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Sample records for allosteric signal transmission

  1. Molecular Dynamics Investigation of a Mechanism of Allosteric Signal Transmission in Ribosomes.

    PubMed

    Makarov, G I; Golovin, A V; Sumbatyan, N V; Bogdanov, A A

    2015-08-01

    The ribosome is a molecular machine that synthesizes all cellular proteins via translation of genetic information encoded in polynucleotide chain of messenger RNA. Transition between different stages of the ribosome working cycle is strictly coordinated by changes in structure and mutual position both of subunits of the ribosome and its ligands. Therein, information regarding structural transformations is transmitted between functional centers of the ribosome through specific signals. Usually, functional centers of ribosomes are located at a distance reaching up to several tens of angstroms, and it is believed that such signals are transduced allosterically. In our study, we attempted to answer the question of how allosteric signal can be transmitted from one of the so-called sensory elements of ribosomal tunnel (RT) to the peptidyl transferase center (PTC). A segment of RT wall from the E. coli ribosome composed of nucleotide residues A2058, A2059, m(2)A2503, G2061, A2062, and C2063 of its 23S rRNA was examined by molecular dynamics simulations. It was found that a potential signal transduction pathway A2058-C2063 acted as a dynamic ensemble of interdependent conformational states, wherein cascade-like changes can occur. It was assumed that structural rearrangement in the A2058-C2063 RT segment results in reversible inactivation of PTC due to a strong stacking contact between functionally important U2585 residue of the PTC and nucleotide residue C2063. A potential role for the observed conformational transition in the A2058-C2063 segment for regulating ribosome activity is discussed.

  2. The LD loop as an important structural element required for transmission of the allosteric signal in the HtrA (DegP) protease from Escherichia coli.

    PubMed

    Figaj, Donata; Gieldon, Artur; Bartczak, Marlena; Koper, Tomasz; Zarzecka, Urszula; Lesner, Adam; Lipinska, Barbara; Skorko-Glonek, Joanna

    2016-09-01

    High-temperature requirement A (HtrA; DegP) from Escherichia coli, an important element of the extracytoplasmic protein quality-control system, is a member of the evolutionarily conserved family of serine proteases. The characteristic feature of this protein is its allosteric mode of activation. The regulatory loops, L3, L2, L1 and LD, play a crucial role in the transmission of the allosteric signal. Yet, the role of LD has not been fully elucidated. Therefore, we undertook a study to explain the role of the individual LD residues in inducing and maintaining the proteolytic activity of HtrA. We investigated the influence of amino acid substitutions located within the LD loop on the kinetics of a model substrate cleavage as well as on the dynamics of the oligomeric structure of HtrA. We found that the mutations that were expected to disturb the loop's structure and/or interactions with the remaining regulatory loops severely diminished the proteolytic activity of HtrA. The opposite effect, that is, increased activity, was observed for G174S substitution, which was predicted to strengthen the interactions mediated by LD. HtrAG174S protein had an equilibrium shifted toward the active enzyme and formed preferentially high-order oligomeric forms.

  3. Signal peptides are allosteric activators of the protein translocase

    PubMed Central

    Gouridis, Giorgos; Karamanou, Spyridoula; Gelis, Ioannis; Kalodimos, Charalampos G.; Economou, Anastassios

    2010-01-01

    Extra-cytoplasmic polypeptides are usually synthesized as “preproteins” carrying aminoterminal, cleavable signal peptides1 and secreted across membranes by translocases. The main bacterial translocase comprises the SecYEG protein-conducting channel and the peripheral ATPase motor SecA2,3. Most proteins destined for the periplasm and beyond are exported post-translationally by SecA2,3. Preprotein targeting to SecA is thought to involve signal peptides4 and chaperones like SecB5,6. Here we reveal that signal peptides have a novel role beyond targeting: they are essential allosteric activators of the translocase. Upon docking on their binding groove on SecA, signal peptides act in trans to drive three successive states: first, “triggering” that drives the translocase to a lower activation energy state; then “trapping” that engages non-native preprotein mature domains docked with high affinity on the secretion apparatus and, finally, “secretion” during which trapped mature domains undergo multiple turnovers of translocation in segments7. A significant contribution by mature domains renders signal peptides less critical in bacterial secretory protein targeting than currently assumed. Rather, it is their function as allosteric activators of the translocase that renders signal peptides essential for protein secretion. A role for signal peptides and targeting sequences as allosteric activators may be universal in protein translocases. PMID:19924216

  4. AIM for Allostery: Using the Ising Model to Understand Information Processing and Transmission in Allosteric Biomolecular Systems.

    PubMed

    LeVine, Michael V; Weinstein, Harel

    2015-05-01

    In performing their biological functions, molecular machines must process and transmit information with high fidelity. Information transmission requires dynamic coupling between the conformations of discrete structural components within the protein positioned far from one another on the molecular scale. This type of biomolecular "action at a distance" is termed allostery. Although allostery is ubiquitous in biological regulation and signal transduction, its treatment in theoretical models has mostly eschewed quantitative descriptions involving the system's underlying structural components and their interactions. Here, we show how Ising models can be used to formulate an approach to allostery in a structural context of interactions between the constitutive components by building simple allosteric constructs we termed Allosteric Ising Models (AIMs). We introduce the use of AIMs in analytical and numerical calculations that relate thermodynamic descriptions of allostery to the structural context, and then show that many fundamental properties of allostery, such as the multiplicative property of parallel allosteric channels, are revealed from the analysis of such models. The power of exploring mechanistic structural models of allosteric function in more complex systems by using AIMs is demonstrated by building a model of allosteric signaling for an experimentally well-characterized asymmetric homodimer of the dopamine D2 receptor.

  5. CB(1) receptor allosteric modulators display both agonist and signaling pathway specificity.

    PubMed

    Baillie, Gemma L; Horswill, James G; Anavi-Goffer, Sharon; Reggio, Patricia H; Bolognini, Daniele; Abood, Mary E; McAllister, Sean; Strange, Phillip G; Stephens, Gary J; Pertwee, Roger G; Ross, Ruth A

    2013-02-01

    We have previously identified allosteric modulators of the cannabinoid CB(1) receptor (Org 27569, PSNCBAM-1) that display a contradictory pharmacological profile: increasing the specific binding of the CB(1) receptor agonist [(3)H]CP55940 but producing a decrease in CB(1) receptor agonist efficacy. Here we investigated the effect one or both compounds in a broad range of signaling endpoints linked to CB(1) receptor activation. We assessed the effect of these compounds on CB(1) receptor agonist-induced [(35)S]GTPγS binding, inhibition, and stimulation of forskolin-stimulated cAMP production, phosphorylation of extracellular signal-regulated kinases (ERK), and β-arrestin recruitment. We also investigated the effect of these allosteric modulators on CB(1) agonist binding kinetics. Both compounds display ligand dependence, being significantly more potent as modulators of CP55940 signaling as compared with WIN55212 and having little effect on [(3)H]WIN55212 binding. Org 27569 displays biased antagonism whereby it inhibits: agonist-induced guanosine 5'-O-(3-[(35)S]thio)triphosphate ([(35)S]GTPγS) binding, simulation (Gα(s)-mediated), and inhibition (Gα(i)-mediated) of cAMP production and β-arrestin recruitment. In contrast, it acts as an enhancer of agonist-induced ERK phosphorylation. Alone, the compound can act also as an allosteric agonist, increasing cAMP production and ERK phosphorylation. We find that in both saturation and kinetic-binding experiments, the Org 27569 and PSNCBAM-1 appeared to influence only orthosteric ligand maximum occupancy rather than affinity. The data indicate that the allosteric modulators share a common mechanism whereby they increase available high-affinity CB(1) agonist binding sites. The receptor conformation stabilized by the allosterics appears to induce signaling and also selectively traffics orthosteric agonist signaling via the ERK phosphorylation pathway.

  6. Convergent Transmission of RNAi Guide-Target Mismatch Information across Argonaute Internal Allosteric Network

    PubMed Central

    Joseph, Thomas T.; Osman, Roman

    2012-01-01

    In RNA interference, a guide strand derived from a short dsRNA such as a microRNA (miRNA) is loaded into Argonaute, the central protein in the RNA Induced Silencing Complex (RISC) that silences messenger RNAs on a sequence-specific basis. The positions of any mismatched base pairs in an miRNA determine which Argonaute subtype is used. Subsequently, the Argonaute-guide complex binds and silences complementary target mRNAs; certain Argonautes cleave the target. Mismatches between guide strand and the target mRNA decrease cleavage efficiency. Thus, loading and silencing both require that signals about the presence of a mismatched base pair are communicated from the mismatch site to effector sites. These effector sites include the active site, to prevent target cleavage; the binding groove, to modify nucleic acid binding affinity; and surface allosteric sites, to control recruitment of additional proteins to form the RISC. To examine how such signals may be propagated, we analyzed the network of internal allosteric pathways in Argonaute exhibited through correlations of residue-residue interactions. The emerging network can be described as a set of pathways emanating from the core of the protein near the active site, distributed into the bulk of the protein, and converging upon a distributed cluster of surface residues. Nucleotides in the guide strand “seed region” have a stronger relationship with the protein than other nucleotides, concordant with their importance in sequence selectivity. Finally, any of several seed region guide-target mismatches cause certain Argonaute residues to have modified correlations with the rest of the protein. This arises from the aggregation of relatively small interaction correlation changes distributed across a large subset of residues. These residues are in effector sites: the active site, binding groove, and surface, implying that direct functional consequences of guide-target mismatches are mediated through the cumulative

  7. Global Low Frequency Protein Motions in Long-Range Allosteric Signaling

    NASA Astrophysics Data System (ADS)

    McLeish, Tom; Rogers, Thomas; Townsend, Philip; Burnell, David; Pohl, Ehmke; Wilson, Mark; Cann, Martin; Richards, Shane; Jones, Matthew

    2015-03-01

    We present a foundational theory for how allostery can occur as a function of low frequency dynamics without a change in protein structure. Elastic inhomogeneities allow entropic ``signalling at a distance.'' Remarkably, many globular proteins display just this class of elastic structure, in particular those that support allosteric binding of substrates (long-range co-operative effects between the binding sites of small molecules). Through multi-scale modelling of global normal modes we demonstrate negative co-operativity between the two cAMP ligands without change to the mean structure. Crucially, the value of the co-operativity is itself controlled by the interactions around a set of third allosteric ``control sites.'' The theory makes key experimental predictions, validated by analysis of variant proteins by a combination of structural biology and isothermal calorimetry. A quantitative description of allostery as a free energy landscape revealed a protein ``design space'' that identified the key inter- and intramolecular regulatory parameters that frame CRP/FNR family allostery. Furthermore, by analyzing naturally occurring CAP variants from diverse species, we demonstrate an evolutionary selection pressure to conserve residues crucial for allosteric control. The methodology establishes the means to engineer allosteric mechanisms that are driven by low frequency dynamics.

  8. Allosteric mechanisms of G protein coupled receptor signaling: a structural perspective

    PubMed Central

    Thaker, Tarjani M.; Kaya, Ali I.; Preininger, Anita M.; Hamm, Heidi E.; Iverson, T.M.

    2012-01-01

    G protein-Coupled Receptors (GPCRs) use a complex series of intramolecular conformational changes to couple agonist binding to the binding and activation of cognate heterotrimeric G protein (Gαβγ). The mechanisms underlying this long-range activation have been identified using a variety of biochemical and structural approaches and have primarily used visual signal transduction via the GPCR rhodopsin and cognate heterotrimeric G protein transducin (Gt) as a model system. In this chapter, we will review the methods that have revealed allosteric signaling through rhodopsin and transducin. These methods can be applied to a variety of GPCR-mediated signaling pathways. PMID:22052489

  9. Differential pathway coupling efficiency of the activated insulin receptor drives signaling selectivity by xmeta, an allosteric partial agonist antibody

    Technology Transfer Automated Retrieval System (TEKTRAN)

    XMetA, an anti-insulin receptor (IR) monoclonal antibody, is an allosteric partial agonist of the IR. We have previously reported that XMetA activates the “metabolic-biased” Akt kinase signaling pathway while having little or no effect on the “mitogenic” MAPK signaling pathwayof ERK 1/2. To inves...

  10. Time scale independent signal transmission

    NASA Astrophysics Data System (ADS)

    Faltin, L.

    1980-05-01

    The paper presents a method which permits the conversion of time scale variations occurring during signal transmission into time shifts proportionally related to these variations. It is demonstrated that the method can be used to reject the adverse effects of the time scale variations (such as wow and flutter in magnetic tape recordings) and/or to determine the scale change exactly (such as would be required in Doppler signal processing). Finally, it is noted that since the system performance degrades with rising frequency of the time scale distortions, an upper bound for this frequency is derived.

  11. Assembly of the Sos1-Grb2-Gab1 Ternary Signaling Complex Is Under Allosteric Control

    PubMed Central

    McDonald, Caleb B.; Seldeen, Kenneth L.; Deegan, Brian J.; Bhat, Vikas; Farooq, Amjad

    2009-01-01

    Allostery has evolved as a form of local communication between interacting protein partners allowing them to quickly sense changes in their immediate vicinity in response to external cues. Herein, using isothermal titration calorimetry (ITC) in conjunction with circular dichroism (CD) and macromolecular modeling (MM), we show that the binding of Grb2 adaptor — a key signaling molecule involved in the activation of Ras GTPase — to its downstream partners Sos1 guanine nucleotide exchange factor and Gab1 docker is under tight allosteric regulation. Specifically, our findings reveal that the binding of one molecule of Sos1 to the nSH3 domain allosterically induces a conformational change within Grb2 such that the loading of a second molecule of Sos1 onto the cSH3 domain is blocked and, in so doing, allows Gab1 access to the cSH3 domain in an exclusively non-competitive manner to generate the Sos1-Grb2-Gab1 ternary signaling complex. PMID:20005866

  12. Long-range allosteric signaling in red light–regulated diguanylyl cyclases

    PubMed Central

    Gourinchas, Geoffrey; Etzl, Stefan; Göbl, Christoph; Vide, Uršula; Madl, Tobias; Winkler, Andreas

    2017-01-01

    Nature has evolved an astonishingly modular architecture of covalently linked protein domains with diverse functionalities to enable complex cellular networks that are critical for cell survival. The coupling of sensory modules with enzymatic effectors allows direct allosteric regulation of cellular signaling molecules in response to diverse stimuli. We present molecular details of red light–sensing bacteriophytochromes linked to cyclic dimeric guanosine monophosphate–producing diguanylyl cyclases. Elucidation of the first crystal structure of a full-length phytochrome with its enzymatic effector, in combination with the characterization of light-induced changes in conformational dynamics, reveals how allosteric light regulation is fine-tuned by the architecture and composition of the coiled-coil sensor-effector linker and also the central helical spine. We anticipate that consideration of molecular principles of sensor-effector coupling, going beyond the length of the characteristic linker, and the appreciation of dynamically driven allostery will open up new directions for the design of novel red light–regulated optogenetic tools. PMID:28275738

  13. Agonistic aptamer to the insulin receptor leads to biased signaling and functional selectivity through allosteric modulation

    PubMed Central

    Yunn, Na-Oh; Koh, Ara; Han, Seungmin; Lim, Jong Hun; Park, Sehoon; Lee, Jiyoun; Kim, Eui; Jang, Sung Key; Berggren, Per-Olof; Ryu, Sung Ho

    2015-01-01

    Due to their high affinity and specificity, aptamers have been widely used as effective inhibitors in clinical applications. However, the ability to activate protein function through aptamer-protein interaction has not been well-elucidated. To investigate their potential as target-specific agonists, we used SELEX to generate aptamers to the insulin receptor (IR) and identified an agonistic aptamer named IR-A48 that specifically binds to IR, but not to IGF-1 receptor. Despite its capacity to stimulate IR autophosphorylation, similar to insulin, we found that IR-A48 not only binds to an allosteric site distinct from the insulin binding site, but also preferentially induces Y1150 phosphorylation in the IR kinase domain. Moreover, Y1150-biased phosphorylation induced by IR-A48 selectively activates specific signaling pathways downstream of IR. In contrast to insulin-mediated activation of IR, IR-A48 binding has little effect on the MAPK pathway and proliferation of cancer cells. Instead, AKT S473 phosphorylation is highly stimulated by IR-A48, resulting in increased glucose uptake both in vitro and in vivo. Here, we present IR-A48 as a biased agonist able to selectively induce the metabolic activity of IR through allosteric binding. Furthermore, our study also suggests that aptamers can be a promising tool for developing artificial biased agonists to targeted receptors. PMID:26245346

  14. Biased signalling and allosteric machines: new vistas and challenges for drug discovery.

    PubMed

    Kenakin, Terry P

    2012-03-01

    Seven transmembrane receptors (7TMRs) are nature's prototype allosteric proteins made to bind molecules at one location to subsequently change their shape to affect the binding of another molecule at another location. This paper attempts to describe the divergent 7TMR behaviours (i.e. third party allostery, receptor oligomerization, biased agonism) observed in pharmacology in terms of a homogeneous group of allosteric behaviours. By considering the bodies involved as a vector defined by a modulator, conduit and guest, these activities can all be described by a simple model of functional allostery made up of the Ehlert allosteric model and the Black/Leff operational model. It will be shown how this model yields parameters that can be used to characterize the activity of any ligand or protein producing effect through allosteric interaction with a 7TMR.

