, Masakatsu; Kikukawa, Takashi; Aizawa, Tomoyasu; Fujiwara, Toshimichi; Demura, Makoto

    2016-01-01

    Magic angle spinning (MAS) solid-state nuclear magnetic resonance (NMR) is a powerful method for structure determination of insoluble biomolecules. However, structure determination by MAS solid-state NMR remains challenging because it is difficult to obtain a sufficient amount of distance restraints owing to spectral complexity. Collection of distance restraints from paramagnetic relaxation enhancement (PRE) is a promising approach to alleviate this barrier. However, the precision of distance restraints provided by PRE is limited in solid-state NMR because of incomplete averaged interactions and intermolecular PREs. In this report, the backbone structure of the B1 domain of streptococcal protein G (GB1) has been successfully determined by combining the CS-Rosetta protocol and qualitative PRE restraints. The derived structure has a Cα RMSD of 1.49 Å relative to the X-ray structure. It is noteworthy that our protocol can determine the correct structure from only three cysteine-EDTA-Mn(2+) mutants because this number of PRE sites is insufficient when using a conventional structure calculation method based on restrained molecular dynamics and simulated annealing. This study shows that qualitative PRE restraints can be employed effectively for protein structure determination from a limited conformational sampling space using a protein fragment library. PMID:26728076

  20. Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra

    NASA Astrophysics Data System (ADS)

    Gopinath, T.; Veglia, Gianluigi

    2013-05-01

    We propose a general method that enables the acquisition of multiple 2D and 3D solid-state NMR spectra for U-13C, 15N-labeled proteins. This method, called MEIOSIS (Multiple ExperIments via Orphan SpIn operatorS), makes it possible to detect four coherence transfer pathways simultaneously, utilizing orphan (i.e., neglected) spin operators of nuclear spin polarization generated during 15N-13C cross polarization (CP). In the MEIOSIS experiments, two phase-encoded free-induction decays are decoded into independent nuclear polarization pathways using Hadamard transformations. As a proof of principle, we show the acquisition of multiple 2D and 3D spectra of U-13C, 15N-labeled microcrystalline ubiquitin. Hadamard decoding of CP coherences into multiple independent spin operators is a new concept in solid-state NMR and is extendable to many other multidimensional experiments. The MEIOSIS method will increase the throughput of solid-state NMR techniques for microcrystalline proteins, membrane proteins, and protein fibrils.

  1. Orphan spin operators enable the acquisition of multiple 2D and 3D magic angle spinning solid-state NMR spectra

    PubMed Central

    Gopinath, T.; Veglia, Gianluigi

    2013-01-01

    We propose a general method that enables the acquisition of multiple 2D and 3D solid-state NMR spectra for U-13C, 15N-labeled proteins. This method, called MEIOSIS (Multiple ExperIments via Orphan SpIn operatorS), makes it possible to detect four coherence transfer pathways simultaneously, utilizing orphan (i.e., neglected) spin operators of nuclear spin polarization generated during 15N-13C cross polarization (CP). In the MEIOSIS experiments, two phase-encoded free-induction decays are decoded into independent nuclear polarization pathways using Hadamard transformations. As a proof of principle, we show the acquisition of multiple 2D and 3D spectra of U-13C, 15N-labeled microcrystalline ubiquitin. Hadamard decoding of CP coherences into multiple independent spin operators is a new concept in solid-state NMR and is extendable to many other multidimensional experiments. The MEIOSIS method will increase the throughput of solid-state NMR techniques for microcrystalline proteins, membrane proteins, and protein fibrils. PMID:23676036

  2. NMR-Based Structural Modeling of Graphite Oxide Using Multidimensional 13C Solid-State NMR and ab Initio Chemical Shift Calculations

    PubMed Central

    2010-01-01

    Chemically modified graphenes and other graphite-based materials have attracted growing interest for their unique potential as lightweight electronic and structural nanomaterials. It is an important challenge to construct structural models of noncrystalline graphite-based materials on the basis of NMR or other spectroscopic data. To address this challenge, a solid-state NMR (SSNMR)-based structural modeling approach is presented on graphite oxide (GO), which is a prominent precursor and interesting benchmark system of modified graphene. An experimental 2D 13C double-quantum/single-quantum correlation SSNMR spectrum of 13C-labeled GO was compared with spectra simulated for different structural models using ab initio geometry optimization and chemical shift calculations. The results show that the spectral features of the GO sample are best reproduced by a geometry-optimized structural model that is based on the Lerf−Klinowski model (Lerf, A. et al. Phys. Chem. B1998, 102, 4477); this model is composed of interconnected sp2, 1,2-epoxide, and COH carbons. This study also convincingly excludes the possibility of other previously proposed models, including the highly oxidized structures involving 1,3-epoxide carbons (Szabo, I. et al. Chem. Mater.2006, 18, 2740). 13C chemical shift anisotropy (CSA) patterns measured by a 2D 13C CSA/isotropic shift correlation SSNMR were well reproduced by the chemical shift tensor obtained by the ab initio calculation for the former model. The approach presented here is likely to be applicable to other chemically modified graphenes and graphite-based systems. PMID:20359218

  3. Characteristics and degradation of carbon and phosphorus from aquatic macrophytes in lakes: Insights from solid-state (13)C NMR and solution (31)P NMR spectroscopy.

    PubMed

    Liu, Shasha; Zhu, Yuanrong; Meng, Wei; He, Zhongqi; Feng, Weiying; Zhang, Chen; Giesy, John P

    2016-02-01

    Water extractable organic matter (WEOM) derived from macrophytes plays an important role in biogeochemical cycling of nutrients, including carbon (C), nitrogen (N) and phosphorus (P) in lakes. However, reports of their composition and degradation in natural waters are scarce. Therefore, compositions and degradation of WEOM derived from six aquatic macrophytes species of Tai Lake, China, were investigated by use of solid-state (13)C NMR and solution (31)P NMR spectroscopy. Carbohydrates were the predominant constituents of WEOM fractions, followed by carboxylic acid. Orthophosphate (ortho-P) was the dominant form of P (78.7% of total dissolved P) in the water extracts, followed by monoester P (mono-P) (20.6%) and little diester P (0.65%). The proportion of mono-P in total P species increased with the percentage of O-alkyl and O-C-O increasing in the WEOM, which is likely due to degradation and dissolution of biological membranes and RNA from aquatic plants. Whereas the proportion of mono-P decreased with alkyl-C, NCH/OCH3 and COO/N-C=O increasing, which may be owing to the insoluble compounds including C functional groups of alkyl-C, NCH/OCH3 and COO/N-C=O, such as aliphatic biopolymers, lignin and peptides. Based on the results of this study and information in the literature about water column and sediment, we propose that WEOM, dominated by polysaccharides, are the most labile and bioavailable component in debris of macrophytes. Additionally, these WEOMs would also be a potential source for bioavailable organic P (e.g., RNA, DNA and phytate) for lakes. PMID:26624522

  4. Spin dynamics in the modulation frame: application to homonuclear recoupling in magic angle spinning solid-state NMR.

    PubMed

    De Paëpe, Gaël; Lewandowski, Józef R; Griffin, Robert G

    2008-03-28

    We introduce a family of solid-state NMR pulse sequences that generalizes the concept of second averaging in the modulation frame and therefore provides a new approach to perform magic angle spinning dipolar recoupling experiments. Here, we focus on two particular recoupling mechanisms-cosine modulated rotary resonance (CMpRR) and cosine modulated recoupling with isotropic chemical shift reintroduction (COMICS). The first technique, CMpRR, is based on a cosine modulation of the rf phase and yields broadband double-quantum (DQ) (13)C recoupling using >70 kHz omega(1,C)/2pi rf field for the spinning frequency omega(r)/2=10-30 kHz and (1)H Larmor frequency omega(0,H)/2pi up to 900 MHz. Importantly, for p>or=5, CMpRR recouples efficiently in the absence of (1)H decoupling. Extension to lower p values (3.5