  15. Allosteric signalling in the outer membrane translocation domain of PapC usher

    PubMed Central

    Farabella, Irene; Pham, Thieng; Henderson, Nadine S; Geibel, Sebastian; Phan, Gilles; Thanassi, David G; Delcour, Anne H; Waksman, Gabriel; Topf, Maya

    2014-01-01

    PapC ushers are outer-membrane proteins enabling assembly and secretion of P pili in uropathogenic E. coli. Their translocation domain is a large β-barrel occluded by a plug domain, which is displaced to allow the translocation of pilus subunits across the membrane. Previous studies suggested that this gating mechanism is controlled by a β-hairpin and an α-helix. To investigate the role of these elements in allosteric signal communication, we developed a method combining evolutionary and molecular dynamics studies of the native translocation domain and mutants lacking the β-hairpin and/or the α-helix. Analysis of a hybrid residue interaction network suggests distinct regions (residue ‘communities’) within the translocation domain (especially around β12–β14) linking these elements, thereby modulating PapC gating. Antibiotic sensitivity and electrophysiology experiments on a set of alanine-substitution mutants confirmed functional roles for four of these communities. This study illuminates the gating mechanism of PapC ushers and its importance in maintaining outer-membrane permeability. DOI: http://dx.doi.org/10.7554/eLife.03532.001 PMID:25271373

  16. The condensed chromatin fiber: an allosteric chemo-mechanical machine for signal transduction and genome processing

    NASA Astrophysics Data System (ADS)

    Lesne, Annick; Bécavin, Christophe; Victor, Jean–Marc

    2012-02-01

    Allostery is a key concept of molecular biology which refers to the control of an enzyme activity by an effector molecule binding the enzyme at another site rather than the active site (allos = other in Greek). We revisit here allostery in the context of chromatin and argue that allosteric principles underlie and explain the functional architecture required for spacetime coordination of gene expression at all scales from DNA to the whole chromosome. We further suggest that this functional architecture is provided by the chromatin fiber itself. The structural, mechanical and topological features of the chromatin fiber endow chromosomes with a tunable signal transduction from specific (or nonspecific) effectors to specific (or nonspecific) active sites. Mechanical constraints can travel along the fiber all the better since the fiber is more compact and regular, which speaks in favor of the actual existence of the (so-called 30 nm) chromatin fiber. Chromatin fiber allostery reconciles both the physical and biochemical approaches of chromatin. We illustrate this view with two supporting specific examples. Moreover, from a methodological point of view, we suggest that the notion of chromatin fiber allostery is particularly relevant for systemic approaches. Finally we discuss the evolutionary power of allostery in the context of chromatin and its relation to modularity.

  17. The allosteric behavior of Fur mediates oxidative stress signal transduction in Helicobacter pylori.

    PubMed

    Pelliciari, Simone; Vannini, Andrea; Roncarati, Davide; Danielli, Alberto

    2015-01-01

    The microaerophilic gastric pathogen Helicobacter pylori is exposed to oxidative stress originating from the aerobic environment, the oxidative burst of phagocytes and the formation of reactive oxygen species, catalyzed by iron excess. Accordingly, the expression of genes involved in oxidative stress defense have been repeatedly linked to the ferric uptake regulator Fur. Moreover, mutations in the Fur protein affect the resistance to metronidazole, likely due to loss-of-function in the regulation of genes involved in redox control. Although many advances in the molecular understanding of HpFur function were made, little is known about the mechanisms that enable Fur to mediate the responses to oxidative stress. Here we show that iron-inducible, apo-Fur repressed genes, such as pfr and hydA, are induced shortly after oxidative stress, while their oxidative induction is lost in a fur knockout strain. On the contrary, holo-Fur repressed genes, such as frpB1 and fecA1, vary modestly in response to oxidative stress. This indicates that the oxidative stress signal specifically targets apo-Fur repressed genes, rather than impairing indiscriminately the regulatory function of Fur. Footprinting analyses showed that the oxidative signal strongly impairs the binding affinity of Fur toward apo-operators, while the binding toward holo-operators is less affected. Further evidence is presented that a reduced state of Fur is needed to maintain apo-repression, while oxidative conditions shift the preferred binding architecture of Fur toward the holo-operator binding conformation, even in the absence of iron. Together the results demonstrate that the allosteric regulation of Fur enables transduction of oxidative stress signals in H. pylori, supporting the concept that apo-Fur repressed genes can be considered oxidation inducible Fur regulatory targets. These findings may have important implications in the study of H. pylori treatment and resistance to antibiotics.

  18. Allosteric Modulation of Metabotropic Glutamate Receptors

    PubMed Central

    Sheffler, Douglas J.; Gregory, Karen J.; Rook, Jerri M.; Conn, P. Jeffrey

    2013-01-01

    The development of receptor subtype-selective ligands by targeting allosteric sites of G protein-coupled receptors (GPCRs) has proven highly successful in recent years. One GPCR family that has greatly benefited from this approach is the metabotropic glutamate receptors (mGlus). These family C GPCRs participate in the neuromodulatory actions of glutamate throughout the CNS, where they play a number of key roles in regulating synaptic transmission and neuronal excitability. A large number of mGlu subtype-selective allosteric modulators have been identified, the majority of which are thought to bind within the transmembrane regions of the receptor. These modulators can either enhance or inhibit mGlu functional responses and, together with mGlu knockout mice, have furthered the establishment of the physiologic roles of many mGlu subtypes. Numerous pharmacological and receptor mutagenesis studies have been aimed at providing a greater mechanistic understanding of the interaction of mGlu allosteric modulators with the receptor, which have revealed evidence for common allosteric binding sites across multiple mGlu subtypes and the presence for multiple allosteric sites within a single mGlu subtype. Recent data have also revealed that mGlu allosteric modulators can display functional selectivity toward particular signal transduction cascades downstream of an individual mGlu subtype. Studies continue to validate the therapeutic utility of mGlu allosteric modulators as a potential therapeutic approach for a number of disorders including anxiety, schizophrenia, Parkinson’s disease, and Fragile X syndrome. PMID:21907906

  19. Differential pathway coupling efficiency of the activated insulin receptor drives signaling selectivity by XMetA, an allosteric partial agonist antibody

    Technology Transfer Automated Retrieval System (TEKTRAN)

    XMetA, an anti-insulin receptor (IR) monoclonal antibody, is an allosteric partial agonist of the IR. We have previously reported that XMetA activates the “metabolic-biased” Akt kinase signaling pathway while having little or no effect on the “mitogenic” MAPK signaling pathwayof ERK 1/2. To inves...

  20. Ligand-biased and probe-dependent modulation of chemokine receptor CXCR3 signaling by negative allosteric modulators.

    PubMed

    Bernat, Viachaslau; Brox, Regine; Heinrich, Markus R; Auberson, Yves P; Tschammer, Nuska

    2015-03-01

    Over the last decade, functional selectivity (or ligand bias) has evolved from being a peculiar phenomenon to being recognized as an essential feature of synthetic ligands that target G protein-coupled receptors (GPCRs). The CXC chemokine receptor 3 (CXCR3) is an outstanding platform to study various aspects of biased signaling, because nature itself uses functional selectivity to manipulate receptor signaling. At the same time, CXCR3 is an attractive therapeutic target in the treatment of autoimmune diseases and cancer. Herein we report the discovery of an 8-azaquinazolinone derivative (N-{1-[3-(4-ethoxyphenyl)-4-oxo-3,4-dihydropyrido[2,3-d]pyrimidin-2-yl]ethyl}-4-(4-fluorobutoxy)-N-[(1-methylpiperidin-4-yl)methyl]butanamide, 1 b) that can inhibit CXC chemokine 11 (CXCL11)-dependent G protein activation over β-arrestin recruitment with 187-fold selectivity. This compound also demonstrates probe-dependent activity, that is, it inhibits CXCL11- over CXCL10-mediated G protein activation with 12-fold selectivity. Together with a previously reported biased negative allosteric modulator from our group, the present study provides additional information on the molecular requirements for allosteric modulation of CXCR3.

  1. Mapping Cannabinoid 1 Receptor Allosteric Site(s): Critical Molecular Determinant and Signaling Profile of GAT100, a Novel, Potent, and Irreversibly Binding Probe.

    PubMed

    Laprairie, Robert B; Kulkarni, Abhijit R; Kulkarni, Pushkar M; Hurst, Dow P; Lynch, Diane; Reggio, Patricia H; Janero, David R; Pertwee, Roger G; Stevenson, Lesley A; Kelly, Melanie E M; Denovan-Wright, Eileen M; Thakur, Ganesh A

    2016-06-15

    One of the most abundant G-protein coupled receptors (GPCRs) in brain, the cannabinoid 1 receptor (CB1R), is a tractable therapeutic target for treating diverse psychobehavioral and somatic disorders. Adverse on-target effects associated with small-molecule CB1R orthosteric agonists and inverse agonists/antagonists have plagued their translational potential. Allosteric CB1R modulators offer a potentially safer modality through which CB1R signaling may be directed for therapeutic benefit. Rational design of candidate, druglike CB1R allosteric modulators requires greater understanding of the architecture of the CB1R allosteric endodomain(s) and the capacity of CB1R allosteric ligands to tune the receptor's information output. We have recently reported the synthesis of a focused library of rationally designed, covalent analogues of Org27569 and PSNCBAM-1, two prototypic CB1R negative allosteric modulators (NAMs). Among the novel, pharmacologically active CB1R NAMs reported, the isothiocyanate GAT100 emerged as the lead by virtue of its exceptional potency in the [(35)S]GTPγS and β-arrestin signaling assays and its ability to label CB1R as a covalent allosteric probe with significantly reduced inverse agonism in the [(35)S]GTPγS assay as compared to Org27569. We report here a comprehensive functional profiling of GAT100 across an array of important downstream cell-signaling pathways and analysis of its potential orthosteric probe-dependence and signaling bias. The results demonstrate that GAT100 is a NAM of the orthosteric CB1R agonist CP55,940 and the endocannabinoids 2-arachidonoylglycerol and anandamide for β-arrestin1 recruitment, PLCβ3 and ERK1/2 phosphorylation, cAMP accumulation, and CB1R internalization in HEK293A cells overexpressing CB1R and in Neuro2a and STHdh(Q7/Q7) cells endogenously expressing CB1R. Distinctively, GAT100 was a more potent and efficacious CB1R NAM than Org27569 and PSNCBAM-1 in all signaling assays and did not exhibit the inverse

  2. Michaelis-Menten at 100 and allosterism at 50: driving molecular motors in a hailstorm with noisy ATPase engines and allosteric transmission.

    PubMed

    Chowdhury, Debashish

    2014-01-01

    Cytoskeletal motor proteins move on filamentous tracks by converting input chemical energy that they derive by catalyzing the hydrolysis of ATP. The ATPase site is the analogue of an engine and hydrolysis of ATP is the analogue of burning of chemical fuel. Moreover, the functional role of a segment of the motor is analogous to that of the transmission system of an automobile, which consists of a shaft, gear, clutch, etc. The operation of the engine is intrinsically 'noisy' and the motor faces a molecular 'hailstorm' in the aqueous medium. In this commemorative review, we celebrate the centenary of Michaelis and Menten's landmark paper of 1913 and the golden jubilee of Monod and colleagues classic paper of 1963 by highlighting their relevance with respect to explaining the operational mechanisms of the engine and the transmission system, respectively, of cytoskeletal motors.

  3. Measurements on wireless transmission of ECG signals

    NASA Astrophysics Data System (ADS)

    Gabrielli, A.; Lax, I.

    2016-12-01

    The scope of this research is to design an electronic prototype, an operative system as a proof of concept, to transmit and receive biological parameters, in particular electrocardiogram signals, through dedicated wireless circuits. The apparatus features microelectronics chips that were developed for more general biomedical applications, here adapted to deal with cardiac signals. The paper mainly focuses on the electronic aspects, as in this study we do not face medical or clinical aspects of the system. The transmitter circuit uses a commercial instrumentation amplifier and the receiver has been equipped with wide-band amplifiers along with made-in-the-lab band-pass filters centered at the carrier. We have been able to mount the entire system prototype into a preliminary data acquisition chain that reads out the electrocardiogram signal. The prototype allows acquiring the waveform, converting it to a digital pattern and open the transmission through a series of high-frequency packets exploiting the Ultra Wide Band protocol. The sensor value is embedded in the transmission through the rate of the digital packets. In fact, these are sent wireless at a specific packet-frequency that depends on the sensor amplitude and are detected into a receiver circuit that recovers the information.

  4. Discovery of a novel allosteric modulator of 5-HT3 receptors: inhibition and potentiation of Cys-loop receptor signaling through a conserved transmembrane intersubunit site.

    PubMed

    Trattnig, Sarah M; Harpsøe, Kasper; Thygesen, Sarah B; Rahr, Louise M; Ahring, Philip K; Balle, Thomas; Jensen, Anders A

    2012-07-20

    The ligand-gated ion channels in the Cys-loop receptor superfamily mediate the effects of neurotransmitters acetylcholine, serotonin, GABA, and glycine. Cys-loop receptor signaling is susceptible to modulation by ligands acting through numerous allosteric sites. Here we report the discovery of a novel class of negative allosteric modulators of the 5-HT(3) receptors (5-HT(3)Rs). PU02 (6-[(1-naphthylmethyl)thio]-9H-purine) is a potent and selective antagonist displaying IC(50) values of ~1 μM at 5-HT(3)Rs and substantially lower activities at other Cys-loop receptors. In an elaborate mutagenesis study of the 5-HT(3)A receptor guided by a homology model, PU02 is demonstrated to act through a transmembrane intersubunit site situated in the upper three helical turns of TM2 and TM3 in the (+)-subunit and TM1 and TM2 in the (-)-subunit. The Ser(248), Leu(288), Ile(290), Thr(294), and Gly(306) residues are identified as important molecular determinants of PU02 activity with minor contributions from Ser(292) and Val(310), and we propose that the naphthalene group of PU02 docks into the hydrophobic cavity formed by these. Interestingly, specific mutations of Ser(248), Thr(294), and Gly(306) convert PU02 into a complex modulator, potentiating and inhibiting 5-HT-evoked signaling through these mutants at low and high concentrations, respectively. The PU02 binding site in the 5-HT(3)R corresponds to allosteric sites in anionic Cys-loop receptors, which emphasizes the uniform nature of the molecular events underlying signaling through the receptors. Moreover, the dramatic changes in the functional properties of PU02 induced by subtle changes in its binding site bear witness to the delicate structural discrimination between allosteric inhibition and potentiation of Cys-loop receptors.

  5. Dynamics of Mechanical Signal Transmission through Prestressed Stress Fibers

    PubMed Central

    Hwang, Yongyun; Barakat, Abdul I.

    2012-01-01

    Transmission of mechanical stimuli through the actin cytoskeleton has been proposed as a mechanism for rapid long-distance mechanotransduction in cells; however, a quantitative understanding of the dynamics of this transmission and the physical factors governing it remains lacking. Two key features of the actin cytoskeleton are its viscoelastic nature and the presence of prestress due to actomyosin motor activity. We develop a model of mechanical signal transmission through prestressed viscoelastic actin stress fibers that directly connect the cell surface to the nucleus. The analysis considers both temporally stationary and oscillatory mechanical signals and accounts for cytosolic drag on the stress fibers. To elucidate the physical parameters that govern mechanical signal transmission, we initially focus on the highly simplified case of a single stress fiber. The results demonstrate that the dynamics of mechanical signal transmission depend on whether the applied force leads to transverse or axial motion of the stress fiber. For transverse motion, mechanical signal transmission is dominated by prestress while fiber elasticity has a negligible effect. Conversely, signal transmission for axial motion is mediated uniquely by elasticity due to the absence of a prestress restoring force. Mechanical signal transmission is significantly delayed by stress fiber material viscosity, while cytosolic damping becomes important only for longer stress fibers. Only transverse motion yields the rapid and long-distance mechanical signal transmission dynamics observed experimentally. For simple networks of stress fibers, mechanical signals are transmitted rapidly to the nucleus when the fibers are oriented largely orthogonal to the applied force, whereas the presence of fibers parallel to the applied force slows down mechanical signal transmission significantly. The present results suggest that cytoskeletal prestress mediates rapid mechanical signal transmission and allows

  6. Dynamics of mechanical signal transmission through prestressed stress fibers.

    PubMed

    Hwang, Yongyun; Barakat, Abdul I

    2012-01-01

    Transmission of mechanical stimuli through the actin cytoskeleton has been proposed as a mechanism for rapid long-distance mechanotransduction in cells; however, a quantitative understanding of the dynamics of this transmission and the physical factors governing it remains lacking. Two key features of the actin cytoskeleton are its viscoelastic nature and the presence of prestress due to actomyosin motor activity. We develop a model of mechanical signal transmission through prestressed viscoelastic actin stress fibers that directly connect the cell surface to the nucleus. The analysis considers both temporally stationary and oscillatory mechanical signals and accounts for cytosolic drag on the stress fibers. To elucidate the physical parameters that govern mechanical signal transmission, we initially focus on the highly simplified case of a single stress fiber. The results demonstrate that the dynamics of mechanical signal transmission depend on whether the applied force leads to transverse or axial motion of the stress fiber. For transverse motion, mechanical signal transmission is dominated by prestress while fiber elasticity has a negligible effect. Conversely, signal transmission for axial motion is mediated uniquely by elasticity due to the absence of a prestress restoring force. Mechanical signal transmission is significantly delayed by stress fiber material viscosity, while cytosolic damping becomes important only for longer stress fibers. Only transverse motion yields the rapid and long-distance mechanical signal transmission dynamics observed experimentally. For simple networks of stress fibers, mechanical signals are transmitted rapidly to the nucleus when the fibers are oriented largely orthogonal to the applied force, whereas the presence of fibers parallel to the applied force slows down mechanical signal transmission significantly. The present results suggest that cytoskeletal prestress mediates rapid mechanical signal transmission and allows

  7. Transmission of isolated LVDS signal pulses at long distance

    NASA Astrophysics Data System (ADS)

    Formenti, F.; Scapparone, E.

    2010-03-01

    We discuss in this paper the challenge of transmitting isolated LVDS (Low Voltage Differential Signaling) pulses over long cables ( L> 40 m). A safe approach is developed paying special care in the choice of the components used at the different stages of the data transmission. In each step, starting from the signal generation, then facing the propagation through the cable and finally addressing the signal detection at the far end, we tried to preserve the signal shape, while reducing the sources of the signal attenuation and distortion. Two different approaches for a successful data transmission are discussed, both allowing a safe performance.

  8. Signal transmissibility in marginal granular materials

    NASA Astrophysics Data System (ADS)

    Pinson, Matthew B.; Witten, Thomas A.

    2016-12-01

    We examine the ‘transmissibility’ of a simulated two-dimensional pack of frictionless disks formed by confining dilute disks in a shrinking, periodic box to the point of mechanical stability. Two opposite boundaries are then removed, thus allowing a set of free motions. Small free displacements on one boundary then induce proportional displacements on the opposite boundary. Transmissibility is the ability to distinguish different perturbations by their distant responses. We assess transmissibility by successively identifying free orthonormal modes of motion that have the smallest distant responses. The last modes to be identified in this ‘pessimistic’ basis are the most transmissive. The transmitted amplitudes of these most transmissive modes fall off exponentially with mode number. Similar exponential falloff is seen in a simple elastic medium, though the responsible modes differ greatly in structure in the two systems. Thus the marginal pack’s transmissibility is qualitatively similar to that of a simple elastic medium. We compare our results with recent findings based on the projection of the space of free motion onto interior sites.

  9. Frequency Spreading in Underwater Acoustic Signal Transmission.

    DTIC Science & Technology

    1980-04-15

    acoustic signal transmitted and received underwater J-2 J.2 Signal spectrum computing block diagram. J-3 Chapter I. Frequency spreading 1.0 Introduction... transmitted frequency can be expected in the received signal [1] - [18]. This frequency spreading behavior is the result of the amplitude and phase...result of phase modulation of the transmitted sinusoid by the moving surface, and the separation between the spectral lines at the receiving point is

  10. Dynamic coupling and allosteric behavior in a non-allosteric protein†

    PubMed Central

    Clarkson, Michael W.; Gilmore, Steven A.; Edgell, Marshall H.; Lee, Andrew L.

    2008-01-01

    Long-range intraprotein interactions give rise to many important protein behaviors. Understanding how energy is transduced through protein structures to either transmit a signal or elicit conformational changes is therefore a current challenge in structural biology. In an effort to understand such linkages, multiple V→A mutations were made in the small globular protein eglin c. The physical responses, as mapped by NMR spin relaxation, residual dipolar couplings (RDCs), and scalar couplings, illustrate that the interior of this non-allosteric protein forms a dynamic network and that local perturbations are transmitted as dynamic and structural changes to distal sites as far as 16 Å away. Two basic types of propagation responses were observed: contiguous pathways of enhanced (attenuated) dynamics with no change in structure; and dispersed (non-contiguous) changes in methyl rotation rates that appear to result from subtle deformation of backbone structure. In addition, energy transmission is found to be unidirectional. In one mutant, an allosteric conformational change of a side chain is seen in the context of a pathway of propagated changes in ps-ns dynamics. The observation of so many long-range interactions in a small, rigid system lends experimental weight to the idea that all well-folded proteins inherently possess allosteric features [Gunasekaran et al. (2004) Proteins 57, 433−443], and that dynamics are a rich source of information for mapping and gaining mechanistic insight into communication pathways in individual proteins. PMID:16784220

  11. Allosteric Modulation of Chemoattractant Receptors

    PubMed Central

    Allegretti, Marcello; Cesta, Maria Candida; Locati, Massimo

    2016-01-01

    Chemoattractants control selective leukocyte homing via interactions with a dedicated family of related G protein-coupled receptor (GPCR). Emerging evidence indicates that the signaling activity of these receptors, as for other GPCR, is influenced by allosteric modulators, which interact with the receptor in a binding site distinct from the binding site of the agonist and modulate the receptor signaling activity in response to the orthosteric ligand. Allosteric modulators have a number of potential advantages over orthosteric agonists/antagonists as therapeutic agents and offer unprecedented opportunities to identify extremely selective drug leads. Here, we resume evidence of allosterism in the context of chemoattractant receptors, discussing in particular its functional impact on functional selectivity and probe/concentration dependence of orthosteric ligands activities. PMID:27199992

  12. Dependence effects in unique signal transmission

    SciTech Connect

    Cooper, J.A.

    1988-04-01

    ''Unique Signals'' are communicated from a source to a ''strong link'' safety device in a weapon by means of one or more digital communication channels. The probability that the expected unique signal pattern could be generated accidentally (e.g., due to an abnormal environment) would be an important measure. A probabilistic assessment of this likelihood is deceptive, because it depends on characteristics of the other traffic on the communication channel. One such characteristic that is frequently neglected in analysis is dependence. This report gives a mathematical model for dependence; cites some of the ways in which dependence can increase the likelihood of inadvertent unique signal pattern generation; and suggests that communicating each unique signal ''event'' at the highest level of protocol in the communication system minimizes dependence effects. 3 refs., 4 figs.

  13. Mechanisms of cytoskeleton-mediated mechanical signal transmission in cells

    PubMed Central

    Hwang, Yongyun; Gouget, Cecile L.M.; Barakat, Abdul I.

    2012-01-01

    Recent experiments have demonstrated very rapid long-distance transmission of mechanical forces within cells. Because the speed of this transmission greatly exceeds that of reaction-diffusion signaling, it has been conjectured that it occurs via the propagation of elastic waves through the actin stress fiber network. To explore the plausibility of this conjecture, we recently developed a model of small amplitude stress fiber deformations in prestressed viscoelastic stress fibers subjected to external forces. The model results demonstrated that rapid mechanical signal transmission is only possible when the external force is applied orthogonal to the stress fiber axis and that the dynamics of this transmission are governed by a balance between the prestress in the stress fiber and the stress fiber's material viscosity. The present study, which is a follow-up on our previous model, uses dimensional analysis to: (1) further evaluate the plausibility of the elastic wave conjecture and (2) obtain insight into mechanical signal transmission dynamics in simple stress fiber networks. We show that the elastic wave scenario is likely not the mechanism of rapid mechanical signal transmission in actin stress fibers due to the highly viscoelastic character of these fibers. Our analysis also demonstrates that the time constant characterizing mechanical stimulus transmission is strongly dependent on the topology of the stress fiber network, implying that network organization plays an important role in determining the dynamics of cellular responsiveness to mechanical stimulation. PMID:23336020

  14. Mechanisms of cytoskeleton-mediated mechanical signal transmission in cells.

    PubMed

    Hwang, Yongyun; Gouget, Cecile L M; Barakat, Abdul I

    2012-11-01

    Recent experiments have demonstrated very rapid long-distance transmission of mechanical forces within cells. Because the speed of this transmission greatly exceeds that of reaction-diffusion signaling, it has been conjectured that it occurs via the propagation of elastic waves through the actin stress fiber network. To explore the plausibility of this conjecture, we recently developed a model of small amplitude stress fiber deformations in prestressed viscoelastic stress fibers subjected to external forces. The model results demonstrated that rapid mechanical signal transmission is only possible when the external force is applied orthogonal to the stress fiber axis and that the dynamics of this transmission are governed by a balance between the prestress in the stress fiber and the stress fiber's material viscosity. The present study, which is a follow-up on our previous model, uses dimensional analysis to: (1) further evaluate the plausibility of the elastic wave conjecture and (2) obtain insight into mechanical signal transmission dynamics in simple stress fiber networks. We show that the elastic wave scenario is likely not the mechanism of rapid mechanical signal transmission in actin stress fibers due to the highly viscoelastic character of these fibers. Our analysis also demonstrates that the time constant characterizing mechanical stimulus transmission is strongly dependent on the topology of the stress fiber network, implying that network organization plays an important role in determining the dynamics of cellular responsiveness to mechanical stimulation.

  15. High bandwidth magnetically isolated signal transmission circuit

    NASA Technical Reports Server (NTRS)

    Repp, John Donald (Inventor)

    2005-01-01

    Many current electronic systems incorporate expensive or sensitive electrical components. Because electrical energy is often generated or transmitted at high voltages, the power supplies to these electronic systems must be carefully designed. Power supply design must ensure that the electrical system being supplied with power is not exposed to excessive voltages or currents. In order to isolate power supplies from electrical equipment, many methods have been employed. These methods typically involve control systems or signal transfer methods. However, these methods are not always suitable because of their drawbacks. The present invention relates to transmitting information across an interface. More specifically, the present invention provides an apparatus for transmitting both AC and DC information across a high bandwidth magnetic interface with low distortion.

  16. Exploring Molecular Mechanisms of Paradoxical Activation in the BRAF Kinase Dimers: Atomistic Simulations of Conformational Dynamics and Modeling of Allosteric Communication Networks and Signaling Pathways

    PubMed Central

    Tse, Amanda; Verkhivker, Gennady M.

    2016-01-01

    The recent studies have revealed that most BRAF inhibitors can paradoxically induce kinase activation by promoting dimerization and enzyme transactivation. Despite rapidly growing number of structural and functional studies about the BRAF dimer complexes, the molecular basis of paradoxical activation phenomenon is poorly understood and remains largely hypothetical. In this work, we have explored the relationships between inhibitor binding, protein dynamics and allosteric signaling in the BRAF dimers using a network-centric approach. Using this theoretical framework, we have combined molecular dynamics simulations with coevolutionary analysis and modeling of the residue interaction networks to determine molecular determinants of paradoxical activation. We have investigated functional effects produced by paradox inducer inhibitors PLX4720, Dabrafenib, Vemurafenib and a paradox breaker inhibitor PLX7904. Functional dynamics and binding free energy analyses of the BRAF dimer complexes have suggested that negative cooperativity effect and dimer-promoting potential of the inhibitors could be important drivers of paradoxical activation. We have introduced a protein structure network model in which coevolutionary residue dependencies and dynamic maps of residue correlations are integrated in the construction and analysis of the residue interaction networks. The results have shown that coevolutionary residues in the BRAF structures could assemble into independent structural modules and form a global interaction network that may promote dimerization. We have also found that BRAF inhibitors could modulate centrality and communication propensities of global mediating centers in the residue interaction networks. By simulating allosteric communication pathways in the BRAF structures, we have determined that paradox inducer and breaker inhibitors may activate specific signaling routes that correlate with the extent of paradoxical activation. While paradox inducer inhibitors may

  17. Oscillation-Induced Signal Transmission and Gating in Neural Circuits

    PubMed Central

    Jahnke, Sven; Memmesheimer, Raoul-Martin; Timme, Marc

    2014-01-01

    Reliable signal transmission constitutes a key requirement for neural circuit function. The propagation of synchronous pulse packets through recurrent circuits is hypothesized to be one robust form of signal transmission and has been extensively studied in computational and theoretical works. Yet, although external or internally generated oscillations are ubiquitous across neural systems, their influence on such signal propagation is unclear. Here we systematically investigate the impact of oscillations on propagating synchrony. We find that for standard, additive couplings and a net excitatory effect of oscillations, robust propagation of synchrony is enabled in less prominent feed-forward structures than in systems without oscillations. In the presence of non-additive coupling (as mediated by fast dendritic spikes), even balanced oscillatory inputs may enable robust propagation. Here, emerging resonances create complex locking patterns between oscillations and spike synchrony. Interestingly, these resonances make the circuits capable of selecting specific pathways for signal transmission. Oscillations may thus promote reliable transmission and, in co-action with dendritic nonlinearities, provide a mechanism for information processing by selectively gating and routing of signals. Our results are of particular interest for the interpretation of sharp wave/ripple complexes in the hippocampus, where previously learned spike patterns are replayed in conjunction with global high-frequency oscillations. We suggest that the oscillations may serve to stabilize the replay. PMID:25503492

  18. Transmission of RF Signals Over Optical Fiber for Avionics Applications

    NASA Technical Reports Server (NTRS)

    Slaveski, Filip; Sluss, James, Jr.; Atiquzzaman, Mohammed; Hung, Nguyen; Ngo, Duc

    2002-01-01

    During flight, aircraft avionics transmit and receive RF signals to/from antennas over coaxial cables. As the density and complexity of onboard avionics increases, the electromagnetic interference (EM) environment degrades proportionately, leading to decreasing signal-to-noise ratios (SNRs) and potential safety concerns. The coaxial cables are inherently lossy, limiting the RF signal bandwidth while adding considerable weight. To overcome these limitations, we have investigated a fiber optic communications link for aircraft that utilizes wavelength division multiplexing (WDM) to support the simultaneous transmission of multiple signals (including RF) over a single optical fiber. Optical fiber has many advantages over coaxial cable, particularly lower loss, greater bandwidth, and immunity to EM. In this paper, we demonstrate that WDM can be successfully used to transmit multiple RF signals over a single optical fiber with no appreciable signal degradation. We investigate the transmission of FM and AM analog modulated signals, as well as FSK digital modulated signals, over a fiber optic link (FOL) employing WDM. We present measurements of power loss, delay, SNR, carrier-to-noise ratio (CNR), total harmonic distortion (THD), and bit error rate (BER). Our experimental results indicate that WDM is a fiber optic technology suitable for avionics applications.

  19. Hub-activated signal transmission in complex networks

    NASA Astrophysics Data System (ADS)

    Jahnke, Sven; Memmesheimer, Raoul-Martin; Timme, Marc

    2014-03-01

    A wide range of networked systems exhibit highly connected nodes (hubs) as prominent structural elements. The functional roles of hubs in the collective nonlinear dynamics of many such networks, however, are not well understood. Here, we propose that hubs in neural circuits may activate local signal transmission along sequences of specific subnetworks. Intriguingly, in contrast to previous suggestions of the functional roles of hubs, here, not the hubs themselves, but nonhub subnetworks transfer the signals. The core mechanism relies on hubs and nonhubs providing activating feedback to each other. It may, thus, induce the propagation of specific pulse and rate signals in neuronal and other communication networks.

  20. Regulation of NMDA-receptor synaptic transmission by Wnt signaling

    PubMed Central

    Cerpa, Waldo; Gambrill, Abigail; Inestrosa, Nibaldo C.; Barria, Andres

    2011-01-01

    Wnt ligands are secreted glycoproteins controlling gene expression and cytoskeleton reorganization involved in embryonic development of the nervous system. However, their role in later stages of brain development, particularly in the regulation of established synaptic connections is not known. We found that Wnt-5a acutely and specifically up-regulates synaptic NMDAR currents in rat hippocampal slices facilitating induction of LTP, a cellular model of learning and memory. This effect requires an increase in postsynaptic Ca2+ and activation of non-canonical downstream effectors of the Wnt signaling pathway. In contrast, Wnt-7a, an activator of the canonical Wnt signaling pathway, has no effect on NMDAR mediated synaptic transmission. Moreover, endogenous Wnt ligands are necessary to maintain basal NMDAR synaptic transmission adjusting the threshold for synaptic potentiation. This novel role for Wnt ligands provides a mechanism for Wnt signaling to acutely modulate synaptic plasticity and brain function in later stages of development and in the mature organism. PMID:21715611

  1. 29 CFR 1926.1420 - Signals-radio, telephone or other electronic transmission of signals.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 29 Labor 8 2013-07-01 2013-07-01 false Signals-radio, telephone or other electronic transmission of signals. 1926.1420 Section 1926.1420 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL... CONSTRUCTION Cranes and Derricks in Construction § 1926.1420 Signals—radio, telephone or other...

  2. 29 CFR 1926.1420 - Signals-radio, telephone or other electronic transmission of signals.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 29 Labor 8 2011-07-01 2011-07-01 false Signals-radio, telephone or other electronic transmission of signals. 1926.1420 Section 1926.1420 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL... CONSTRUCTION Cranes and Derricks in Construction § 1926.1420 Signals—radio, telephone or other...