  5. The Stoichiometry of Synthetic Alunite as a Function of Hydrothermal Aging Investigated by Solid-State NMR Spectroscopy, Powder X-ray Diffraction and Infrared Spectroscopy

    SciTech Connect

    Grube, Elisabeth; Nielsen, Ulla Gro

    2015-05-01

    The stoichiometry of a series of synthetic alunite [nominally KAl3(SO4)2(OH)6] samples prepared by hydrothermal methods as a function of reaction time (1–31 days) has been investigated by powder X-ray diffraction, Fourier transform infrared spectroscopy as well as solid-state 1H and 27Al magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. The 1H MAS NMR spectra recorded at high magnetic field (21.1 T, 900 MHz) allowed for a clear separation of the different proton environments and for quantitative determination of the aluminum vacancy concentration as a function of time. The concentration of structural defects determined from, i.e., aluminum vacancies was reduced from 4 to 1 %, as the reaction time was extended from one to 31 days based on 1H MAS NMR. This was further supported by an increase of the unit cell parameter c, which is indicative of the relative concentration of potassium defects present, from 17.261(1) to 17.324(5) Å. Solid-state 27Al MAS NMR revealed a decrease in the defect concentration as a function of time and showed the presence of 7–10 % impurities in the samples.

  6. Identification of lithium-sulfur battery discharge products through 6Li and 33S solid-state MAS and 7Li solution NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Huff, Laura A.; Rapp, Jennifer L.; Baughman, Jessi A.; Rinaldi, Peter L.; Gewirth, Andrew A.

    2015-01-01

    6Li and 33S solid-state magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy was used to identify the discharge products in lithium-sulfur (Li-S) battery cathodes. Cathodes were stopped at different potentials throughout battery discharge and measured ex-situ to obtain chemical shifts and T2 relaxation rates of the products formed. The chemical shifts in the spectra of both 6Li and 33S NMR demonstrate that long-chain, soluble lithium polysulfide species formed at the beginning of discharge are indistinguishable from each other (similar chemical shifts), while short-chain, insoluble polysulfide species that form at the end of discharge (presumably Li2S2 and Li2S) have a different chemical shift, thus distinguishing them from the soluble long-chain products. T2 relaxation measurements of discharged cathodes were also performed which resulted in two groupings of T2 rates that follow a trend and support the previous conclusions that long-chain polysulfide species are converted to shorter chain species during discharge. Through the complementary techniques of 1-D 6Li and 33S solid-state MAS NMR spectroscopy, solution 7Li and 1H NMR spectroscopy, and T2 relaxation rate measurements, structural information about the discharge products of Li-S batteries is obtained.

  7. The stoichiometry of synthetic alunite as a function of hydrothermal aging investigated by solid-state NMR spectroscopy, powder X-ray diffraction and infrared spectroscopy

    NASA Astrophysics Data System (ADS)

    Grube, Elisabeth; Nielsen, Ulla Gro

    2015-05-01

    The stoichiometry of a series of synthetic alunite [nominally KAl3(SO4)2(OH)6] samples prepared by hydrothermal methods as a function of reaction time (1-31 days) has been investigated by powder X-ray diffraction, Fourier transform infrared spectroscopy as well as solid-state 1H and 27Al magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. The 1H MAS NMR spectra recorded at high magnetic field (21.1 T, 900 MHz) allowed for a clear separation of the different proton environments and for quantitative determination of the aluminum vacancy concentration as a function of time. The concentration of structural defects determined from, i.e., aluminum vacancies was reduced from 4 to 1 %, as the reaction time was extended from one to 31 days based on 1H MAS NMR. This was further supported by an increase of the unit cell parameter c, which is indicative of the relative concentration of potassium defects present, from 17.261(1) to 17.324(5) Å. Solid-state 27Al MAS NMR revealed a decrease in the defect concentration as a function of time and showed the presence of 7-10 % impurities in the samples.

  8. Structure and dynamics of poly(ethylene- co-1,5-hexadiene) as studied by solid state 13C NMR and quantum chemical calculations

    NASA Astrophysics Data System (ADS)

    Kurosu, Hiromichi; Yamamoto, Yuuri; Fujikawa, Aki; Kawabata, Emika; Sone, Masato; Naga, Naofumi

    2009-03-01

    Poly(ethylene- co-1,5-hexadiene) with 1,5-hexadiene (HD) contents of 1.8, 8.1, 9.7 and 20.3% was prepared by copolymerization of ethylene and HD involving intermolecular cyclization. Higher-order structures and dynamics of these samples were studied by solid state NMR and quantum chemistry. The 13C solid state NMR spectra and 13C spin-lattice relaxation time ( T1) of the samples were measured. The observed 13C CP/MAS and PST/MAS NMR spectra for all samples were decomposed into six peaks. The cyclopentane units incorporated in the main chain of polyethylene affected not only the crystalline structure but also the noncrystalline structure. This causes a trans-rich conformation in the noncrystalline region. Even in the melt-quenched samples, incorporation of the cyclopentane structure into the polyethylene chain suppresses the increase in the gauche structure in the noncrystalline region. Based on the 13C chemical shift of the methylene carbon, the low cyclopentane content sample assumes an orthorhombic crystal structure, and the high cyclopentane content samples assume a hexagonal-type chain packing. 13C spin-lattice relaxation times show that the crystalline region of the low cyclopentane content sample has two regions with different mobility, although the high cyclopentane content samples have only one region with a high mobility for each peak. Furthermore, quantum chemical calculations for the 13C NMR shieldings were carried out for precise assignment of the peaks.

  9. In situ solid state B-11 MAS-NMR studies of the thermal decomposition of ammonia borane: mechanistic studies of the hydrogen release pathways from a solid state hydrogen storage material

    SciTech Connect

    Stowe, Ashley C.; Shaw, Wendy J.; Linehan, John C.; Schmid, Benjamin; Autrey, Thomas

    2007-02-20

    The mechanism of hydrogen release from solid state ammonia borane (AB) has been investigated via in situ solid state 11B{1H} MAS-NMR techniques in external fields of 7.06 T and 18.8 T at a decomposition temperature of 88 oC, well below the reported melting point. The decomposition of AB is well described by an induction, nucleation and growth mechanistic pathway. During the induction period, little hydrogen is released from AB; however, a new species identified as a mobile phase of AB is observed in the 11B NMR spectra. Subsequent to induction, at reaction times when hydrogen is initially being released, three additional species are observed: the diammoniate of diborane (DADB), [(NH3)2BH2]+[BH4]-, and two BH2N2 species believed to be the linear (NH3BH2NH2BH3) and cyclic dimer (NH2BH2)2 of aminoborane. At longer reaction times the sharper features are replaced by broad, structureless peaks of a complex polymeric aminoborane (PAB) containing both BH2N2 and BHN3 species. We propose the following mechanistic model for the induction, nucleation and growth for AB decomposition leading to formation of hydrogen: (1) an induction period that yields a mobile phase of AB caused by disruption of the dihydrogen bonds, (2) nucleation that yields reactive DADB from the mobile AB and (3) growth that includes a bimolecular reaction between DADB and AB to release the stored hydrogen. Support for this work by the U.S. Department of Energy, Office of Science, Basic Energy Sciences is gratefully acknowledged. A portion of the research described in this paper was performed in the Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the U.S. Department of Energy's Office of Biological and Environmental Research and located at Pacific Northwest National Laboratory.

  10. Solid-state NMR studies of the membrane-bound closed state of the colicin E1 channel domain in lipid bilayers.

    PubMed Central

    Kim, Y.; Valentine, K.; Opella, S. J.; Schendel, S. L.; Cramer, W. A.