  3. 29 CFR 1926.1420 - Signals-radio, telephone or other electronic transmission of signals.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 29 Labor 8 2012-07-01 2012-07-01 false Signals-radio, telephone or other electronic transmission of signals. 1926.1420 Section 1926.1420 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL... CONSTRUCTION Cranes and Derricks in Construction § 1926.1420 Signals—radio, telephone or other...

  4. 29 CFR 1926.1420 - Signals-radio, telephone or other electronic transmission of signals.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 29 Labor 8 2014-07-01 2014-07-01 false Signals-radio, telephone or other electronic transmission of signals. 1926.1420 Section 1926.1420 Labor Regulations Relating to Labor (Continued) OCCUPATIONAL... CONSTRUCTION Cranes and Derricks in Construction § 1926.1420 Signals—radio, telephone or other...

  5. Calculate bit error rate for digital radio signal transmission

    NASA Astrophysics Data System (ADS)

    Sandberg, Jorgen

    1987-06-01

    A method for estimating symbol error rate caused by imperfect transmission channels is proposed. The method relates the symbol error rate to peak-to-peak amplitude and phase ripple, maximum gain slope, and maximum group delay distortion. The performance degradation of QPSK, offset QPSK (OQPSK), M-ary PSK (MSK) signals transmitted over a wideband channel, exhibiting either sinusoidal amplitude or phase ripples are evaluated using the proposed method. The transmission channel model, which is a single filter with transfer characteristics, for modeling the frequency of response of a system is described. Consideration is given to signal detection and system degradation. The calculations reveal that the QPSK modulated carrier degrades less then the OQPSK and MSK carriers for peak-to-peak amplitude ripple values less than 6 dB and peak-to-peak phase ripple values less than 45 deg.

  6. Biophoton signal transmission and processing in the brain.

    PubMed

    Tang, Rendong; Dai, Jiapei

    2014-10-05

    The transmission and processing of neural information in the nervous system plays a key role in neural functions. It is well accepted that neural communication is mediated by bioelectricity and chemical molecules via the processes called bioelectrical and chemical transmission, respectively. Indeed, the traditional theories seem to give valuable explanations for the basic functions of the nervous system, but difficult to construct general accepted concepts or principles to provide reasonable explanations of higher brain functions and mental activities, such as perception, learning and memory, emotion and consciousness. Therefore, many unanswered questions and debates over the neural encoding and mechanisms of neuronal networks remain. Cell to cell communication by biophotons, also called ultra-weak photon emissions, has been demonstrated in several plants, bacteria and certain animal cells. Recently, both experimental evidence and theoretical speculation have suggested that biophotons may play a potential role in neural signal transmission and processing, contributing to the understanding of the high functions of nervous system. In this paper, we review the relevant experimental findings and discuss the possible underlying mechanisms of biophoton signal transmission and processing in the nervous system.

  7. A 16-channel flex circuit for cryogenic microwave signal transmission

    NASA Astrophysics Data System (ADS)

    McGarey, Patrick; Mani, Hamdi; Wheeler, Caleb; Groppi, Christopher

    2014-07-01

    Heterodyne focal plane arrays used in the terahertz (THz) regime currently require a discrete set of rigid coaxial cables for the transmission of individual intermediate frequency (IF) signals. Consequently, the size of an array is limited to ~10s of pixels due to limited physical space and the complexity of assembly. In order to achieve an array with ~1000 pixels or greater, new interconnections must be developed capable of carrying multiple IF signals on a single carrier which is flexible, robust to noise, and terminated with a high density RF connector. As an intermediate step to the development of a ~1000 pixel heterodyne focal plane array, the Kilopixel Array Pathfinder Project (KAPPa) has developed a 16 channel IF flex circuit. Initially, design simulations were performed to evaluate various means of high-frequency (1~10 GHz) signal transmission, including microstrip, stripline and coplanar waveguides. The method allowing for the closest signal spacing and greatest resistance to radio frequency interference (RFI) was determined to be stripline. Designs were considered where stripline transitioned to microstrip in order to terminate the signal. As microstrip transmission lines are sensitive to RFI, a design featuring just stripline was evaluated. In both the stripline-to-microstrip and stripline-only designs, a three-layer copper-coated polyimide substrate was used. Signal transitions were accomplished by a signal carrying "hot" via passing through a series of three conductive pads, similar to work by Leib et al. (2010). The transition design essentially mimics a coaxial line, where the radial distance between the pads and the ground plane is optimized in order to achieve desired impedances. In simulation, 50 Ohm impedances were achieved throughout, with crosstalk and return loss limited to -30dB. Terminations are made via an array of Corning Gilbert G3PO blind mate connectors, which are small enough to match the 6mm pixel pitch of the KAPPa focal plane unit

  8. Differential Modulation of Functional Dynamics and Allosteric Interactions in the Hsp90-Cochaperone Complexes with p23 and Aha1: A Computational Study

    PubMed Central

    Blacklock, Kristin; Verkhivker, Gennady M.

    2013-01-01

    Allosteric interactions of the molecular chaperone Hsp90 with a large cohort of cochaperones and client proteins allow for molecular communication and event coupling in signal transduction networks. The integration of cochaperones into the Hsp90 system is driven by the regulatory mechanisms that modulate the progression of the ATPase cycle and control the recruitment of the Hsp90 clientele. In this work, we report the results of computational modeling of allosteric regulation in the Hsp90 complexes with the cochaperones p23 and Aha1. By integrating protein docking, biophysical simulations, modeling of allosteric communications, protein structure network analysis and the energy landscape theory we have investigated dynamics and stability of the Hsp90-p23 and Hsp90-Aha1 interactions in direct comparison with the extensive body of structural and functional experiments. The results have revealed that functional dynamics and allosteric interactions of Hsp90 can be selectively modulated by these cochaperones via specific targeting of the regulatory hinge regions that could restrict collective motions and stabilize specific chaperone conformations. The protein structure network parameters have quantified the effects of cochaperones on conformational stability of the Hsp90 complexes and identified dynamically stable communities of residues that can contribute to the strengthening of allosteric interactions. According to our results, p23-mediated changes in the Hsp90 interactions may provide “molecular brakes” that could slow down an efficient transmission of the inter-domain allosteric signals, consistent with the functional role of p23 in partially inhibiting the ATPase cycle. Unlike p23, Aha1-mediated acceleration of the Hsp90-ATPase cycle may be achieved via modulation of the equilibrium motions that facilitate allosteric changes favoring a closed dimerized form of Hsp90. The results of our study have shown that Aha1 and p23 can modulate the Hsp90-ATPase activity

  9. Markov propagation of allosteric effects in biomolecular systems: application to GroEL–GroES

    PubMed Central

    Chennubhotla, Chakra; Bahar, Ivet

    2006-01-01

    We introduce a novel approach for elucidating the potential pathways of allosteric communication in biomolecular systems. The methodology, based on Markov propagation of ‘information' across the structure, permits us to partition the network of interactions into soft clusters distinguished by their coherent stochastics. Probabilistic participation of residues in these clusters defines the communication patterns inherent to the network architecture. Application to bacterial chaperonin complex GroEL–GroES, an allostery-driven structure, identifies residues engaged in intra- and inter-subunit communication, including those acting as hubs and messengers. A number of residues are distinguished by their high potentials to transmit allosteric signals, including Pro33 and Thr90 at the nucleotide-binding site and Glu461 and Arg197 mediating inter- and intra-ring communication, respectively. We propose two most likely pathways of signal transmission, between nucleotide- and GroES-binding sites across the cis and trans rings, which involve several conserved residues. A striking observation is the opposite direction of information flow within cis and trans rings, consistent with negative inter-ring cooperativity. Comparison with collective modes deduced from normal mode analysis reveals the propensity of global hinge regions to act as messengers in the transmission of allosteric signals. PMID:16820777

  10. Signal processing for an optical wide band data transmission system

    NASA Astrophysics Data System (ADS)

    Nakamura, M.; Leskovar, B.; Turko, B. T.

    1987-07-01

    The signal processing for an optical wide band transmission system using gallium arsenide (GaAs) digital integrated circuits and optical fibers has been investigated. Multiplexing, coding, synchronization, demultiplexing, and error checking at 780 Mbit/s data rates are described. Data storage in memory for linking to a computer is also considered. The design uses available GaAs and silicon components. The reliability of GaAs components is discussed as well as the layout and thermal considerations required for a high speed system.

  11. Mechanical noise enhances signal transmission in the bullfrog sacculus.

    PubMed

    Indresano, Andrew A; Frank, Jonathan E; Middleton, Pameia; Jaramillo, Fernán

    2003-09-01

    Noise has been commonly thought to degrade the performance of sensory systems. However, it is now clear that the detection and transmission of weak signals in sensory systems can be enhanced by noise via stochastic resonance (SR). In hair cells, the quality of mechanoelectrical transduction is enhanced up to twofold by nanometer level mechanical noise acting on the hair bundle. We wanted to know whether these gains could be preserved, perhaps even enhanced, as information flows across hair cell synapses, and into the stream of action potentials that in the frog conveys acoustic information to the central nervous system. To approach this question, we studied the effects of noise on the signal-to-noise ratio (SNR) of the 8th nerve's response to small mechanical stimuli directly applied to the amphibian sacculus. We found that approximately 2.5 nm of mechanical noise enhanced the response of the saccular nerve up to fourfold, suggesting that the positive effects of low-amplitude mechanical noise result in improved transmission of acoustic information.

  12. Transmission of multiplexed video signals in multimode optical fiber systems

    NASA Technical Reports Server (NTRS)

    White, Preston, III

    1988-01-01

    Kennedy Space Center has the need for economical transmission of two multiplexed video signals along multimode fiberoptic systems. These systems must span unusual distances and must meet RS-250B short-haul standards after reception. Bandwidth is a major problem and studies of the installed fibers, available LEDs and PINFETs led to the choice of 100 MHz as the upper limit for the system bandwidth. Optical multiplexing and digital transmission were deemed inappropriate. Three electrical multiplexing schemes were chosen for further study. Each of the multiplexing schemes included an FM stage to help meet the stringent S/N specification. Both FM and AM frequency division multiplexing methods were investigated theoretically and these results were validated with laboratory tests. The novel application of quadrature amplitude multiplexing was also considered. Frequency division multiplexing of two wideband FM video signal appears the most promising scheme although this application requires high power highly linear LED transmitters. Futher studies are necessary to determine if LEDs of appropriate quality exist and to better quantify performance of QAM in this application.

  13. Transmission of multiplexed video signals in multimode optical fiber systems

    NASA Astrophysics Data System (ADS)

    White, Preston, III

    1988-10-01

    Kennedy Space Center has the need for economical transmission of two multiplexed video signals along multimode fiberoptic systems. These systems must span unusual distances and must meet RS-250B short-haul standards after reception. Bandwidth is a major problem and studies of the installed fibers, available LEDs and PINFETs led to the choice of 100 MHz as the upper limit for the system bandwidth. Optical multiplexing and digital transmission were deemed inappropriate. Three electrical multiplexing schemes were chosen for further study. Each of the multiplexing schemes included an FM stage to help meet the stringent S/N specification. Both FM and AM frequency division multiplexing methods were investigated theoretically and these results were validated with laboratory tests. The novel application of quadrature amplitude multiplexing was also considered. Frequency division multiplexing of two wideband FM video signal appears the most promising scheme although this application requires high power highly linear LED transmitters. Futher studies are necessary to determine if LEDs of appropriate quality exist and to better quantify performance of QAM in this application.

  14. Mechanistic insight into the conserved allosteric regulation of periplasmic proteolysis by the signaling molecule cyclic-di-GMP

    PubMed Central

    Chatterjee, Debashree; Cooley, Richard B; Boyd, Chelsea D; Mehl, Ryan A; O'Toole, George A; Sondermann, Holger

    2014-01-01

    Stable surface adhesion of cells is one of the early pivotal steps in bacterial biofilm formation, a prevalent adaptation strategy in response to changing environments. In Pseudomonas fluorescens, this process is regulated by the Lap system and the second messenger cyclic-di-GMP. High cytoplasmic levels of cyclic-di-GMP activate the transmembrane receptor LapD that in turn recruits the periplasmic protease LapG, preventing it from cleaving a cell surface-bound adhesin, thereby promoting cell adhesion. In this study, we elucidate the molecular basis of LapG regulation by LapD and reveal a remarkably sensitive switching mechanism that is controlled by LapD's HAMP domain. LapD appears to act as a coincidence detector, whereby a weak interaction of LapG with LapD transmits a transient outside-in signal that is reinforced only when cyclic-di-GMP levels increase. Given the conservation of key elements of this receptor system in many bacterial species, the results are broadly relevant for cyclic-di-GMP- and HAMP domain-regulated transmembrane signaling. DOI: http://dx.doi.org/10.7554/eLife.03650.001 PMID:25182848

  15. Allosteric Transmission along a Loosely Structured Backbone Allows a Cardiac Troponin C Mutant to Function with Only One Ca(2+) Ion.

    PubMed

    Marques, Mayra de A; Pinto, Jose Renato; Moraes, Adolfo H; Iqbal, Anwar; de Magalhães, Mariana T Q; Monteiro, Jamila; Pedrote, Murilo M; Sorenson, Martha M; Silva, Jerson L; de Oliveira, Guilherme A P

    2017-02-10

    Hypertrophic cardiomyopathy (HCM) is one of the most common cardiomyopathies and a major cause of sudden death in young athletes. The Ca(2+) sensor of the sarcomere, cardiac troponin C (cTnC), plays an important role in regulating muscle contraction. Although several cardiomyopathy-causing mutations have been identified in cTnC, the limited information about their structural defects has been mapped to the HCM phenotype. Here, we used high-resolution electron-spray ionization mass spectrometry (ESI-MS), Carr-Purcell-Meiboom-Gill relaxation dispersion (CPMG-RD), and affinity measurements of cTnC for the thin filament in reconstituted papillary muscles to provide evidence of an allosteric mechanism in mutant cTnC that may play a role to the HCM phenotype. We showed that the D145E mutation leads to altered dynamics on a μs-ms time scale and deactivates both of the divalent cation-binding sites of the cTnC C-domain. CPMG-RD captured a low populated protein-folding conformation triggered by the Glu-145 replacement of Asp. Paradoxically, although D145E C-domain was unable to bind Ca(2+), these changes along its backbone allowed it to attach more firmly to thin filaments than the wild-type isoform, providing evidence for an allosteric response of the Ca(2+)-binding site II in the N-domain. Our findings explain how the effects of an HCM mutation in the C-domain reflect up into the N-domain to cause an increase of Ca(2+) affinity in site II, thus opening up new insights into the HCM phenotype.

  16. Power and signal transmission for mobile teleoperated systems

    SciTech Connect

    Morris, A.C. Jr.; Hamel, W.R.

    1985-01-01

    Appropriate means must be furnished for supplying power and for sending controlling commands to mobile teleoperated systems. Because a sizable number of possibilities are available for such applications, methods used in designing both the power and communications systems built into mobile vehicles that serve in radiological emergencies must be carefully selected. This paper describes a number of umbilical, on-board, and wireless systems used in tranmitting power that are available for mobile teleoperator services. The pros and cons of selecting appropriate methods from a list of possible communication systems (wired, fiber optic, and radio frequency) are also examined. Moreover, hybrid systems combining wireless power transmissions with command-information signals are also possible.

  17. Computational Analysis of Residue Interaction Networks and Coevolutionary Relationships in the Hsp70 Chaperones: A Community-Hopping Model of Allosteric Regulation and Communication

    PubMed Central

    Stetz, Gabrielle; Verkhivker, Gennady M.

    2017-01-01

    introduced a community-hopping model of allosteric communication. Atomistic reconstruction of signaling pathways in the DnaK structures captured a direction-specific mechanism and molecular details of signal transmission that are fully consistent with the mutagenesis experiments. The results of our study reconciled structural and functional experiments from a network-centric perspective by showing that global properties of the residue interaction networks and coevolutionary signatures may be linked with specificity and diversity of allosteric regulation mechanisms. PMID:28095400

  18. Computational Modeling of Allosteric Regulation in the Hsp90 Chaperones: A Statistical Ensemble Analysis of Protein Structure Networks and Allosteric Communications

    PubMed Central

    Blacklock, Kristin; Verkhivker, Gennady M.

    2014-01-01

    A fundamental role of the Hsp90 chaperone in regulating functional activity of diverse protein clients is essential for the integrity of signaling networks. In this work we have combined biophysical simulations of the Hsp90 crystal structures with the protein structure network analysis to characterize the statistical ensemble of allosteric interaction networks and communication pathways in the Hsp90 chaperones. We have found that principal structurally stable communities could be preserved during dynamic changes in the conformational ensemble. The dominant contribution of the inter-domain rigidity to the interaction networks has emerged as a common factor responsible for the thermodynamic stability of the active chaperone form during the ATPase cycle. Structural stability analysis using force constant profiling of the inter-residue fluctuation distances has identified a network of conserved structurally rigid residues that could serve as global mediating sites of allosteric communication. Mapping of the conformational landscape with the network centrality parameters has demonstrated that stable communities and mediating residues may act concertedly with the shifts in the conformational equilibrium and could describe the majority of functionally significant chaperone residues. The network analysis has revealed a relationship between structural stability, global centrality and functional significance of hotspot residues involved in chaperone regulation. We have found that allosteric interactions in the Hsp90 chaperone may be mediated by modules of structurally stable residues that display high betweenness in the global interaction network. The results of this study have suggested that allosteric interactions in the Hsp90 chaperone may operate via a mechanism that combines rapid and efficient communication by a single optimal pathway of structurally rigid residues and more robust signal transmission using an ensemble of suboptimal multiple communication routes. This

  19. Real time biomedical signal transmission of mixed ECG signal and patient information using visible light communication.