    1998-01-01

    The colicin E1 channel polypeptide was shown to be organized anisotropically in membranes by solid-state NMR analysis of samples of uniformly 15N-labeled protein in oriented planar phospholipid bilayers. The 190 residue C-terminal colicin E1 channel domain is the largest polypeptide to have been characterized by 15N solid-state NMR spectroscopy in oriented membrane bilayers. The 15N-NMR spectra of the colicin E1 show that: (1) the structure and dynamics are independent of anionic lipid content in both oriented and unoriented samples; (2) assuming the secondary structure of the polypeptide is helical, there are both trans-membrane and in-plane helical segments; (3) trans-membrane helices account for approximately 20-25% of the channel polypeptide, which is equivalent to 38-48 residues of the 190-residue polypeptide. The results of the two-dimensional PISEMA spectrum are interpreted in terms of a single trans-membrane helical hairpin inserted into the bilayer from each channel molecule. These data are also consistent with this helical hairpin being derived from the 38-residue hydrophobic segment near the C-terminus of the colicin E1 channel polypeptide. PMID:9521110

  11. A Monte Carlo/Simulated Annealing Algorithm for Sequential Resonance Assignment in Solid State NMR of Uniformly Labeled Proteins with Magic-Angle Spinning

    PubMed Central

    Tycko, Robert; Hu, Kan-Nian

    2010-01-01

    We describe a computational approach to sequential resonance assignment in solid state NMR studies of uniformly 15N,13C-labeled proteins with magic-angle spinning. As input, the algorithm uses only the protein sequence and lists of 15N/13Cα crosspeaks from 2D NCACX and NCOCX spectra that include possible residue-type assignments of each crosspeak. Assignment of crosspeaks to specific residues is carried out by a Monte Carlo/simulated annealing algorithm, implemented in the program MC_ASSIGN1. The algorithm tolerates substantial ambiguity in residue-type assignments and coexistence of visible and invisible segments in the protein sequence. We use MC_ASSIGN1 and our own 2D spectra to replicate and extend the sequential assignments for uniformly labeled HET-s(218-289) fibrils previously determined manually by Siemer et al. (J. Biomolec. NMR, vol. 34, pp. 75-87, 2006) from a more extensive set of 2D and 3D spectra. Accurate assignments by MC_ASSIGN1 do not require data that are of exceptionally high quality. Use of MC_ASSIGN1 (and its extensions to other types of 2D and 3D data) is likely to alleviate many of the difficulties and uncertainties associated with manual resonance assignments in solid state NMR studies of uniformly labeled proteins, where spectral resolution and signal-to-noise are often sub-optimal. PMID:20547467

  12. Some studies on a solid state sulfur probe for coal gasification systems

    NASA Technical Reports Server (NTRS)

    Jacob, K. T.; Rao, D. B.; Nelson, H. G.

    1977-01-01

    Measurements on the solid electrolyte cell (Ar + H(2) + H(2)S/CaS + CaF(2) + (Pt)//CaF(2)//(Pt) + CaF(2) + CaS/H(2) + H(2)+Ar) show that the emf of the cell is directly related to the difference in sulfur potentials established at the Ar + H(2) + H(2)S/electrode interfaces. The electrodes convert the sulfur potential gradient across the calcium fluoride electrolyte into an equivalent fluorine potential gradient. Response time of the probe varies from approximately 9 hr at 990 K to 2.5 hr at 1225 K. The conversion of calcium sulfide and/or calcium fluoride into calcium oxide is not a problem anticipated in commercial coal gasification systems. Suggestions are presented for improving the cell for such commercial applications.

  13. A Technique for Increasing the Sensitivity of a Solid-State Fission Probe

    NASA Technical Reports Server (NTRS)

    Steinberg, Robert

    1961-01-01

    A small silicon p-n junction wafer, when coated with uranium 235, can be used as a compact fission probe for low power flux and power mapping. Because of the inverse relation between the magnitude of a neutron-induced fission pulse and the inherent capacitance of the detecting element (capacitance is proportional to area), the size, and hence the sensitivity, of the semiconductor detector has been limited. New developments in the field of semiconductor detectors have made it possible to fabricate large area detectors which are essentially free from the capacitance effect. However, preliminary results indicate that they are much more susceptible to radiation damage than the detectors described in this report and as such may not be suitable for flux mapping. increasing the sensitivity cannot be accomplished by simply fabricating a larger detector. It has been observed that by combining the silicon p-n junction wafers in a series configuration the capacitance effect can be bypassed, and a fission probe can be made with a resultant increase in sensitivity by a factor of ten while sustaining only a minor decrease in pulse height. Analysis further indicates that for n silicon wafers in series, if n(C(sub i)) + C(sub c)/C(sub b) less than 0.1 where C(sub i) and C(sub c) are the preamplifier input and cable capacitances, respectively, and C(sub b) is the junction capacitance of a single silicon wafer, there should be no substantial reduction in pulse height due to series circuitry.

  14. 33S NMR cryogenic probe for taurine detection.

    PubMed

    Hobo, Fumio; Takahashi, Masato; Maeda, Hideaki

    2009-03-01

    With the goal of a (33)S nuclear magnetic resonance (NMR) probe applicable to in vivo NMR on taurine-biological samples, we have developed the (33)S NMR cryogenic probe, which is applicable to taurine solutions. The NMR sensitivity gain relative to a conventional broadband probe is as large as 3.5. This work suggests that improvements in the preamplifier could allow NMR measurements on 100 microM taurine solutions, which is the level of sensitivity necessary for biological samples. PMID:19334961

  15. 13C CPMAS NMR study of the structure of p- t-butylcalix[4]arene-alkylbenzene clathrates in the solid state

    NASA Astrophysics Data System (ADS)

    Yamanobe, Takeshi; Nakamura, Ichinari; Hibino, Kazunobu; Komoto, Ta-i.; Kurosu, Hiromichi; Ando, Isao; Nakamoto, Yoshiaki; Ishida, Shin-ichiro

    1995-08-01

    The NMR chemical shifts measured by the 13C CPMAS method for clathrates of p- t-butylcalix[4]arenes including mobile alkylbenzenes in the solid state revealed that the alkyl groups of the guest molecules are located in the cavity of p- t-butylcalix[4]arene, depending on the size of the alkyl group. It was found that n-alkylbenzenes are easily incorporated into p- t-butylcalix[4]arene in the order of toluene > ethylbenzene ⩾ n-propylbenzene. For isopropylbenzene, the bulky isopropyl group prevents incorporation into the cavity of the host molecule.

  16. Crystal Structures and Vibrational and Solid-State (CPMAS) NMR Spectroscopy of Some Bis(triphenylphosphine)silver(I) Sulfate, Selenate and Phosphate Systems.

    SciTech Connect

    Bowmaker, Graham A.; Hanna, John V.; Rickard, Clifton E.; Lipton, Andrew S.

    2001-01-01

    The complexes [Ag2(PPh3)4EO4].2H2O(E=S, Se) (1,2), [Ag(PPh3)2HEO4].H2O (E=S, Se)(3,4) and [Ag9PPh3)2H2PO4].2EtOH (5) have been prepared and studied by X-ray crystallography and by infrared and solid-state 13C and 31 P cross-polarization, magic-angle-spinning (CPMAS) NMR spectroscopy.

  17. NMR probe for dynamic-angle spinning

    NASA Astrophysics Data System (ADS)

    Mueller, K. T.; Chingas, G. C.; Pines, A.

    1991-06-01

    We describe the design of a probe for dynamic-angle spinning (DAS) NMR experiments, comprised of a spinning cylindrical sample holder whose axis may be reoriented rapidly between discrete directions within the bore of a superconducting magnet. This allows the refocusing of nuclear spin magnetization that evolves under anisotropic interactions such as chemical shift anisotropy and quadrupolar coupling, providing high resolution NMR spectra for quadrupolar nuclei in solid materials. The probe includes an axial air delivery system to bearing and drive jets which support and spin a rotor containing the sample. Axis reorientation is accomplished with a pulley attached to the probehead and coupled to a stepping motor outside of the magnet. The choice of motor and gear ratio is based on an analysis of the moments of inertia of the motor and load, the desired angular resolution, and simplicity of design. Control of angular accuracy and precision are discussed, as well as the efficiency of radiofrequency irradiation and detection. High resolution DAS spectra of oxygen-17 and aluminum-27 nuclei in polycrystalline minerals illustrate the experimental capabilities.

  18. Insight into the conformational stability of membrane-embedded BamA using a combined solution and solid-state NMR approach.

    PubMed

    Sinnige, Tessa; Houben, Klaartje; Pritisanac, Iva; Renault, Marie; Boelens, Rolf; Baldus, Marc

    2015-04-01

    The β-barrel assembly machinery (BAM) is involved in folding and insertion of outer membrane proteins in Gram-negative bacteria, a process that is still poorly understood. With its 790 residues, BamA presents a challenge to current NMR methods. We utilized a "divide and conquer" approach in which we first obtained resonance assignments for BamA's periplasmic POTRA domains 4 and 5 by solution NMR. Comparison of these assignments to solid-state NMR (ssNMR) data obtained on two BamA constructs including the transmembrane domain and one or two soluble POTRA domains suggested that the fold of POTRA domain 5 critically depends on the interface with POTRA 4. Using specific labeling schemes we furthermore obtained ssNMR resonance assignments for residues in the extracellular loop 6 that is known to be crucial for BamA-mediated substrate folding and insertion. Taken together, our data provide novel insights into the conformational stability of membrane-embedded, non-crystalline BamA. PMID:25567766

  19. Three-dimensional structure determination of N-(p-Tolyl)-dodecylsulfonamide from powder diffraction data and validation of structure using solid-state NMR spectroscopy.

    PubMed

    Rajeswaran, Manju; Blanton, Thomas N; Zumbulyadis, Nicholas; Giesen, David J; Conesa-Moratilla, Carlota; Misture, Scott T; Stephens, Peter W; Huq, Ashfia

    2002-12-01

    The three-dimensional structure, conformation, and packing of molecules in the solid state are crucial components used in the optimization of many technologically useful materials properties. Single-crystal X-ray diffraction is the traditional and most effective method of determining 3-D structures in the solid state. Obtaining single crystals that are sufficiently large and free of imperfections is often laborious, time-consuming, and, occasionally, impossible. The feasibility of an integrated approach to the determination and verification of a complete three-dimensional structure for a medium-sized organic molecule without using single crystals is demonstrated for the case of an organic stabilizer compound N-(p-tolyl)-dodecylsulfonamide. The approach uses a combination of powder XRD data, several computational packages involving Monte Carlo simulations and ab initio quantum mechanical calculations, and experimental solid-state NMR chemical shifts. Structure elucidation of N-(p-tolyl)-dodecylsulfonamide revealed that the Bravais lattice is monoclinic, with cell dimensions of a = 38.773 A, b = 5.507 A, c = 9.509 A, and beta = 86.35 degrees, and a space group of P21/c. PMID:12452721

  20. Study of water dynamics in the soaking, steaming, and solid-state fermentation of glutinous rice by LF-NMR: a novel monitoring approach.

    PubMed

    Li, Teng; Tu, Chuanhai; Rui, Xin; Gao, Yangwen; Li, Wei; Wang, Kun; Xiao, Yu; Dong, Mingsheng

    2015-04-01

    Solid-state fermentation (SSF) of starchy grain is a traditional technique for food and alcoholic beverage production in East Asia. In the present study, low-field nuclear magnetic resonance (LF-NMR) was introduced for the elucidation of water dynamics and microstructure alternations during the soaking, steaming, and SSF of glutinous rice as a rapid real-time monitoring method. Three different proton fractions with different mobilities were identified based on the degree of interaction between biopolymers and water. Soaking and steaming significantly changed the proton distribution of the sample. The different phases of SSF were reflected by the T2 parameters. In addition, the variations in the T2 parameters were explained by the microstructure changes of rice induced by SSF. The fermentation time and T2 parameters were sigmoidally correlated. Thus, LF-NMR may be an effective real-time monitoring method for SSF in starch systems. PMID:25775016

  1. Solid-State NMR and DFT Studies on the Formation of Well-Defined Silica-Supported Tantallaaziridines: From Synthesis to Catalytic Application.

    PubMed

    Hamzaoui, Bilel; Pelletier, Jérémie D A; Abou-Hamad, Edy; Chen, Yin; El Eter, Mohamed; Chermak, Edrisse; Cavallo, Luigi; Basset, Jean-Marie

    2016-02-01

    Single-site, well-defined, silica-supported tantallaaziridine intermediates [≡Si-O-Ta(η(2) -NRCH2 )(NMe2 )2 ] [R=Me (2), Ph (3)] were prepared from silica-supported tetrakis(dimethylamido)tantalum [≡Si-O-Ta(NMe2 )4 ] (1) and fully characterized by FTIR spectroscopy, elemental analysis, and (1) H,(13) C HETCOR and DQ TQ solid-state (SS) NMR spectroscopy. The formation mechanism, by β-H abstraction, was investigated by SS NMR spectroscopy and supported by DFT calculations. The C-H activation of the dimethylamide ligand is favored for R=Ph. The results from catalytic testing in the hydroaminoalkylation of alkenes were consistent with the N-alkyl aryl amine substrates being more efficient than N-dialkyl amines. PMID:26875939

  2. 1H and 23Na MAS NMR spectroscopy of cationic species in CO2 selective alkaline earth metal porous silicoaluminophosphates prepared via liquid and solid state ion exchange

    NASA Astrophysics Data System (ADS)

    Arévalo-Hidalgo, Ana G.; Dugar, Sneha; Fu, Riqiang; Hernández-Maldonado, Arturo J.

    2012-07-01

    The location of extraframework cations in Sr2+ and Ba2+ ion-exchanged SAPO-34 was estimated by means of 1H and 23Na MAS NMR spectroscopy and spectral deconvolution. Incorporation of the alkaline earth metal cations onto the SAPO framework was achieved via liquid state ion exchange, coupled partial detemplation/solid-state ion exchange, and combination of both techniques. MAS NMR revealed that the level of ion exchange was limited by the presence of protons and sodium cations near hexagonal prisms (site SI), which are relatively difficult to exchange with the alkaline earth metal due to steric and charge repulsion criteria. In addition, the presence of ammonium cations in the supercages facilitated the exchange of otherwise tenacious hydrogen as corroborated by unit cell compositional data as well as enhanced CO2 adsorption at low partial pressures. The extraframework ammonium species were produced from partial detemplation of the structure-directing agent employed for the SAPO-34 synthesis, tetraethylammonium.

  3. Structural features of a bituminous coal and their changes during low-temperature oxidation and loss of volatiles investigated by advanced solid-state NMR spectroscopy

    USGS Publications Warehouse

    Mao, J.-D.; Schimmelmann, A.; Mastalerz, Maria; Hatcher, P.G.; Li, Y.