    PubMed

    Tan, Yee Yong; Jung, Sang-Joong; Chung, Wan-Young

    2013-01-01

    The utilization of radio-frequency (RF) communication technology in healthcare application, especially in the transmission of health-related data such as biomedical signal and patient information is often perturbed by electromagnetic interference (EMI). This will not only significantly reduce the accuracy and reliability of the data transmitted, but could also compromise the safety of the patients due to radio frequency (RF) radiation. In this paper, we propose a method which utilizes visible light communication technology as a platform for transmission and to provide real-time monitoring of heart rate and patient information. White LED beam is used as the illuminating source to simultaneously transmit biomedical signal as well as patient record. On-off Keying (OOK) modulation technique is used to modulate all the data onto the visible light beam. Both types of data will be transmitted using a single data packet. At the receiving end, a receiver circuit consisting of a high-speed PIN photodetector and a demodulation circuit is employed to demodulate the data from the visible light beam. The demodulated data is then serially transmitted to a personal computer where the biomedical signal, patient information and heart rate can be monitored in real-time.

  20. Concentric core optical fiber with multiple-mode signal transmission

    DOEpatents

    Muhs, J.D.

    1997-05-06

    A concentric core optical fiber provides for the simultaneous but independent transmission of signals over a single optical fiber. The concentric optical fiber is constructed of a single-mode or multimode inner optical fiber defined by a core and a cladding of a lower index of refraction than the core and an outer optical fiber defined by additional cladding concentrically disposed around the cladding and of an index of refraction lower than the first mentioned cladding whereby the latter functions as the core of the outer optical fiber. By employing such an optical fiber construction with a single-mode inner core or optical fiber, highly sensitive interferometric and stable less sensitive amplitude based sensors can be placed along the same length of a concentric core optical fiber. Also, by employing the concentric core optical fiber secure telecommunications can be achieved via the inner optical fiber since an intrusion of the concentric optical fiber will first cause a variation in the light being transmitted through the outer optical fiber and this variation of light being used to trigger a suitable alarm indicative of the intrusion. 3 figs.

  1. Concentric core optical fiber with multiple-mode signal transmission

    DOEpatents

    Muhs, Jeffrey D.

    1997-01-01

    A concentric core optical fiber provides for the simultaneous but independent transmission of signals over a single optical fiber. The concentric optical fiber is constructed of a single-mode or multimode inner optical fiber defined by a core and a cladding of a lower index of refraction than the core and an outer optical fiber defined by additional cladding concentrically disposed around the cladding and of an index of refraction lower than the first mentioned cladding whereby the latter functions as the core of the outer optical fiber. By employing such an optical fiber construction with a single-mode inner core or optical fiber, highly sensitive interferometric and stable less sensitive amplitude based sensors can be placed along the same length of a concentric core optical fiber. Also, by employing the concentric core optical fiber secure telecommunications can be achieved via the inner optical fiber since an intrusion of the concentric optical fiber will first cause a variation in the light being transmitted through the outer optical fiber and this variation of light being used to trigger a suitable alarm indicative of the intrusion.

  2. INSTRUMENTATION FOR SURVEYING ACOUSTIC SIGNALS IN NATURAL GAS TRANSMISSION LINES

    SciTech Connect

    John L. Loth; Gary J. Morris; George M. Palmer; Richard Guiler; Deepak Mehra

    2003-09-01

    In the U.S. natural gas is distributed through more than one million miles of high-pressure transmission pipelines. If all leaks and infringements could be detected quickly, it would enhance safety and U.S. energy security. Only low frequency acoustic waves appear to be detectable over distances up to 60 km where pipeline shut-off valves provide access to the inside of the pipeline. This paper describes a Portable Acoustic Monitoring Package (PAMP) developed to record and identify acoustic signals characteristic of: leaks, pump noise, valve and flow metering noise, third party infringement, manual pipeline water and gas blow-off, etc. This PAMP consists of a stainless steel 1/2 inch NPT plumbing tree rated for use on 1000 psi pipelines. Its instrumentation is designed to measure acoustic waves over the entire frequency range from zero to 16,000 Hz by means of four instruments: (1) microphone, (2) 3-inch water full range differential pressure transducer with 0.1% of range sensitivity, (3) a novel 3 inch to 100 inch water range amplifier, using an accumulator with needle valve and (4) a line-pressure transducer. The weight of the PAMP complete with all accessories is 36 pounds. This includes a remote control battery/switch box assembly on a 25-foot extension chord, a laptop data acquisition computer on a field table and a sun shield.

  3. Optical signal monitoring in phase modulated optical fiber transmission systems

    NASA Astrophysics Data System (ADS)

    Zhao, Jian

    Optical performance monitoring (OPM) is one of the essential functions for future high speed optical networks. Among the parameters to be monitored, chromatic dispersion (CD) is especially important since it has a significant impact on overall system performance. In this thesis effective CD monitoring approaches for phase-shift keying (PSK) based optical transmission systems are investigated. A number of monitoring schemes based on radio frequency (RF) spectrum analysis and delay-tap sampling are proposed and their performance evaluated. A method for dispersion monitoring of differential phase-shift keying (DPSK) signals based on RF power detection is studied. The RF power spectrum is found to increase with the increase of CD and decrease with polarization mode dispersion (PMD). The spectral power density dependence on CD is studied theoretically and then verified through simulations and experiments. The monitoring sensitivity for nonreturn-to-zero differential phase-shift keying (NRZ-DPSK) and return-to-zero differential phase-shift keying (RZ-DPSK) based systems can reach 80ps/nm/dB and 34ps/nm/dB respectively. The scheme enables the monitoring of differential group delay (DGD) and CD simultaneously. The monitoring sensitivity of CD and DGD can reach 56.7ps/nm/dB and 3.1ps/dB using a bandpass filter. The effects of optical signal-to-noise ratio (OSNR), DGD, fiber nonlinearity and chirp on the monitoring results are investigated. Two RF pilot tones are employed for CD monitoring of DPSK signals. Specially selected pilot tone frequencies enable good monitoring sensitivity with minimum influence on the received signals. The dynamic range exceeding 35dB and monitoring sensitivity up to 9.5ps/nm/dB are achieved. Asynchronous sampling technique is employed for CD monitoring. A signed CD monitoring method for 10Gb/s NRZ-DPSK and RZ-DPSK systems using asynchronous delay-tap sampling technique is studied. The demodulated signals suffer asymmetric waveform distortion if

  4. Allosteric Modulators for the Treatment of Schizophrenia: Targeting Glutamatergic Networks

    PubMed Central

    Menniti, Frank S.; Lindsley, Craig W.; Conn, P. Jeffrey; Pandit, Jayvardhan; Zagouras, Panayiotis; Volkmann, Robert A.

    2013-01-01

    Schizophrenia is a highly debilitating mental disorder which afflicts approximately 1% of the global population. Cognitive and negative deficits account for the lifelong disability associated with schizophrenia, whose symptoms are not effectively addressed by current treatments. New medicines are needed to treat these aspects of the disease. Neurodevelopmental, neuropathological, genetic, and behavioral pharmacological data indicate that schizophrenia stems from a dysfunction of glutamate synaptic transmission, particularly in frontal cortical networks. A number of novel pre- and postsynaptic mechanisms affecting glutamatergic synaptic transmission have emerged as viable targets for schizophrenia. While developing orthosteric glutamatergic agents for these targets has proven extremely difficult, targeting allosteric sites of these targets has emerged as a promising alternative. From a medicinal chemistry perspective, allosteric sites provide an opportunity of finding agents with better drug-like properties and greater target specificity. Furthermore, allosteric modulators are better suited to maintaining the highly precise temporal and spatial aspects of glutamatergic synaptic transmission. Herein, we review neuropathological and genomic/genetic evidence underscoring the importance of glutamate synaptic dysfunction in the etiology of schizophrenia and make a case for allosteric targets for therapeutic intervention. We review progress in identifying allosteric modulators of AMPA receptors, NMDA receptors, and metabotropic glutamate receptors, all with the aim of restoring physiological glutamatergic synaptic transmission. Challenges remain given the complexity of schizophrenia and the difficulty in studying cognition in animals and humans. Nonetheless, important compounds have emerged from these efforts and promising preclinical and variable clinical validation has been achieved. PMID:23409764

  5. Mapping of the Allosteric Site in Cholesterol Hydroxylase CYP46A1 for Efavirenz, a Drug That Stimulates Enzyme Activity.

    PubMed

    Anderson, Kyle W; Mast, Natalia; Hudgens, Jeffrey W; Lin, Joseph B; Turko, Illarion V; Pikuleva, Irina A

    2016-05-27

    Cytochrome P450 46A1 (CYP46A1) is a microsomal enzyme and cholesterol 24-hydroxylase that controls cholesterol elimination from the brain. This P450 is also a potential target for Alzheimer disease because it can be activated pharmacologically by some marketed drugs, as exemplified by efavirenz, the anti-HIV medication. Previously, we suggested that pharmaceuticals activate CYP46A1 allosterically through binding to a site on the cytosolic protein surface, which is different from the enzyme active site facing the membrane. Here we identified this allosteric site for efavirenz on CYP46A1 by using a combination of hydrogen-deuterium exchange coupled to MS, computational modeling, site-directed mutagenesis, and analysis of the CYP46A1 crystal structure. We also mapped the binding region for the CYP46A1 redox partner oxidoreductase and found that the allosteric and redox partner binding sites share a common border. On the basis of the data obtained, we propose the mechanism of CYP46A1 allostery and the pathway for the signal transmission from the P450 allosteric site to the active site.

  6. Development of a high throughput screen for allosteric modulators of melanocortin-4 receptor signaling using a real time cAMP assay

    PubMed Central

    Pantel, Jacques; Williams, Savannah Y.; Mi, Dehui; Sebag, Julien; Corbin, Jackie D.; Weaver, C. David; Cone, Roger D.

    2011-01-01

    The melanocortin MC4 receptor is a potential target for the development of drugs for both obesity and cachexia. Melanocortin MC4 receptor ligands known thus far are orthosteric agonists or antagonists, however the agonists, in particular, have generally exhibited unwanted side effects. For some receptors, allosteric modulators are expected to reduce side-effect profiles. To identify allosteric modulators of the melanocortin MC4 receptor, we created HEK293 cell lines coexpressing the human melanocortin MC4 receptor and a modified luciferase-based cAMP sensor. Monitoring luminescence as a readout of real-time intracellular cAMP concentration, we demonstrate this cell line is able to report melanocortin agonist responses, as well as inverse agonist response to the physiological AgRP peptide. Based on the MC4R-GLO cell line, we developed an assay that was shown to meet HTS standards (Z’=0.50). A pilot screen run on the Microsource Spectrum compound library (n= 2,000) successfully identified 62 positive modulators. This screen identified predicted families of compounds: β2AR agonists –the β2AR being endogenously expressed in HEK293 cells-, an adenylyl cyclase activator and finally a distribution of phosphodiesterase (PDE) inhibitors well characterized or recently identified. In this last category, we identified a structural family of coumarin-derived compounds (imperatorin, osthol and prenyletin), along with deracoxib, a drug in veterinary use for its COX2 inhibitory properties. This latter finding unveiled a new off-target mechanism of action for deracoxib as a PDE inhibitor. Overall, these data are the first report of an HTS for allosteric modulators for a Gs protein coupled receptor. PMID:21296065

  7. A linear signal transmission system calibration method of wideband GPR

    NASA Astrophysics Data System (ADS)

    Wu, Bin; Zhao, Kai; Gu, Ling-jia; Cao, Qiong; Li, Xiao-feng; Zheng, Xing-ming

    2016-09-01

    In VHF pulse Ground Penetrating Radar(GPR) system, the echo pass through the antenna and transmission line circuit, then reach the GPR receiver. Thus the reflection coefficient at the receiver sampling gate interface, which is at the end of the transmission line, is different from the real reflection coefficient of the media at the antenna interface, which could cause the GPR receiving error. The pulse GPR receiver is a wideband system that can't be simply described as traditional narrowband transmission line model. Since the GPR transmission circuit is a linear system, the linear transformation method could be used to analyze the characteristic of the GPR receiving system. A GPR receiver calibration method based on transmission line theory is proposed in this paper, which analyzes the relationship between the reflection coefficients of theory calculation at antenna interface and the measuring data by network analyzer at the sampling gate interface. Then the least square method is introduced to calibrate the transfer function of the GPR receiver transmission circuit. This calibration method can be useful in media quantitative inversion by GPR. When the reflection coefficient at the sampling gate is obtained, the real reflection coefficient of the media at the antenna interface can be easily determined.

  8. Controlling allosteric networks in proteins

    NASA Astrophysics Data System (ADS)

    Dokholyan, Nikolay

    2013-03-01

    We present a novel methodology based on graph theory and discrete molecular dynamics simulations for delineating allosteric pathways in proteins. We use this methodology to uncover the structural mechanisms responsible for coupling of distal sites on proteins and utilize it for allosteric modulation of proteins. We will present examples where inference of allosteric networks and its rewiring allows us to ``rescue'' cystic fibrosis transmembrane conductance regulator (CFTR), a protein associated with fatal genetic disease cystic fibrosis. We also use our methodology to control protein function allosterically. We design a novel protein domain that can be inserted into identified allosteric site of target protein. Using a drug that binds to our domain, we alter the function of the target protein. We successfully tested this methodology in vitro, in living cells and in zebrafish. We further demonstrate transferability of our allosteric modulation methodology to other systems and extend it to become ligh-activatable.

  9. A study on Data Transmission Scheme for High Functional Railway Signaling System

    NASA Astrophysics Data System (ADS)

    Ishikawa, Ryo; Sano, Minoru; Mochizuki, Hiroshi; Nakamura, Hideo

    Railway signaling systems that transmit control information via rails have been deployed in many applications, for example, digital automatic train control (ATC) systems for controlling train speed. Since the performance of digital ATC systems depends on the signal transmission speed, recently there have been many studies aimed at realizing high-speed data transmission. However, it is difficult to increase the transmission speed because rails have strong attenuation in proportion to an increase of the frequency. In this paper, we aimed to increase the transmission speed by improving the modulation scheme to overcome these limitations. We proposed CDMA-QAM method that is combined code-division multiple access (CDMA) and quadrature amplitude modulation (QAM). And we evaluated in a field trial the CDMA-QAM rail transmission device developed using DSP. On the other hand, an analog ATC based on an amplitude modulation (AM) is still employed in some railway lines. It is difficult for their lines to introduce the digital signal due to track circuit configurations and interoperability conditions. So we studied a data transmission scheme that makes it possible to mix an analog signal and a digital signal, and evaluated the influence given to both signals using a developed device that generates the mixed signal of analog and digital signal.

  10. [Research progress on key technology of power and signal transmission in neuroprosthetic].

    PubMed

    Wang, Xing; Peng, Chenglin; Liu, Tao; Wang, Rui; Hou, Wensheng; Zheng, Xiaolin; Zheng, Erxin

    2011-10-01

    The power and signal transmission technology is one of the key technologies in neuroprosthetic research. This paper proposes firstly the related theory of power and signal transmission technology in neuroprosthetic, then summarizes the three key aspects of the power and signal transmission technology in neuroprosthetic. After analyzed the development of the inductive wireless power harvesting technology, the wireless telemetry technology and the wireless power harvesting telemetry technology, the emphasis on research contents will be proposed and discussed, which will help accelerate the further research of prosthetic.

  11. Computational Tools for Allosteric Drug Discovery: Site Identification and Focus Library Design.

    PubMed

    Huang, Wenkang; Nussinov, Ruth; Zhang, Jian

    2017-01-01

    Allostery is an intrinsic phenomenon of biological macromolecules involving regulation and/or signal transduction induced by a ligand binding to an allosteric site distinct from a molecule's active site. Allosteric drugs are currently receiving increased attention in drug discovery because drugs that target allosteric sites can provide important advantages over the corresponding orthosteric drugs including specific subtype selectivity within receptor families. Consequently, targeting allosteric sites, instead of orthosteric sites, can reduce drug-related side effects and toxicity. On the down side, allosteric drug discovery can be more challenging than traditional orthosteric drug discovery due to difficulties associated with determining the locations of allosteric sites and designing drugs based on these sites and the need for the allosteric effects to propagate through the structure, reach the ligand binding site and elicit a conformational change. In this study, we present computational tools ranging from the identification of potential allosteric sites to the design of "allosteric-like" modulator libraries. These tools may be particularly useful for allosteric drug discovery.