    2010-01-01

    Quantitative and advanced 13C solid-state NMR techniques were employed to investigate (i) the chemical structure of a high volatile bituminous coal, as well as (ii) chemical structural changes of this coal after evacuation of adsorbed gases, (iii) during oxidative air exposure at room temperature, and (iv) after oxidative heating in air at 75 ??C. The solid-state NMR techniques employed in this study included quantitative direct polarization/magic angle spinning (DP/MAS) at a high spinning speed of 14 kHz, cross polarization/total sideband suppression (CP/TOSS), dipolar dephasing, CH, CH2, and CHn selection, 13C chemical shift anisotropy (CSA) filtering, two-dimensional (2D) 1H-13C heteronuclear correlation NMR (HETCOR), and 2D HETCOR with 1H spin diffusion. With spectral editing techniques, we identified methyl CCH 3, rigid and mobile methylene CCH2C, methine CCH, quaternary Cq, aromatic CH, aromatic carbons bonded to alkyls, small-sized condensed aromatic moieties, and aromatic C-O groups. With direct polarization combined with spectral-editing techniques, we quantified 11 different types of functional groups. 1H-13C 2D HETCOR NMR experiments indicated spatial proximity of aromatic and alkyl moieties in cross-linked structures. The proton spin diffusion experiments indicated that the magnetization was not equilibrated at a 1H spin diffusion time of 5 ms. Therefore, the heterogeneity in spatial distribution of different functional groups should be above 2 nm. Recoupled C-H long-range dipolar dephasing showed that the fraction of large charcoal-like clusters of polycondensed aromatic rings was relatively small. The exposure of this coal to atmospheric oxygen at room temperature for 6 months did not result in obvious chemical structural changes of the coal, whereas heating at 75 ??C in air for 10 days led to oxidation of coal and generated some COO groups. Evacuation removed most volatiles and caused a significant reduction in aliphatic signals in its DP/MAS spectrum. DP/MAS, but not CP/MAS, allowed us to detect the changes during low-temperature oxidation and loss of volatiles. These results demonstrate the applicability of advanced solid-state NMR techniques in chemical characterization of coal. ?? 2010 American Chemical Society.

  4. Local environment and composition of magnesium gallium layered double hydroxides determined from solid-state {sup 1}H and {sup 71}Ga NMR spectroscopy

    SciTech Connect

    Petersen, Line Boisen; Lipton, Andrew S.; Zorin, Vadim; Nielsen, Ulla Gro

    2014-11-15

    Ordering of gallium(III) in a series of magnesium gallium (MgGa) layered double hydroxides (LDHs), [Mg{sub 1−x}Ga{sub x}(OH){sub 2}(NO{sub 3}){sub x}·yH{sub 2}O] was investigated using solid-state {sup 1}H and {sup 71}Ga NMR spectroscopy as well as powder X-ray diffraction. Three different proton environments from Mg{sub 3}-OH, Mg{sub 2}Ga-OH and intergallery water molecules were assigned and quantified using ({sup 1}H,{sup 71}Ga) HETCOR and {sup 1}H MAS NMR. A single {sup 71}Ga site originating from the unique Ga site in the MgGa LDH's was observed in {sup 71}Ga MAS and 3QMAS NMR spectra. Both {sup 1}H MAS NMR spectra recorded at 21.1 T (900 MHz) and elemental analysis show that the synthesized MgGa LDH's had a lower Mg:Ga ratio than that of the starting reactant solution. The origin of this is the formation of soluble [Ga(OH){sub 4}]{sup −} complexes formed during synthesis, and not due to formation of insoluble gallium (oxy)hydroxides. No sign of Ga-O-Ga connectivities or defects were detected for the MgGa LDH's. - Graphical abstract: Two types of hydroxides groups are observed in magnesium gallium layered double hydroxides revealing an ordering of Ga in the cation layer. - Highlights: • Ga is ordered in our magnesium gallium layered double hydroxides. • Ga depletion due to formation of soluble Ga complexes during synthesis. • No sign of Ga rich regions in magnesium gallium LDHs. • Solid state {sup 1}H and {sup 71}Ga give detailed insight into the structure.

  5. A General Assignment Method for Oriented Sample (OS) Solid-state NMR of Proteins Based on The Correlation of Resonances through Heteronuclear Dipolar Couplings in Samples Aligned Parallel and Perpendicular to the Magnetic Field

    PubMed Central

    Lu, George J.; Son, Woo Sung; Opella, Stanley J.

    2011-01-01

    A general method for assigning oriented sample (OS) solid-state NMR spectra of proteins is demonstrated. In principle, this method requires only a single sample of a uniformly 15N-labeled membrane protein in magnetically aligned bilayers, and a previously assigned isotropic chemical shift spectrum obtained either from solution NMR on micelle or isotropic bicelle samples or from magic angle spinning (MAS) solid-state NMR on unoriented proteoliposomes. The sequential isotropic resonance assignments are transferred to the OS solid-state NMR spectra of aligned samples by correlating signals from the same residue observed in protein-containing bilayers aligned with their normals parallel and perpendicular to the magnetic field. The underlying principle is that the resonances from the same residue have heteronuclear dipolar couplings that differ by exactly a factor of two between parallel and perpendicular alignments. The method is demonstrated on the membrane-bound form of Pf1 coat protein in phospholipid bilayers, whose assignments have been previously made using an earlier generation of methods that relied on the preparation of many selectively labeled (by residue type) samples. The new method provides the correct resonance assignments using only a single uniformly 15N-labeled sample, two solid-state NMR spectra, and a previously assigned isotropic spectrum. Significantly, this approach is equally applicable to residues in alpha helices, beta sheets, loops, and any other elements of tertiary structure. Moreover, the strategy bridges between OS solid-state NMR of aligned samples and solution NMR or MAS solid-state NMR of unoriented samples. In combination with the development of complementary experimental methods, it provides a step towards unifying these apparently different NMR approaches. PMID:21316275

  6. Structure determination of a bio-inspired self-assembled light-harvesting antenna by solid-state NMR and molecular modeling.

    PubMed

    Pandit, Anjali; Ocakoglu, Kasim; Buda, Francesco; van Marle, Thomas; Holzwarth, Alfred R; de Groot, Huub J M

    2013-09-26

    The molecular stacking of an artificial light-harvesting antenna self-assembled from 3(1)-aminofunctionalized zinc-chlorins was determined by solid-state NMR in combination with quantum-chemical and molecular-mechanics modeling. A library of trial molecular stacking arrangements was generated based on available structural data for natural and semisynthetic homologues of the Zn-chlorins. NMR assignments obtained for the monomer in solution were validated for self-assembled aggregates and refined with (1)H-(13)C heteronuclear correlation spectroscopy data collected from samples with (13)C at natural abundance. Solid-state ring-current shifts for the (1)H provided spatial constraints to determine the molecular overlap. This procedure allows for a discrimination between different self-assembled structures and a classification of the stacking mode in terms of electric dipole alignment and π-π interactions, parameters that determine the functional properties of light-harvesting assemblies and conducting nanowires. The combination with quantum-mechanical modeling then allowed building a low-resolution packing model in silico from molecular stacks. The method allows for moderate disorder and residual polymorphism at the stack or molecular level and is generally applicable to determine molecular packing structures of aromatic molecules with structural asymmetry, such as is commonly provided by functionalized side chains that serve to tune the self-assembly process. PMID:23566216

  7. Solid-state 13C NMR study of the mobility of polysaccharides in the cell walls of two apple cultivars of different firmness.

    PubMed

    Ng, Jovyn K T; Zujovic, Zoran D; Smith, Bronwen G; Johnston, Jason W; Schröder, Roswitha; Melton, Laurence D

    2014-03-11

    Solid-state (13)C nuclear magnetic resonance (NMR) was used to compare differences in mobility of the cell wall polysaccharides of 'Scifresh' and 'Royal Gala' apples after 20 weeks of storage. The texture of 'Scifresh' apples was markedly firmer than that of 'Royal Gala' at the end of storage. In a novel approach Two Pulse Phase Modulation (TPPM) decoupling was combined with cross polarisation (CP) and single pulse excitation (SPE) experiments. The resulting high resolution solid-state SPE spectra, unprecedented for apple cell walls, allowed a detailed insight into the physical and chemical properties of very mobile polysaccharides such as the arabinan and galactan side chains of the pectic polysaccharide rhamnogalacturonan I (RG-I). NMR showed that the cellulose rigidity was the same in the two cultivars, while arabinans were more mobile than galactans in both. Unexpectedly, arabinans in 'Scifresh' cell walls were more mobile than those in 'Royal Gala' which was unforeseen considering the greater firmness of the 'Scifresh' cultivar. PMID:24423413

  8. Nano ranged 1,3-dithia-2-stannacyclopentane(IV) alkyldithiocarbonates: solid-state NMR, powder X-ray diffraction and thermal decomposition studies