  12. Coherent Terahertz Wireless Signal Transmission Using Advanced Optical Fiber Communication Technology

    NASA Astrophysics Data System (ADS)

    Kanno, Atsushi; Kuri, Toshiaki; Morohashi, Isao; Hosako, Iwao; Kawanishi, Tetsuya; Yoshida, Yuki; Kitayama, Ken-ichi

    2015-02-01

    Coherent terahertz signal transmission with multilevel modulation and demodulation is demonstrated using an optical sub-harmonic IQ mixer (SHIQM), which consists of optical components in advanced optical fiber communication technologies. An optical-frequency-comb-employed signal generator is capable of vector modulation as well as frequency tunability. Digital signal processing (DSP) adopted from the recently developed optical digital coherent communication can easily demodulate multi-level modulated terahertz signals by using electrical heterodyning for intermediate-frequency (IF) down conversion. This technique is applicable for mobile backhauling in the next-generation mobile communication technology directly connected to an optical fiber network as a high-speed wireless transmission link.

  13. Method and apparatus for low-loss signal transmission

    NASA Technical Reports Server (NTRS)

    Siegel, Peter (Inventor); Yeh, Cavour (Inventor); Shimabukuro, Fred (Inventor); Fraser, Scott (Inventor)

    2008-01-01

    The present invention relates to the field of radio-frequency (RF) waveguides. More specifically, the present invention pertains to a method and apparatus that provides ultra-low-loss RF waveguide structures targeted between approximately 300 GHz and approximately 30 THz. The RF waveguide includes a hollow core and a flexible honeycomb, periodic-bandgap structure surrounding the hollow core. The flexible honeycomb, periodic-bandgap structure is formed of a plurality of tubes formed of a dielectric material such as of low-loss quartz, polyethylene, or high-resistivity silicon. Using the RF waveguide, a user may attach a terahertz signal source to the waveguide and pass signals through the waveguide, while a terahertz signal receiver receives the signals.

  14. Signal Detection Theory Applied to Helicopter Transmission Diagnostic Thresholds

    NASA Technical Reports Server (NTRS)

    Dempsey, Paula J.; Keller, Jonathan A.; Wade, Daniel R.

    2008-01-01

    Helicopter Health Usage Monitoring Systems (HUMS) have potential for providing data to support increasing the service life of a dynamic mechanical component in the transmission of a helicopter. Data collected can demonstrate the HUMS condition indicator responds to a specific component fault with appropriate alert limits and minimal false alarms. Defining thresholds for specific faults requires a tradeoff between the sensitivity of the condition indicator (CI) limit to indicate damage and the number of false alarms. A method using Receiver Operating Characteristic (ROC) curves to assess CI performance was demonstrated using CI data collected from accelerometers installed on several UH60 Black Hawk and AH64 Apache helicopters and an AH64 helicopter component test stand. Results of the analysis indicate ROC curves can be used to reliably assess the performance of commercial HUMS condition indicators to detect damaged gears and bearings in a helicopter transmission.

  15. Signal Detection Theory Applied to Helicopter Transmission Diagnostic Thresholds

    NASA Technical Reports Server (NTRS)

    Dempsey, Paula J.; Keller, Jonathan A.; Wade, Daniel R.

    2009-01-01

    Helicopter Health Usage Monitoring Systems (HUMS) have potential for providing data to support increasing the service life of a dynamic mechanical component in the transmission of a helicopter. Data collected can demonstrate the HUMS condition indicator responds to a specific component fault with appropriate alert limits and minimal false alarms. Defining thresholds for specific faults requires a tradeoff between the sensitivity of the condition indicator (CI) limit to indicate damage and the number of false alarms. A method using Receiver Operating Characteristic (ROC) curves to assess CI performance was demonstrated using CI data collected from accelerometers installed on several UH60 Black Hawk and AH64 Apache helicopters and an AH64 helicopter component test stand. Results of the analysis indicate ROC curves can be used to reliably assess the performance of commercial HUMS condition indicators to detect damaged gears and bearings in a helicopter transmission.

  16. Paired SSB optical OFDM channels for high spectral efficient signal transmission over DWDM networks

    NASA Astrophysics Data System (ADS)

    Chicharro, Francisco I.; Ortega, Beatriz; Mora, José

    2016-07-01

    A new high spectral efficient SSB-OOFDM DWDM transmission system has been experimentally demonstrated. The proposed transmitter employs paired optical channels consisting of two SSB modulated OFDM signals using opposite sidebands in order to allow an efficient use of the spectrum with optical carriers separation under 10 GHz. Moreover, different paired channels are multiplexed into the 25 GHz grid DWDM fiber transmission link. Optical carrier spacing of 8.75 GHz in paired channels has been demonstrated allowing 40.8 Gb/s signal transmission rate over a 25 GHz paired channel bandwidth.

  17. Efficient Sparse Signal Transmission over a Lossy Link Using Compressive Sensing

    PubMed Central

    Wu, Liantao; Yu, Kai; Cao, Dongyu; Hu, Yuhen; Wang, Zhi

    2015-01-01

    Reliable data transmission over lossy communication link is expensive due to overheads for error protection. For signals that have inherent sparse structures, compressive sensing (CS) is applied to facilitate efficient sparse signal transmissions over lossy communication links without data compression or error protection. The natural packet loss in the lossy link is modeled as a random sampling process of the transmitted data, and the original signal will be reconstructed from the lossy transmission results using the CS-based reconstruction method at the receiving end. The impacts of packet lengths on transmission efficiency under different channel conditions have been discussed, and interleaving is incorporated to mitigate the impact of burst data loss. Extensive simulations and experiments have been conducted and compared to the traditional automatic repeat request (ARQ) interpolation technique, and very favorable results have been observed in terms of both accuracy of the reconstructed signals and the transmission energy consumption. Furthermore, the packet length effect provides useful insights for using compressed sensing for efficient sparse signal transmission via lossy links. PMID:26287195

  18. Electronic post-compensation of WDM transmission impairments using coherent detection and digital signal processing.

    PubMed

    Li, Xiaoxu; Chen, Xin; Goldfarb, Gilad; Mateo, Eduardo; Kim, Inwoong; Yaman, Fatih; Li, Guifang

    2008-01-21

    A universal post-compensation scheme for fiber impairments in wavelength-division multiplexing (WDM) systems is proposed based on coherent detection and digital signal processing (DSP). Transmission of 10 x 10 Gbit/s binary-phase-shift-keying (BPSK) signals at a channel spacing of 20 GHz over 800 km dispersion shifted fiber (DSF) has been demonstrated numerically.

  19. Navigation Using Signals of Opportunity in the AM Transmission Band

    DTIC Science & Technology

    2006-09-01

    compo- nents. The overall goal was to create two signals, both sampled at the standard GNURadio sampling frequency of 4MHz, which are separated in...detailed in the previous section, the USRP hardware, the GNURadio software detailed in Section 2.5, two personal computers, and various cables and...the following World Wide Web (WWW) sites are highly recommended: 1. http://www.gnu.org/software/ gnuradio /index.html 2. http://www.nd.edu/~dshen/GNU/ 3

  20. Secure chaotic transmission of electrocardiography signals with acousto-optic modulation under profiled beam propagation.

    PubMed

    Almehmadi, Fares S; Chatterjee, Monish R

    2015-01-10

    Electrocardiography (ECG) signals are used for both medical purposes and identifying individuals. It is often necessary to encrypt this highly sensitive information before it is transmitted over any channel. A closed-loop acousto-optic hybrid device acting as a chaotic modulator is applied to ECG signals to achieve this encryption. Recently improved modeling of this approach using profiled optical beams has shown it to be very sensitive to key parameters that characterize the encryption and decryption process, exhibiting its potential for secure transmission of analog and digital signals. Here the encryption and decryption is demonstrated for ECG signals, both analog and digital versions, illustrating strong encryption without significant distortion. Performance analysis pertinent to both analog and digital transmission of the ECG waveform is also carried out using output signal-to-noise, signal-to-distortion, and bit-error-rate measures relative to the key parameters and presence of channel noise in the system.

  1. High-quality frame-synchronization for satellite video signal transmission

    NASA Astrophysics Data System (ADS)

    Kubota, Shuji; Morikura, Masahiro; Kato, Shuzo

    1995-01-01

    A high-quality frame-synchronizer for video signal switching and freezing is proposed. In order to realize high-quality frame-synchronization, a novel high-speed and high-definition 11 bit analog-to-digital (A/D) converter which achieves the quite high unweighted S/N ratio performance of 63 dB is developed. It provides synchronized video signal switching by field freezing for high-quality video signal transmission.

  2. Differentiating a Ligand's Chemical Requirements for Allosteric Interactions from Those for Protein Binding. Phenylalanine Inhibition of Pyruvate Kinase

    SciTech Connect

    Williams,R.; Holyoak, T.; McDonald, G.; Gui, C.; Fenton, A.

    2006-01-01

    The isoform of pyruvate kinase from brain and muscle of mammals (M1-PYK) is allosterically inhibited by phenylalanine. Initial observations in this model allosteric system indicate that Ala binds competitively with Phe, but elicits a minimal allosteric response. Thus, the allosteric ligand of this system must have requirements for eliciting an allosteric response in addition to the requirements for binding. Phe analogues have been used to dissect what chemical properties of Phe are responsible for eliciting the allosteric response. We first demonstrate that the L-2-aminopropanaldehyde substructure of the amino acid ligand is primarily responsible for binding to M1-PYK. Since the allosteric response to Ala is minimal and linear addition of methyl groups beyond the -carbon increase the magnitude of the allosteric response, we conclude that moieties beyond the -carbon are primarily responsible for allostery. Instead of an all-or-none mechanism of allostery, these findings support the idea that the bulk of the hydrophobic side chain, but not the aromatic nature, is the primary determinant of the magnitude of the observed allosteric inhibition. The use of these results to direct structural studies has resulted in a 1.65 Angstroms structure of M1-PYK with Ala bound. The coordination of Ala in the allosteric amino acid binding site confirms the binding role of the L-2-aminopropanaldehyde substructure of the ligand. Collectively, this study confirms that a ligand can have chemical regions specific for eliciting the allosteric signal in addition to the chemical regions necessary for binding.

  3. Leaky coaxial cable signal transmission for remote facilities

    SciTech Connect

    Smith, S.F.; Crutcher, R.I.

    1993-03-01

    To develop reliable communications methods to meet the rigorous requirements for nuclear hot cells and similar environments, including control of cranes, transporters, and advanced servomanipulators, the Consolidated Fuel Reprocessing Program (CFRP) at Oak Ridge National Laboratory (ORNL) has conducted extensive tests of numerous technologies to determine their applicability to remote operations. To alleviate the need for large bundles of cables that must accommodate crane/transporter motion relative to the boundaries of the cell, several transmission techniques are available, including slotted-line radio-frequency couplers, infrared beams, fiber-optic cables, free-space microwave, and inductively coupled leaky coaxial cable. This paper discusses the general characteristics, mode of operation, and proposed implementation of leaky coaxial cable technology in a waste-handling facility scheduled to be built in the near future at ORNL. In addition, specific system hardware based around the use of leaky coaxial cable is described in detail. Finally, data from a series of radiation exposure tests conducted by the CFRP on several samples of the basic leaky coaxial cable and associated connectors are presented.

  4. Leaky coaxial cable signal transmission for remote facilities

    SciTech Connect

    Smith, S.F.; Crutcher, R.I.

    1993-01-01

    To develop reliable communications methods to meet the rigorous requirements for nuclear hot cells and similar environments, including control of cranes, transporters, and advanced servomanipulators, the Consolidated Fuel Reprocessing Program (CFRP) at Oak Ridge National Laboratory (ORNL) has conducted extensive tests of numerous technologies to determine their applicability to remote operations. To alleviate the need for large bundles of cables that must accommodate crane/transporter motion relative to the boundaries of the cell, several transmission techniques are available, including slotted-line radio-frequency couplers, infrared beams, fiber-optic cables, free-space microwave, and inductively coupled leaky coaxial cable. This paper discusses the general characteristics, mode of operation, and proposed implementation of leaky coaxial cable technology in a waste-handling facility scheduled to be built in the near future at ORNL. In addition, specific system hardware based around the use of leaky coaxial cable is described in detail. Finally, data from a series of radiation exposure tests conducted by the CFRP on several samples of the basic leaky coaxial cable and associated connectors are presented.

  5. 37 CFR 258.4 - Royalty fee for secondary transmission of digital signals of broadcast stations by satellite...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... transmission of digital signals of broadcast stations by satellite carriers. 258.4 Section 258.4 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.4 Royalty fee for secondary transmission of digital signals of broadcast stations by satellite carriers....

  6. 37 CFR 258.3 - Royalty fee for secondary transmission of analog signals of broadcast stations by satellite...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... transmission of analog signals of broadcast stations by satellite carriers. 258.3 Section 258.3 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.3 Royalty fee for secondary transmission of analog signals of broadcast stations by satellite carriers....

  7. 37 CFR 258.3 - Royalty fee for secondary transmission of analog signals of broadcast stations by satellite...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... transmission of analog signals of broadcast stations by satellite carriers. 258.3 Section 258.3 Patents... RULES AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.3 Royalty fee for secondary transmission of analog signals of broadcast stations by satellite carriers....

  8. 37 CFR 258.4 - Royalty fee for secondary transmission of digital signals of broadcast stations by satellite...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... transmission of digital signals of broadcast stations by satellite carriers. 258.4 Section 258.4 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.4 Royalty fee for secondary transmission of digital signals of broadcast stations by satellite carriers....

  9. 37 CFR 258.3 - Royalty fee for secondary transmission of analog signals of broadcast stations by satellite...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... transmission of analog signals of broadcast stations by satellite carriers. 258.3 Section 258.3 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.3 Royalty fee for secondary transmission of analog signals of broadcast stations by satellite carriers....

  10. 37 CFR 258.3 - Royalty fee for secondary transmission of analog signals of broadcast stations by satellite...

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... transmission of analog signals of broadcast stations by satellite carriers. 258.3 Section 258.3 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.3 Royalty fee for secondary transmission of analog signals of broadcast stations by satellite carriers....

  11. 37 CFR 258.4 - Royalty fee for secondary transmission of digital signals of broadcast stations by satellite...

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... transmission of digital signals of broadcast stations by satellite carriers. 258.4 Section 258.4 Patents... RULES AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.4 Royalty fee for secondary transmission of digital signals of broadcast stations by satellite carriers....

  12. 37 CFR 258.3 - Royalty fee for secondary transmission of analog signals of broadcast stations by satellite...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... transmission of analog signals of broadcast stations by satellite carriers. 258.3 Section 258.3 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.3 Royalty fee for secondary transmission of analog signals of broadcast stations by satellite carriers....

  13. 37 CFR 258.4 - Royalty fee for secondary transmission of digital signals of broadcast stations by satellite...

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... transmission of digital signals of broadcast stations by satellite carriers. 258.4 Section 258.4 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.4 Royalty fee for secondary transmission of digital signals of broadcast stations by satellite carriers....

  14. 37 CFR 258.4 - Royalty fee for secondary transmission of digital signals of broadcast stations by satellite...

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... transmission of digital signals of broadcast stations by satellite carriers. 258.4 Section 258.4 Patents... AND PROCEDURES ADJUSTMENT OF ROYALTY FEE FOR SECONDARY TRANSMISSIONS BY SATELLITE CARRIERS § 258.4 Royalty fee for secondary transmission of digital signals of broadcast stations by satellite carriers....

  15. Immune response and insulin signalling alter mosquito feeding behaviour to enhance malaria transmission potential

    PubMed Central

    Cator, Lauren J.; Pietri, Jose E.; Murdock, Courtney C.; Ohm, Johanna R.; Lewis, Edwin E.; Read, Andrew F.; Luckhart, Shirley; Thomas, Matthew B.

    2015-01-01

    Malaria parasites alter mosquito feeding behaviour in a way that enhances parasite transmission. This is widely considered a prime example of manipulation of host behaviour to increase onward transmission, but transient immune challenge in the absence of parasites can induce the same behavioural phenotype. Here, we show that alterations in feeding behaviour depend on the timing and dose of immune challenge relative to blood ingestion and that these changes are functionally linked to changes in insulin signalling in the mosquito gut. These results suggest that altered phenotypes derive from insulin signalling-dependent host resource allocation among immunity, blood feeding, and reproduction in a manner that is not specific to malaria parasite infection. We measured large increases in mosquito survival and subsequent transmission potential when feeding patterns are altered. Leveraging these changes in physiology, behaviour and life history could promote effective and sustainable control of female mosquitoes responsible for transmission. PMID:26153094

  16. Immune response and insulin signalling alter mosquito feeding behaviour to enhance malaria transmission potential.

    PubMed

    Cator, Lauren J; Pietri, Jose E; Murdock, Courtney C; Ohm, Johanna R; Lewis, Edwin E; Read, Andrew F; Luckhart, Shirley; Thomas, Matthew B

    2015-07-08

    Malaria parasites alter mosquito feeding behaviour in a way that enhances parasite transmission. This is widely considered a prime example of manipulation of host behaviour to increase onward transmission, but transient immune challenge in the absence of parasites can induce the same behavioural phenotype. Here, we show that alterations in feeding behaviour depend on the timing and dose of immune challenge relative to blood ingestion and that these changes are functionally linked to changes in insulin signalling in the mosquito gut. These results suggest that altered phenotypes derive from insulin signalling-dependent host resource allocation among immunity, blood feeding, and reproduction in a manner that is not specific to malaria parasite infection. We measured large increases in mosquito survival and subsequent transmission potential when feeding patterns are altered. Leveraging these changes in physiology, behaviour and life history could promote effective and sustainable control of female mosquitoes responsible for transmission.

  17. An allosteric role for receptor activity-modifying proteins in defining GPCR pharmacology

    PubMed Central

    J Gingell, Joseph; Simms, John; Barwell, James; Poyner, David R; Watkins, Harriet A; Pioszak, Augen A; Sexton, Patrick M; Hay, Debbie L

    2016-01-01

    G protein-coupled receptors are allosteric proteins that control transmission of external signals to regulate cellular response. Although agonist binding promotes canonical G protein signalling transmitted through conformational changes, G protein-coupled receptors also interact with other proteins. These include other G protein-coupled receptors, other receptors and channels, regulatory proteins and receptor-modifying proteins, notably receptor activity-modifying proteins (RAMPs). RAMPs have at least 11 G protein-coupled receptor partners, including many class B G protein-coupled receptors. Prototypic is the calcitonin receptor, with altered ligand specificity when co-expressed with RAMPs. To gain molecular insight into the consequences of this protein–protein interaction, we combined molecular modelling with mutagenesis of the calcitonin receptor extracellular domain, assessed in ligand binding and functional assays. Although some calcitonin receptor residues are universally important for peptide interactions (calcitonin, amylin and calcitonin gene-related peptide) in calcitonin receptor alone or with receptor activity-modifying protein, others have RAMP-dependent effects, whereby mutations decreased amylin/calcitonin gene-related peptide potency substantially only when RAMP was present. Remarkably, the key residues were completely conserved between calcitonin receptor and AMY receptors, and between subtypes of AMY receptor that have different ligand preferences. Mutations at the interface between calcitonin receptor and RAMP affected ligand pharmacology in a RAMP-dependent manner, suggesting that RAMP may allosterically influence the calcitonin receptor conformation. Supporting this, molecular dynamics simulations suggested that the calcitonin receptor extracellular N-terminal domain is more flexible in the presence of receptor activity-modifying protein 1. Thus, RAMPs may act in an allosteric manner to generate a spectrum of unique calcitonin receptor

  18. LVDS tester: a systematic test of cable signal transmission at the ALICE experiment

    NASA Astrophysics Data System (ADS)

    Barnby, L.; Bhasin, A.; Bombara, M.; Evans, D.; Jones, G. T.; Jones, P. G.; Jovanović, P.; Jusko, A.; Kour, R.; Králik, I.; Krivda, M.; Lazzeroni, C.; Lietava, R.; Matthews, Z. L.; Navin, S.; Palaha, A.; Petrov, P.; Platt, R.; Šándor, L.; Scott, P.; Urbán, J.; Villalobos Baillie, O.; Tapia Takaki, J. D.

    2010-12-01

    In the ALICE experiment, the Low-Voltage Differential Signalling (LVDS) format is used for the transmission of trigger inputs from the detectors to the Central Trigger Processor (CTP), the L0 trigger outputs from Local Trigger Units (LTU) boards back to the detectors and the BUSY inputs from the sub-detectors to the CTP. ALICE has designed a set-up, called the LVDS transmission tester, that aims to measure various transmission quality parameters and the bit-error rate (BER) for long period runs in an automatic way. In this paper, this method is described and the conclusions from these tests for the ALICE LVDS cables are discussed.

  19. Demonstration of optical steganography transmission using temporal phase coded optical signals with spectral notch filtering.

    PubMed

    Hong, Xuezhi; Wang, Dawei; Xu, Lei; He, Sailing

    2010-06-07

    A novel approach is proposed and experimentally demonstrated for optical steganography transmission in WDM networks using temporal phase coded optical signals with spectral notch filtering. A temporal phase coded stealth channel is temporally and spectrally overlaid onto a public WDM channel. Direct detection of the public channel is achieved in the presence of the stealth channel. The interference from the public channel is suppressed by spectral notching before the detection of the optical stealth signal. The approach is shown to have good compatibility and robustness to the existing WDM network for optical steganography transmission.

  20. Extremely wideband signal shaping using one- and two-dimensional nonuniform nonlinear transmission lines

    NASA Astrophysics Data System (ADS)

    Afshari, E.; Bhat, H. S.; Hajimiri, A.; Marsden, J. E.

    2006-03-01

    We propose a class of electrical circuits for extremely wideband (EWB) signal shaping. A one-dimensional, nonlinear, nonuniform transmission line is proposed for narrow pulse generation. A two-dimensional transmission lattice is proposed for EWB signal combining. Model equations for the circuits are derived. Theoretical and numerical solutions of the model equations are presented, showing that the circuits can be used for the desired application. The procedure by which the circuits are designed exemplifies a modern, mathematical design methodology for EWB circuits.

  1. Cholesterol-mediated allosteric regulation of the mitochondrial translocator protein structure

    PubMed Central

    Jaipuria, Garima; Leonov, Andrei; Giller, Karin; Vasa, Suresh Kumar; Jaremko, Łukasz; Jaremko, Mariusz; Linser, Rasmus; Becker, Stefan; Zweckstetter, Markus

    2017-01-01

    Cholesterol is an important regulator of membrane protein function. However, the exact mechanisms involved in this process are still not fully understood. Here we study how the tertiary and quaternary structure of the mitochondrial translocator protein TSPO, which binds cholesterol with nanomolar affinity, is affected by this sterol. Residue-specific analysis of TSPO by solid-state NMR spectroscopy reveals a dynamic monomer–dimer equilibrium of TSPO in the membrane. Binding of cholesterol to TSPO's cholesterol-recognition motif leads to structural changes across the protein that shifts the dynamic equilibrium towards the translocator monomer. Consistent with an allosteric mechanism, a mutation within the oligomerization interface perturbs transmembrane regions located up to 35 Å away from the interface, reaching TSPO's cholesterol-binding motif. The lower structural stability of the intervening transmembrane regions provides a mechanistic basis for signal transmission. Our study thus reveals an allosteric signal pathway that connects membrane protein tertiary and quaternary structure with cholesterol binding. PMID:28358007

  2. Physical limits on computation by assemblies of allosteric proteins

    NASA Astrophysics Data System (ADS)

    Robinson, John

    2009-03-01

    Assemblies of allosteric proteins are the principle information processing devices in biology. Using the Ca^2+-sensitive cardiac regulatory assembly as a paradigm for Brownian computation, we examine how system complexity and system resetting impose physical limits on computation. Nearest-neighbor-limited interactions among assembly components constrains the topology of the system's macrostate free energy landscape and produces degenerate transition probabilities. As a result, signaling fidelity and deactivation kinetics can not be simultaneously optimized. This imposes an upper limit on the rate of information processing by assemblies of allosteric proteins that couple to a single ligand type.

  3. Physical Limits on Computation by Assemblies of Allosteric Proteins

    NASA Astrophysics Data System (ADS)

    Robinson, John M.

    2008-10-01

    Assemblies of allosteric proteins are the principle information processing devices in biology. Using the Ca2+-sensitive cardiac regulatory assembly as a paradigm for Brownian computation, I examine how system complexity and system resetting impose physical limits on computation. Nearest-neighbor-limited interactions among assembly components constrain the topology of the system’s macrostate free energy landscape and produce degenerate transition probabilities. As a result, signaling fidelity and deactivation kinetics cannot be simultaneously optimized. This imposes an upper limit on the rate of information processing by assemblies of allosteric proteins that couple to a single ligand type.

  4. [Study on signal transmission characteristics of meridian based on electrical network theory and experiments].

    PubMed

    Wang, Zhi-Gong; Lü, Xiao-Ying; Gao, Jian-Yun; Wang, Yu-Hang; Huang, Cen-Yu; Chen, Yue-Lin; Xing, Li-Yang; Wang, Gui-Ying

    2011-08-01

    Study on features of acupoints with resistance test in the past half century is reviewed in this article. Mechanism and technology of the method are introduced as well as its shortcomings. The determination method of signal transmission along meridians with the combination of electrical network theories and practice is advanced. And the result of a series experiments on one meridian at the superficial part of the body are given as well. Thus, it is concluded that the signals of the point-in/point-out and the signals along a non-meridian path with the same distance are significantly different, which gives a verification of the feasibility of the method by using electrical network theories to set out characteristics of signal transmission along meridians dynamically.

  5. Transmission line galloping image monitoring system based on digital signal processor

    NASA Astrophysics Data System (ADS)

    Ren, Hai Peng; Ma, Zhan Feng

    2011-06-01

    An embedded image monitoring system based on TMS320DM642 Digital Signal Processor (DSP) is proposed for the transmission line monitoring. The system can detect galloping, ice or snow covering, and other abnormal status of the transmission line in a real time mode. The image detection algorithms are compared using the controlled experiment under the complex weather environment, thereby, a set of image processing algorithms is proposed for transmission lines image monitoring. The DSP/BOIS multi-threaded programming techniques are used to realize the algorithm in the DSPs' embedded software. A wireless communication based on General Packet Radio Service (GPRS) module is designed to transmit the detection results and the changed information of the image to the monitoring center, so that the operators can get the real time status of the transmission line. The application of the system will play an important role in the condition-based maintenance of power transmission lines and improve the reliability of power delivery system. Transmission line; Status monitoring; Complex weather factor; Image filtering and sharpening; Image segment; Morphological filtering; Wireless communication; Digital signal processor; Multi-threaded programming.

  6. Performance Analysis of Control Signal Transmission Technique for Cognitive Radios in Dynamic Spectrum Access Networks

    NASA Astrophysics Data System (ADS)

    Sakata, Ren; Tomioka, Tazuko; Kobayashi, Takahiro

    When cognitive radio (CR) systems dynamically use the frequency band, a control signal is necessary to indicate which carrier frequencies are currently available in the network. In order to keep efficient spectrum utilization, this control signal also should be transmitted based on the channel conditions. If transmitters dynamically select carrier frequencies, receivers have to receive control signals without knowledge of their carrier frequencies. To enable such transmission and reception, this paper proposes a novel scheme called DCPT (Differential Code Parallel Transmission). With DCPT, receivers can receive low-rate information with no knowledge of the carrier frequencies. The transmitter transmits two signals whose carrier frequencies are spaced by a predefined value. The absolute values of the carrier frequencies can be varied. When the receiver acquires the DCPT signal, it multiplies the signal by a frequency-shifted version of the signal; this yields a DC component that represents the data signal which is then demodulated. The performance was evaluated by means of numerical analysis and computer simulation. We confirmed that DCPT operates successfully even under severe interference if its parameters are appropriately configured.

  7. Experimental demonstration of analog signal transmission in a silicon photonic crystal L3 resonator.

    PubMed

    Gui, Chengcheng; Zhang, Yong; Du, Jing; Xia, Jinsong; Wang, Jian

    2015-06-01

    We design and fabricate a silicon photonic crystal L3 resonator for chip-scale analog signal transmission. The lattice constant (a) is 420 nm, and the radius of holes (r) is 126 nm. The three holes adjacent to the cavity are laterally shifted by 0.175a, 0.025a and 0.175a, respectively. We experimentally evaluate the performance of silicon photonic crystal L3 resonator for chip-scale analog signal transmission. The spurious free dynamic ranges (SFDRs) of the second-order harmonic distortion (SHD) and the third-order harmonic distortion (THD), which are important factors to assess the analog link performance, are measured for the chip-scale analog signal transmission through the fabricated silicon photonic crystal L3 resonator. The SHD SFDR and THD SFDR are measured to be ~34.6 dB and ~52.2 dB even with the input optical carrier sitting at the dip resonance wavelength of the fabricated silicon photonic crystal L3 resonator. The influences of the optical carrier wavelength and input optical power on the SHD SFDR and THD SFDR are studied in the experiment. The impacts of geometric parameters of the cavity structure (lattice constant, radius of holes, shift of the hole) on the analog signal transmission are also analyzed, showing favorable analog link performance with relatively large fabrication tolerance to design parameters.

  8. Specific features of waveguide heating due to transmission of high-power microwave signals

    NASA Astrophysics Data System (ADS)

    Kudryavtsev, I. V.; Gotselyuk, O. B.; Novikov, E. S.; Demin, V. G.

    2017-01-01

    Waveguide heating due to transmission of microwave signals is studied. Mathematical models are developed to evaluate heat liberation, and differential equations of thermal balance are derived with allowance for different working conditions of waveguides. The results prove the necessity of the further study of the effect of heat liberation in waveguides on strength and functional characteristics.

  9. Signal transmission through human muscle for implantable medical devices using galvanic intra-body communication technique.

    PubMed

    Chen, Xi Mei; Mak, Peng Un; Pun, Sio Hang; Gao, Yue Ming; Vai, Mang I; Du, Min

    2012-01-01

    Signal transmission over human tissues has long been the center research topic for biomedical engineering in both academic and industrial arenas. This is particular important for implantable medical devices (IMD) to communicate with other sensor devices in achieving health care and monitoring functions. Traditional Radio Frequency (RF) transmission technique suffers from not only high attenuation but also potential interference & eavesdropping. This paper has examined the alternate galvanic type Intra-Body Communication Technique (IBC) in transmitting signal across the body tissue (mainly muscle) in both analytical electromagnetic model with simulation results. Comparisons of these results with traditional RF data in literatures show a high promising potential (saving over 10 dB or more in path loss) for IBC transmission. Concrete discussions and several further research directions are also given out at the end of this paper.

  10. The Second Extracellular Loop of the Adenosine A1 Receptor Mediates Activity of Allosteric Enhancers

    PubMed Central

    Kennedy, Dylan P.; McRobb, Fiona M.; Leonhardt, Susan A.; Purdy, Michael; Figler, Heidi; Marshall, Melissa A.; Chordia, Mahendra; Figler, Robert; Linden, Joel

    2014-01-01

    Allosteric enhancers of the adenosine A1 receptor amplify signaling by orthosteric agonists. Allosteric enhancers are appealing drug candidates because their activity requires that the orthosteric site be occupied by an agonist, thereby conferring specificity to stressed or injured tissues that produce adenosine. To explore the mechanism of allosteric enhancer activity, we examined their action on several A1 receptor constructs, including (1) species variants, (2) species chimeras, (3) alanine scanning mutants, and (4) site-specific mutants. These findings were combined with homology modeling of the A1 receptor and in silico screening of an allosteric enhancer library. The binding modes of known docked allosteric enhancers correlated with the known structure-activity relationship, suggesting that these allosteric enhancers bind to a pocket formed by the second extracellular loop, flanked by residues S150 and M162. We propose a model in which this vestibule controls the entry and efflux of agonists from the orthosteric site and agonist binding elicits a conformational change that enables allosteric enhancer binding. This model provides a mechanism for the observations that allosteric enhancers slow the dissociation of orthosteric agonists but not antagonists. PMID:24217444

  11. Power-efficient method for IM-DD optical transmission of multiple OFDM signals.

    PubMed

    Effenberger, Frank; Liu, Xiang

    2015-05-18

    We propose a power-efficient method for transmitting multiple frequency-division multiplexed (FDM) orthogonal frequency-division multiplexing (OFDM) signals in intensity-modulation direct-detection (IM-DD) optical systems. This method is based on quadratic soft clipping in combination with odd-only channel mapping. We show, both analytically and experimentally, that the proposed approach is capable of improving the power efficiency by about 3 dB as compared to conventional FDM OFDM signals under practical bias conditions, making it a viable solution in applications such as optical fiber-wireless integrated systems where both IM-DD optical transmission and OFDM signaling are important.

  12. ASBench: benchmarking sets for allosteric discovery.

    PubMed

    Huang, Wenkang; Wang, Guanqiao; Shen, Qiancheng; Liu, Xinyi; Lu, Shaoyong; Geng, Lv; Huang, Zhimin; Zhang, Jian

    2015-08-01

    Allostery allows for the fine-tuning of protein function. Targeting allosteric sites is gaining increasing recognition as a novel strategy in drug design. The key challenge in the discovery of allosteric sites has strongly motivated the development of computational methods and thus high-quality, publicly accessible standard data have become indispensable. Here, we report benchmarking data for experimentally determined allosteric sites through a complex process, including a 'Core set' with 235 unique allosteric sites and a 'Core-Diversity set' with 147 structurally diverse allosteric sites. These benchmarking sets can be exploited to develop efficient computational methods to predict unknown allosteric sites in proteins and reveal unique allosteric ligand-protein interactions to guide allosteric drug design.

  13. Tumor vasculature is regulated by FGF/FGFR signaling-mediated angiogenesis and bone marrow-derived cell recruitment: this mechanism is inhibited by SSR128129E, the first allosteric antagonist of FGFRs.