    NASA Astrophysics Data System (ADS)

    Chauhan, H. P. S.; Bakshi, Abhilasha; Joshi, Sapana; Carpenter, Jaswant

    2015-03-01

    The research work, performed during the course of present investigations is concerned with the synthesis, physico-chemical [elemental analysis and melting points determinations], spectral [IR, Far-IR, multinuclear (13C and 119Sn) solid-state NMR], powder XRD and thermal studies on 1,3-dithia-2-stannacyclopentane(IV) derivatives with alkyldithiocarbonates. The derivatives, [(CH2S)2Sn(S2COR)Cl] and [(CH2S)2Sn(S2COR)2] have been synthesized by the reaction of 1,3-dithia-2-stannacyclopentane(IV) dichloride with potassium alkyldithiocarbonates in 1:1 and 1:2 stoichiometry, respectively. Thermal decomposition study provides significant information about the formation of tin sulfide (SnS2) which has a number of commercial applications in various fields. Crystallite size of the derivatives has been found in nano range (∼132 nm) with the help of powder XRD studies. Sideband patterns obtained in our respective derivatives indicate distorted geometry and increased shielding of tin in solid state as compared to solution state 119Sn NMR, could be an indication of high electron density at tin atom.

  9. Elucidating the guest-host interactions and complex formation of praziquantel and cyclodextrin derivatives by (13)C and (15)N solid-state NMR spectroscopy.

    PubMed

    Arra, Eva C; Ferreira, M Joo G; Salomon, Claudio J; Nunes, Teresa G

    2015-12-30

    Praziquantel is the drug of choice to treat several parasitic infections including the neglected tropical disease schistosomiasis. Due to its low aqueous solubility, cyclodextrins have been tested as potential host candidates to prepare praziquantel inclusion complexes with improved solubility. For the first time, the interactions of praziquantel with ?-cyclodextrin and ?-cyclodextrin derivatives (methyl-?-cyclodextrin and hydroxypropyl-?-cyclodextrin) were investigated using high resolution solid-state NMR spectroscopy. The results of this work confirmed that solid-state NMR experiments provided structural characterization, demonstrating the formation of inclusion complexes most probably with PZQ adopting an anti conformation, also the most likely in amorphous raw PZQ. Further information on the interaction of praziquantel with methyl-?-cyclodextrin was obtained from proton rotating-frame relaxation time measurements, sensitive to kilohertz-regime motions but modulated by spin-diffusion. Evidences were presented in all cases for praziquantel complexation through the aromatic ring. In addition, 1:2 drug:carrier molar ratio appears to be the most probable and therefore suitable stoichiometry to improve pharmaceutical formulations of this antischistosomal drug. PMID:26602291

  10. Local environment and composition of magnesium gallium layered double hydroxides determined from solid-state 1H and 71Ga NMR spectroscopy

    NASA Astrophysics Data System (ADS)

    Petersen, Line Boisen; Lipton, Andrew S.; Zorin, Vadim; Nielsen, Ulla Gro

    2014-11-01

    Ordering of gallium(III) in a series of magnesium gallium (MgGa) layered double hydroxides (LDHs), [Mg1-xGax(OH)2(NO3)x·yH2O] was investigated using solid-state 1H and 71Ga NMR spectroscopy as well as powder X-ray diffraction. Three different proton environments from Mg3sbnd OH, Mg2Gasbnd OH and intergallery water molecules were assigned and quantified using {1H,71Ga} HETCOR and 1H MAS NMR. A single 71Ga site originating from the unique Ga site in the MgGa LDH's was observed in 71Ga MAS and 3QMAS NMR spectra. Both 1H MAS NMR spectra recorded at 21.1 T (900 MHz) and elemental analysis show that the synthesized MgGa LDH's had a lower Mg:Ga ratio than that of the starting reactant solution. The origin of this is the formation of soluble [Ga(OH)4]- complexes formed during synthesis, and not due to formation of insoluble gallium (oxy)hydroxides. No sign of Gasbnd Osbnd Ga connectivities or defects were detected for the MgGa LDH's.

  11. Helical jump motions of poly(L-lactic acid) chains in the α phase as revealed by solid-state NMR.

    PubMed

    Chen, Wei; Reichert, Detlef; Miyoshi, Toshikazu

    2015-03-26

    The molecular dynamics of Poly(L-lactic Acid) (PLLA) chains in the α phase was investigated by Solid-State NMR spectroscopy. (13)C high-resolution NMR clearly indicates that the crystalline signals split into 2, 3, and 4 signals for the CH3, CH and CO groups, respectively at 25 °C, while the amorphous signals give a broad component at the bottom of the crystalline signals. (13)C NMR spectra show that the crystalline line shape changes with increasing temperatures well above the glass transition temperature (Tg) and imply the presence of the molecular dynamics in the crystalline region. Comparisons of the evolution-time dependence of CODEX data and simulation results based on reorientation of chemical shift anisotropy (CSA) indicate that the chains in the α phase perform helical jump motions in the slow dynamic range at temperatures above 115 °C. The mixing-time dependence of the CODEX data yields an activation energy of Ea of (95 ± 8) kJ/mol for the helical jump motions. Moreover, two-dimensional exchange NMR with highly resolved signals for the CO group provides cross peaks among four well resolved signals due to the helical jumps. Comparison of 2D buildup curves of the cross peaks and calculated data determines that helical jump motions prefer largely uncorrelated random back-and-forth motions between the neighboring sites, possibly enabling large-scale chain diffusion in the crystalline regions. PMID:25730501

  12. Investigation of the Conversion Reaction Mechanisms for Binary Copper(II) Compounds by Solid-State NMR Spectroscopy and X-ray Diffraction

    SciTech Connect

    Yamakawa, N.; Jiang, M; Grey, C

    2009-01-01

    The conversion reaction mechanisms of CuS, CuF2, and CuO during the electrochemical reaction with Li are studied by solid-state 63Cu, 19F, and 7Li nuclear magnetic resonance (NMR) and X-ray diffraction (XRD). For CuS, a two-step reaction is observed that is associated with an insertion reaction involving first limited incorporation of Li into CuS and then a two-phase reaction to form a material with the approximate composition LiCuS. This is followed by a conversion reaction to form Li2S and Cu, Cu1.96S being formed as a side product of the decomposition of LiCuS. Evidence for the insertion phases is found from both NMR and XRD. A direct conversion reaction to form LiF and Cu is seen for CuF2, whereas the 7Li NMR results indicate that CuO can tolerate a small amount of Li substitution before reacting to form Li2O and Cu. Both the diffraction and NMR results indicate that the size of the Cu particles formed on discharge are much larger in the CuS system, which is thought to result from the higher Cu1+ mobilities in the intermediate intercalation compounds LixCuS. The factors that control the possible mechanisms for these conversion reactions are discussed.

  13. High-resolution structure of a Shigella type III secretion needle by solid-state NMR and cryo-electron microscopy

    PubMed Central

    Demers, Jean-Philippe; Habenstein, Birgit; Loquet, Antoine; Vasa, Suresh Kumar; Giller, Karin; Becker, Stefan; Baker, David; Lange, Adam; Sgourakis, Nikolaos G.

    2014-01-01

    We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state NMR (ssNMR) chemical shifts and distance constraints define the local secondary structure, protein fold and inter-subunit interactions. Finally, Rosetta structure calculations provide a general framework to integrate the different sources of structural information. Combining a 7.7-Å cryo-EM density map and 996 ssNMR distance constraints, the structure of the Type-III Secretion System (T3SS) needle of Shigella flexneri is determined to a precision of 0.4 Å. The calculated structures are cross-validated using an independent dataset of 691 ssNMR constraints and STEM measurements. The hybrid model resolves the conformation of the non-conserved N-terminus, that occupies a protrusion in the cryo-EM density, and reveals conserved pore residues forming a continuous pattern of electrostatic interactions, thereby suggesting a mechanism for effector protein translocation. PMID:25264107

  14. (13)C solid-state NMR analysis of the most common pharmaceutical excipients used in solid drug formulations Part II: CP kinetics and relaxation analysis.

    PubMed

    Pisklak, Dariusz Maciej; Zielińska-Pisklak, Monika; Szeleszczuk, Łukasz; Wawer, Iwona

    2016-04-15

    Excipients used in the solid drug formulations differ in their NMR relaxation and (13)C cross-polarization (CP) kinetics parameters. Therefore, experimental parameters like contact time of cross-polarization and repetition time have a major impact on the registered solid state NMR spectra and in consequence on the results of the NMR analysis. In this work the CP kinetics and relaxation of the most common pharmaceutical excipients: anhydrous α-lactose, α-lactose monohydrate, mannitol, sucrose, sorbitol, sodium starch glycolate type A and B, starch of different origin, microcrystalline cellulose, hypromellose, ethylcellulose, methylcellulose, hydroxyethylcellulose, sodium alginate, magnesium stearate, sodium laurilsulfate and Kollidon(®) were analyzed. The studied excipients differ significantly in their optimum repetition time (from 5s to 1200s) and T1ρ(I) parameters (from 2ms to 73ms). The practical use of those differences in the excipients composition analysis was demonstrated on the example of commercially available tablets containing indapamide as an API. The information presented in this article will help to choose the correct acquisition parameters and also will save the time and effort needed for their optimization in the NMR analysis of the solid drug formulations. PMID:26836362

  15. Solid-state 27Al and 29Si NMR characterization of hydrates formed in calcium aluminate silica fume mixtures

    NASA Astrophysics Data System (ADS)

    Pena, P.; Rivas Mercury, J. M.; de Aza, A. H.; Turrillas, X.; Sobrados, I.; Sanz, J.

    2008-08-01

    Partially deuterated Ca 3Al 2(SiO 4) y(OH) 12-4y-Al(OH) 3 mixtures, prepared by hydration of Ca 3Al 2O 6 (C 3A), Ca 12Al 14O 33 (C 12A 7) and CaAl 2O 4 (CA) phases in the presence of silica fume, have been characterized by 29Si and 27Al magic-angle spinning-nuclear magnetic resonance (MAS-NMR) spectroscopies. NMR spectroscopy was used to characterize anhydrous and fully hydrated samples. In hydrated compounds, Ca 3Al 2(OH) 12 and Al(OH) 3 phases were detected. From the quantitative analysis of 27Al NMR signals, the Al(OH) 3/Ca 3Al 2(OH) 12 ratio was deduced. The incorporation of Si into the katoite structure, Ca 3Al 2(SiO 4) 3-x(OH) 4x, was followed by 27Al and 29Si NMR spectroscopies. Si/OH ratios were determined from the quantitative analysis of 27Al MAS-NMR components associated with Al(OH) 6 and Al(OSi)(OH) 5 environments. The 29Si NMR spectroscopy was also used to quantify the unreacted silica and amorphous calcium aluminosilicate hydrates formed, C-S-H and C-A-S-H for short. From 29Si NMR spectra, the amount of Si incorporated into different phases was estimated. Si and Al concentrations, deduced by NMR, transmission electron microscopy, energy dispersive spectrometry, and Rietveld analysis of both X-ray and neutron data, indicate that only a part of available Si is incorporated in katoite structures.

  16. Effect of iron content on the structure and disorder of iron-bearing sodium silicate glasses: A high-resolution 29Si and 17O solid-state NMR study

    NASA Astrophysics Data System (ADS)

    Kim, Hyo-Im; Sur, Jung Chul; Lee, Sung Keun

    2016-01-01

    Despite its geochemical importance and implications for the properties of natural magmatic melts, understanding the detailed structure of iron-bearing silicate glasses remains among the outstanding problems in geochemistry. This is mainly because solid-state NMR techniques, one of the most versatile experimental methods to probe the structure of oxide glasses, cannot be fully utilized for exploring the structural details of iron-bearing glasses as the unpaired electrons in Fe induce strong local magnetic fields that mask the original spectroscopic features (i.e., paramagnetic effect). Here, we report high-resolution 29Si and 17O solid-state NMR spectra of iron-bearing sodium silicate glasses (Na2O-Fe2O3-SiO2, Fe3+/ΣFe = 0.89 ± 0.04, thus containing both ferric and ferrous iron) with varying XFe2O3 [=Fe2O3/(Na2O + Fe2O3)], containing up to 22.9 wt% Fe2O3. This compositional series involves Fe-Na substitution at constant SiO2 contents of 66.7 mol% in the glasses. For both nuclides, the NMR spectra exhibit a decrease in the signal intensities and an increase in the peak widths with increasing iron concentration partly because of the paramagnetic effect. Despite the intrinsic difficulties that result from the pronounced paramagnetic effect, the 29Si and 17O NMR results yield structural details regarding the effect of iron content on Q speciation, spatial distribution of iron, and the extent of polymerization in the iron-bearing silicate glasses. The 29Si NMR spectra show an apparent increase in highly polymerized Q species with increasing XFe2O3 , suggesting an increase in the degree of melt polymerization. The 17O 3QMAS NMR spectra exhibit well-resolved non-bridging oxygen (NBO, Na-O-Si) and bridging oxygen (BO, Si-O-Si) peaks with varying iron concentration. By replacing Na2O with Fe2O3 (and thus with increasing iron content), the fraction of Na-O-Si decreases. Quantitative consideration of this effect confirms that the degree of polymerization is likely to increase with iron content and that Fe3+ is predominantly a network-former. The 17O NMR spectra suggest a moderate degree of preferential partitioning of iron between NBO and BO clusters. The present results bear strong promise for studying iron-bearing silicate glasses using solid-state NMR techniques, constraining the effect of iron content on the degree of polymerization. The observed changes in atomic structures of iron-bearing sodium silicate glasses will be helpful for unraveling atomic origins of the properties of natural silicate melts.

  17. YPdSn and YPd{sub 2}Sn: Structure, {sup 89}Y solid state NMR and {sup 119}Sn Moessbauer spectroscopy

    SciTech Connect

    Hoeting, Christoph; Eckert, Hellmut; Langer, Thorsten; Schellenberg, Inga; Poettgen, Rainer

    2012-06-15

    The stannides YPdSn and YPd{sub 2}Sn were synthesized by high-frequency melting of the elements in sealed tantalum tubes. Both structures were refined on the basis of single crystal X-ray diffractometer data: TiNiSi type, Pnma, a=715.4(1), b=458.8(1), c=789.1(1) pm, wR2=0.0461, 510 F{sup 2} values, 20 variables for YPdSn and MnCu{sub 2}Al type, Fm3 Macron m, a=671.44(8), wR2=0.0740, 55 F{sup 2} values, 5 parameters for YPd{sub 2}Sn. The yttrium atoms in the new stannide YPdSn are coordinated by two tilted Pd{sub 3}Sn{sub 3} hexagons (ordered AlB{sub 2} superstructure). In the Heusler phase YPd{sub 2}Sn each yttrium atom has octahedral tin coordination and additionally eight palladium neighbors. The cubic site symmetry of yttrium is reflected in the {sup 119}Sn Moessbauer spectrum which shows no quadrupole splitting. In contrast, YPdSn shows a single signal at {delta}=1.82(1) mm/s subjected to quadrupole splitting of {Delta}E{sub Q}=0.93(1) mm/s. Both compounds have been characterized by high-resolution {sup 89}Y solid state NMR spectroscopy, which indicates the presence of strong Knight shifts. The spectrum of YPd{sub 2}Sn is characterized by an unusually large linewidth, suggesting the presence of a Knight shift distribution reflecting local disordering effects. The range of {sup 89}Y Knight shifts of several binary and ternary intermetallic yttrium compounds is briefly discussed. - Graphical abstract: YPdSn and YPd{sub 2}Sn: Structure, {sup 89}Y solid state NMR and {sup 119}Sn Moessbauer spectroscopy. Highlights: Black-Right-Pointing-Pointer Synthesis and structure of ternary stannides YPdSn and YPd{sub 2}Sn. Black-Right-Pointing-Pointer {sup 119}Sn Moessbauer spectroscopic investigation of YPdSn and YPd{sub 2}Sn. Black-Right-Pointing-Pointer {sup 89}Y solid state NMR of intermetallics.

  18. {sup 29}Si solid state NMR investigation of pozzolanic reaction occurring in lime-treated Ca-bentonite

    SciTech Connect

    Pomakhina, Elena; Deneele, Dimitri; Gaillot, Anne-Claire; Paris, Michael; Ouvrard, Guy

    2012-04-15

    Lime is widely used as additive to improve the mechanical properties of natural soil used in earthworks. However, the physico-chemical mechanisms involved are yet not well understood. In order to develop and optimize this treatment method, a better understanding of the interaction between lime and the minerals of the soils, in particular clay minerals, is required. In this study, Ca-bentonite was treated with 2, 5 and 10 wt.% of lime during 1 to 98 days. Modifications in the Si local environment were then monitored by solid state nuclear magnetic resonance to investigate the pozzolanic reaction. All the soil mineral phases contribute to the release of Si and to the pozzolanic reaction, with a rapid and total consumption of Si-polymorph and an exacerbated dissolution of montmorillonite. Mechanism of C-S-H formation, function of the Ca content in the system, was found to match the sorosilicate-tobermorite model described in cement systems.

  19. Solid-state 13C NMR studies of a large fossil gymnosperm from the Yallourn Open Cut, Latrobe Valley, Australia

    USGS Publications Warehouse

    Bates, A.L.; Hatcher, P.G.

    1989-01-01

    A series of samples taken from the cross section of a 3-m-diameter fossilized gymnospermous log (Araucariaceae) in the Yallourn Seam of the Australian brown coals was examined by solid state 13C nuclear magnetic resonance to delineate chemical changes related to the combined processes of peatification and coalification. The results show that cellulosic materials were degraded and lost on the periphery of the log, however, the degree of such degradation in the central core is substantially less. The lignin is uniformly altered by coalification reactions to a macromolecular substance displaying decreased aryl ether linkages but significantly greater amounts of carbon linkages compared to modern lignin. Changes in the methoxyl carbon contents of lignin in cross section reveal demethylation reactions, but these do not appear to be related to degree of carbon linking. Both the degredation of cellulosic materials and demethylation of lignin appear to be early diagenetic processes occurring during peatification independently of the coalification reactions. ?? 1989.

  20. Solid-state (185/187)Re NMR and GIPAW DFT study of perrhenates and Re2(CO)10: chemical shift anisotropy, NMR crystallography, and a metal-metal bond.

    PubMed

    Widdifield, Cory M; Perras, Frédéric A; Bryce, David L

    2015-04-21

    Advances in solid-state nuclear magnetic resonance (SSNMR) methods, such as dynamic nuclear polarization (DNP), intricate pulse sequences, and increased applied magnetic fields, allow for the study of systems which even very recently would be impractical. However, SSNMR methods using certain quadrupolar probe nuclei (i.e., I > 1/2), such as (185/187)Re remain far from fully developed due to the exceedingly strong interaction between the quadrupole moment of these nuclei and local electric field gradients (EFGs). We present a detailed high-field (B0 = 21.1 T) experimental SSNMR study on several perrhenates (KReO4, AgReO4, Ca(ReO4)2·2H2O), as well as ReO3 and Re2(CO)10. We propose solid ReO3 as a new rhenium SSNMR chemical shift standard due to its reproducible and sharp (185/187)Re NMR resonances. We show that for KReO4, previously poorly understood high-order quadrupole-induced effects (HOQIE) on the satellite transitions can be used to measure the EFG tensor asymmetry (i.e., ηQ) to nearly an order-of-magnitude greater precision than competing SSNMR and nuclear quadrupole resonance (NQR) approaches. Samples of AgReO4 and Ca(ReO4)2·2H2O enable us to comment on the effects of counter-ions and hydration upon Re(vii) chemical shifts. Calcium-43 and (185/187)Re NMR tensor parameters allow us to conclude that two proposed crystal structures for Ca(ReO4)2·2H2O, which would be considered as distinct, are in fact the same structure. Study of Re2(CO)10 provides insights into the effects of Re-Re bonding on the rhenium NMR tensor parameters and rhenium oxidation state on the Re chemical shift value. As overtone NQR experiments allowed us to precisely measure the (185/187)Re EFG tensor of Re2(CO)10, we were able to measure rhenium chemical shift anisotropy (CSA) for the first time in a powdered sample. Experimental observations are supported by gauge-including projector augmented-wave (GIPAW) density functional theory (DFT) calculations, with NMR tensor calculations also provided for NH4ReO4, NaReO4 and RbReO4. These calculations are able to reproduce many of the experimental trends in rhenium δiso values and EFG tensor magnitudes. Using KReO4 as a prototypical perrhenate-containing system, we establish a correlation between the tetrahedral shear strain parameter (|ψ|) and the nuclear electric quadrupolar coupling constant (CQ), which enables the refinement of the structure of ND4ReO4. Shortcomings in traditional DFT approaches, even when including relativistic effects via the zeroth-order regular approximation (ZORA), for calculating rhenium NMR tensor parameters are identified for Re2(CO)10. PMID:25790263

  1. Solid state 13C NMR analysis of shales and coals from Laramide Basins. Final report, March 1, 1995--March 31, 1996

    SciTech Connect

    Miknis, F.P.; Jiao, Z.S.; Zhao, Hanqing; Surdam, R.C.

    1998-12-31

    This Western Research Institute (WRI) jointly sponsored research (JSR) project augmented and complemented research conducted by the University of Wyoming Institute For Energy Research for the Gas Research Institute. The project, {open_quotes}A New Innovative Exploitation Strategy for Gas Accumulations Within Pressure Compartments,{close_quotes} was a continuation of a project funded by the GRI Pressure Compartmentalization Program that began in 1990. That project, {open_quotes}Analysis of Pressure Chambers and Seals in the Powder River Basin, Wyoming and Montana,{close_quotes} characterized a new class of hydrocarbon traps, the discovery of which can provide an impetus to revitalize the domestic petroleum industry. In support of the UW Institute For Energy Research`s program on pressure compartmentalization, solid-state {sup 13}C NMR measurements were made on sets of shales and coals from different Laramide basins in North America. NMR measurements were made on samples taken from different formations and depths of burial in the Alberta, Bighorn, Denver, San Juan, Washakie, and Wind River basins. The carbon aromaticity determined by NMR was shown to increase with depth of burial and increased maturation. In general, the NMR data were in agreement with other maturational indicators, such as vitrinite reflectance, illite/smectite ratio, and production indices. NMR measurements were also obtained on residues from hydrous pyrolysis experiments on Almond and Lance Formation coals from the Washakie Basin. These data were used in conjunction with mass and elemental balance data to obtain information about the extent of carbon aromatization that occurs during artificial maturation. The data indicated that 41 and 50% of the original aliphatic carbon in the Almond and Lance coals, respectively, aromatized during hydrous pyrolysis.

  2. High-resolution structure of the Shigella type-III secretion needle by solid-state NMR and cryo-electron microscopy.

    PubMed

    Demers, Jean-Philippe; Habenstein, Birgit; Loquet, Antoine; Kumar Vasa, Suresh; Giller, Karin; Becker, Stefan; Baker, David; Lange, Adam; Sgourakis, Nikolaos G

    2014-01-01

    We introduce a general hybrid approach for determining the structures of supramolecular assemblies. Cryo-electron microscopy (cryo-EM) data define the overall envelope of the assembly and rigid-body orientation of the subunits while solid-state nuclear magnetic resonance (ssNMR) chemical shifts and distance constraints define the local secondar