    PubMed

    Fons, Pierre; Gueguen-Dorbes, Geneviève; Herault, Jean-Pascal; Geronimi, Fabien; Tuyaret, Joël; Frédérique, Dol; Schaeffer, Paul; Volle-Challier, Cécile; Herbert, Jean-Marc; Bono, Françoise

    2015-01-01

    Tumor angiogenesis is accompanied by vasculogenesis, which is involved in the differentiation and mobilization of human bone marrow cells. In order to further characterize the role of vasculogenesis in the tumor growth process, the effects of FGF2 on the differentiation of human bone marrow AC133(+) cells (BM-AC133(+)) into vascular precursors were studied in vitro. FGF2, like VEGFA, induced progenitor cell differentiation into cell types with endothelial cell characteristics. SSR128129E, a newly discovered specific FGFR antagonist acting by allosteric interaction with FGFR, abrogated FGF2-induced endothelial cell differentiation, showing that FGFR signaling is essential during this process. To assess the involvement of the FGF/FRGR signaling in vivo, the pre-clinical model of Lewis lung carcinoma (LL2) in mice was used. Subcutaneous injection of LL2 cells into mice induced an increase of circulating EPCs from peripheral blood associated with tumor growth and an increase of intra-tumoral vascular index. Treatment with the FGFR antagonist SSR128129E strongly decreased LL2 tumor growth as well as the intra-tumoral vascular index (41% and 50% decrease vs. vehicle-treated mice respectively, P < 0.01). Interestingly, SSR128129E treatment significantly decreased the number of circulating EPCs from the peripheral blood (53% inhibition vs. vehicle-treated mice, P < 0.01). These results demonstrate for the first time that the blockade of the FGF/FGFR pathway by SSR128129E reduces EPC recruitment during angiogenesis-dependent tumor growth. In this context, circulating EPCs could be a reliable surrogate marker for tumor growth and angiogenic activity.

  14. Mitigation of nonlinear transmission effects for OFDM 16-QAM optical signal using adaptive modulation.

    PubMed

    Skidin, Anton S; Sidelnikov, Oleg S; Fedoruk, Mikhail P; Turitsyn, Sergei K

    2016-12-26

    The impact of the fiber Kerr effect on error statistics in the nonlinear (high power) transmission of the OFDM 16-QAM signal over a 2000 km EDFA-based link is examined. We observed and quantified the difference in the error statistics for constellation points located at three power-defined rings. Theoretical analysis of a trade-off between redundancy and error rate reduction using probabilistic coding of three constellation power rings decreasing the symbol-error rate of OFDM 16-QAM signal is presented. Based on this analysis, we propose to mitigate the nonlinear impairments using the adaptive modulation technique applied to the OFDM 16-QAM signal. We demonstrate through numerical modelling the system performance improvement by the adaptive modulation for the large number of OFDM subcarriers (more than 100). We also show that a similar technique can be applied to single carrier transmission.

  15. Multiple roles for mTOR signaling in both glutamatergic and GABAergic synaptic transmission

    PubMed Central

    Weston, Matthew C.; Chen, Hongmei; Swann, John W.

    2012-01-01

    Summary The mammalian target of rapamycin (mTOR) signaling pathway in neurons integrates a variety of extracellular signals to produce appropriate translational responses. mTOR signaling is hyperactive in neurological syndromes in both humans and mouse models that are characterized by epilepsy, autism and cognitive disturbances. In addition, rapamycin, a clinically important immunosuppressant, is a specific and potent inhibitor of mTOR signaling. While mTOR is known to regulate growth and synaptic plasticity of glutamatergic neurons, its effects on basic parameters of synaptic transmission are less well studied, and its role in regulating GABAergic transmission is unexplored. We therefore performed an electrophysiological and morphological comparison of glutamatergic and GABAergic neurons in which mTOR signaling was either increased by loss of the repressor Pten or decreased by treatment with rapamycin. We found that hyperactive mTOR signaling increased evoked synaptic responses in both glutamatergic and GABAergic neurons by approximately 50%, due to an increase in the number of synaptic vesicles available for release, the number of synapses formed and the miniature event size. Prolonged (72 hours) rapamycin treatment prevented these abnormalities and also decreased synaptic transmission in wild-type glutamatergic, but not GABAergic, neurons. Further analyses suggested that hyperactivation of the mTOR pathway also impairs presynaptic function, possibly by interfering with vesicle fusion. Despite this presynaptic impairment, the net effect of Pten loss is enhanced synaptic transmission in both GABAergic and glutamatergic neurons, which has numerous implications – depending on where in the brain mutations of an mTOR suppressor gene takes place during development. PMID:22895726

  16. Using optogenetics to interrogate the dynamic control of signal transmission by the Ras/Erk module.

    PubMed

    Toettcher, Jared E; Weiner, Orion D; Lim, Wendell A

    2013-12-05

    The complex, interconnected architecture of cell-signaling networks makes it challenging to disentangle how cells process extracellular information to make decisions. We have developed an optogenetic approach to selectively activate isolated intracellular signaling nodes with light and use this method to follow the flow of information from the signaling protein Ras. By measuring dose and frequency responses in single cells, we characterize the precision, timing, and efficiency with which signals are transmitted from Ras to Erk. Moreover, we elucidate how a single pathway can specify distinct physiological outcomes: by combining distinct temporal patterns of stimulation with proteomic profiling, we identify signaling programs that differentially respond to Ras dynamics, including a paracrine circuit that activates STAT3 only after persistent (>1 hr) Ras activation. Optogenetic stimulation provides a powerful tool for analyzing the intrinsic transmission properties of pathway modules and identifying how they dynamically encode distinct outcomes.

  17. Limb joint effects on signal transmission in capacitive coupled intra-body communication systems.

    PubMed

    Seyedi, MirHojjat; Lai, Daniel T H; Faulkner, Michael

    2012-01-01

    This paper contributes empirical measurements towards an understanding of signal attenuation in intra-body communication (IBC) systems due to limb posture effects. Recent studies have shown a degradation of transmission signals for IBC transmissions between limb segments, but these degradations have yet to be quantified with respect to relative limb position and within the transmission frequency range from 300 KHz to 200 MHz. We examine the impact of limb position specifically the effect of elbow joint flexion and extension into account using a portable vector network analyzer. The results presented indicate that the signal attenuation is larger in the case of extension, i.e., when the angle between forearm and upper arm increases. The minimum attenuation was 20.64 dB and 24.81 dB for the fix distance of 15 cm between transmitter and receiver electrodes and the joint angle of 45 and 180 degree respectively. It was found that attenuation decreased at an approximately linear rate over 300 KHz to 100 MHz and increased over the frequency range from 100 MHz to 200 MHz for the input signal frequency range from 300 KHz to 200 MHz. It was concluded that the minimum attenuation for the range of flexions and extensions occurred in the range 80-100 MHz. Future work will explore theoretical models to explain the observed results.

  18. Signal transmission through the CXC chemokine receptor 4 (CXCR4) transmembrane helices

    PubMed Central

    Wescott, Melanie P.; Kufareva, Irina; Paes, Cheryl; Goodman, Jason R.; Thaker, Yana; Puffer, Bridget A.; Berdougo, Eli; Rucker, Joseph B.; Handel, Tracy M.; Doranz, Benjamin J.

    2016-01-01

    The atomic-level mechanisms by which G protein-coupled receptors (GPCRs) transmit extracellular ligand binding events through their transmembrane helices to activate intracellular G proteins remain unclear. Using a comprehensive library of mutations covering all 352 residues of the GPCR CXC chemokine receptor 4 (CXCR4), we identified 41 amino acids that are required for signaling induced by the chemokine ligand CXCL12 (stromal cell-derived factor 1). CXCR4 variants with each of these mutations do not signal properly but remain folded, based on receptor surface trafficking, reactivity to conformationally sensitive monoclonal antibodies, and ligand binding. When visualized on the structure of CXCR4, the majority of these residues form a continuous intramolecular signaling chain through the transmembrane helices; this chain connects chemokine binding residues on the extracellular side of CXCR4 to G protein-coupling residues on its intracellular side. Integrated into a cohesive model of signal transmission, these CXCR4 residues cluster into five functional groups that mediate (i) chemokine engagement, (ii) signal initiation, (iii) signal propagation, (iv) microswitch activation, and (v) G protein coupling. Propagation of the signal passes through a “hydrophobic bridge” on helix VI that coordinates with nearly every known GPCR signaling motif. Our results agree with known conserved mechanisms of GPCR activation and significantly expand on understanding the structural principles of CXCR4 signaling. PMID:27543332

  19. Call transmission efficiency in native and invasive anurans: competing hypotheses of divergence in acoustic signals.

    PubMed

    Llusia, Diego; Gómez, Miguel; Penna, Mario; Márquez, Rafael

    2013-01-01

    Invasive species are a leading cause of the current biodiversity decline, and hence examining the major traits favouring invasion is a key and long-standing goal of invasion biology. Despite the prominent role of the advertisement calls in sexual selection and reproduction, very little attention has been paid to the features of acoustic communication of invasive species in nonindigenous habitats and their potential impacts on native species. Here we compare for the first time the transmission efficiency of the advertisement calls of native and invasive species, searching for competitive advantages for acoustic communication and reproduction of introduced taxa, and providing insights into competing hypotheses in evolutionary divergence of acoustic signals: acoustic adaptation vs. morphological constraints. Using sound propagation experiments, we measured the attenuation rates of pure tones (0.2-5 kHz) and playback calls (Lithobates catesbeianus and Pelophylax perezi) across four distances (1, 2, 4, and 8 m) and over two substrates (water and soil) in seven Iberian localities. All factors considered (signal type, distance, substrate, and locality) affected transmission efficiency of acoustic signals, which was maximized with lower frequency sounds, shorter distances, and over water surface. Despite being broadcast in nonindigenous habitats, the advertisement calls of invasive L. catesbeianus were propagated more efficiently than those of the native species, in both aquatic and terrestrial substrates, and in most of the study sites. This implies absence of optimal relationship between native environments and propagation of acoustic signals in anurans, in contrast to what predicted by the acoustic adaptation hypothesis, and it might render these vertebrates particularly vulnerable to intrusion of invasive species producing low frequency signals, such as L. catesbeianus. Our findings suggest that mechanisms optimizing sound transmission in native habitat can play a less

  20. Call Transmission Efficiency in Native and Invasive Anurans: Competing Hypotheses of Divergence in Acoustic Signals

    PubMed Central

    Llusia, Diego; Gómez, Miguel; Penna, Mario; Márquez, Rafael

    2013-01-01

    Invasive species are a leading cause of the current biodiversity decline, and hence examining the major traits favouring invasion is a key and long-standing goal of invasion biology. Despite the prominent role of the advertisement calls in sexual selection and reproduction, very little attention has been paid to the features of acoustic communication of invasive species in nonindigenous habitats and their potential impacts on native species. Here we compare for the first time the transmission efficiency of the advertisement calls of native and invasive species, searching for competitive advantages for acoustic communication and reproduction of introduced taxa, and providing insights into competing hypotheses in evolutionary divergence of acoustic signals: acoustic adaptation vs. morphological constraints. Using sound propagation experiments, we measured the attenuation rates of pure tones (0.2–5 kHz) and playback calls (Lithobates catesbeianus and Pelophylax perezi) across four distances (1, 2, 4, and 8 m) and over two substrates (water and soil) in seven Iberian localities. All factors considered (signal type, distance, substrate, and locality) affected transmission efficiency of acoustic signals, which was maximized with lower frequency sounds, shorter distances, and over water surface. Despite being broadcast in nonindigenous habitats, the advertisement calls of invasive L. catesbeianus were propagated more efficiently than those of the native species, in both aquatic and terrestrial substrates, and in most of the study sites. This implies absence of optimal relationship between native environments and propagation of acoustic signals in anurans, in contrast to what predicted by the acoustic adaptation hypothesis, and it might render these vertebrates particularly vulnerable to intrusion of invasive species producing low frequency signals, such as L. catesbeianus. Our findings suggest that mechanisms optimizing sound transmission in native habitat can play a

  1. SETI: The transmission rate of radio communication and the signal's detection

    NASA Astrophysics Data System (ADS)

    Fridman, P. A.

    2011-11-01

    The transmission rate of communication between radio telescopes on Earth and extraterrestrial intelligence (ETI) is here calculated up to distances of 1000 light years. Both phase-shift keying (PSK) and frequency-shift keying (FSK) modulation schemes are considered. It is shown that M-ary FSK is advantageous in terms of energy. Narrow-band pulses scattered over the spectrum sharing a common drift rate can be the probable signals of ETI. Modern SETI spectrum analyzers are well suited to searching for these types of signals. Such signals can be detected using the Hough transform which is a dedicated tool for detecting patterns in an image. The time-frequency plane representing the power output of the spectrum analyzer during the search for ETI gives an image from which the Hough transform (HT) can detect signal patterns with frequency drift.

  2. Transmission and reception of PDM dual-subcarrier coherent 16QAM-OFDM signals

    NASA Astrophysics Data System (ADS)

    Li, Fan; Zhang, Junwen; Yu, Jianjun; Li, Xinying

    2015-12-01

    In this paper, 16-Gbaud polarization-division-multiplexed (PDM) dual-subcarrier coherent optical orthogonal frequency division multiplexing (CO-OFDM) transmission and reception are successfully demonstrated without overhead. The in-phase and quadrature (I/Q) components of dual-subcarrier 16-ary quadrature amplitude modulation (QAM) OFDM signal are both seven-level signals in time domain, and thus can be equalized like a 49 QAM signal in time domain with cascaded multi-modulus algorithm (CMMA) equalization method. The experimental results show that there is no power penalty observed between optical back to back (OBTB) and after 80-km single-mode fiber-28 (SMF-28) with time domain CMMA equalization method. A 0.4 dB optical signal to noise ratio (OSNR) penalty in OBTB is observed when the bandwidth of channel is set at 26 GHz at the BER of 2.0 × 10-2.

  3. Optical radio-photonic channel for transmission of a coherent narrowband analog signal

    NASA Astrophysics Data System (ADS)

    Zhuk, D. I.; Denisyuk, I. Yu.; Fokina, M. I.

    2015-10-01

    The channel of an optical transmission line of coherent narrowband analog signal consisting of a continuous-wave laser, an electro-optic modulator, and a vector phase rotator based on electrically controlled fiber-optical 1 × 2 splitter and fixed delay lines is analyzed. The scheme is constructed from commercially available components used in digital optical communication systems. The applicability of components for analog and small-signal circuits is determined. Variation of radio signal phase in the range from 0° to 170° for radio signal frequencies between 1 and 2 GHz is demonstrated experimentally. It is shown that phase variation is a linear function of frequency in this range.

  4. Photonics aided ultra-wideband W-band signal generation and air space transmission

    NASA Astrophysics Data System (ADS)

    Li, Xinying; Yu, Jianjun

    2016-02-01

    We achieve several field trial demonstrations of ultra-wideband W-band millimeter-wave (mm-wave) signal generation and its long-distance air space transmission based on some enabling technologies and advanced devices. First, we demonstrated photonics generation and up to 1.7-km wireless delivery of 20-Gb/s polarization division multiplexing quadrature phase shift keying (PDM-QPSK) signal at W-band, adopting both optical and antenna polarization multiplexing. Then, we demonstrated photonics generation and up to 300-m wireless delivery of 80-Gb/s PDM-QPSK signal at W-band, adopting both optical and antenna polarization multiplexing as well as multi-band multiplexing. We also demonstrated photonics generation and up to 100-m wireless delivery of 100-Gb/s QPSK signal at W-band, adopting antenna polarization multiplexing.

  5. Synchronization transmission of spatiotemporal chaotic signal in the uncertain time-varying network

    NASA Astrophysics Data System (ADS)

    Lü, Ling; Chen, Liansong; Han, Changhui; Ge, Lianjun; Gao, Liyu

    2017-02-01

    In this paper, a new method is presented for the synchronization transmission of spatiotemporal chaotic signal in the uncertain time-varying network. By designing a special function to construct the Lyapunov function of the network, it is sure that the uncertain time-varying network can effectively synchronize the spatiotemporal chaotic signal generated by the synchronization target. At the same time, we also design the identification laws of uncertain parameters and the adaptive laws of the time-varying coupling matrix elements. Especially in our work, the nodes of the uncertain time-varying network and the synchronization target are different. Obviously, this research has the reference value for the application fields.

  6. cAMP-Signalling Regulates Gametocyte-Infected Erythrocyte Deformability Required for Malaria Parasite Transmission

    PubMed Central

    Thompson, Eloise; Breil, Florence; Lorthiois, Audrey; Dupuy, Florian; Cummings, Ross; Duffier, Yoann; Corbett, Yolanda; Mercereau-Puijalon, Odile; Vernick, Kenneth; Taramelli, Donatella; Baker, David A.; Langsley, Gordon; Lavazec, Catherine

    2015-01-01

    Blocking Plasmodium falciparum transmission to mosquitoes has been designated a strategic objective in the global agenda of malaria elimination. Transmission is ensured by gametocyte-infected erythrocytes (GIE) that sequester in the bone marrow and at maturation are released into peripheral blood from where they are taken up during a mosquito blood meal. Release into the blood circulation is accompanied by an increase in GIE deformability that allows them to pass through the spleen. Here, we used a microsphere matrix to mimic splenic filtration and investigated the role of cAMP-signalling in regulating GIE deformability. We demonstrated that mature GIE deformability is dependent on reduced cAMP-signalling and on increased phosphodiesterase expression in stage V gametocytes, and that parasite cAMP-dependent kinase activity contributes to the stiffness of immature gametocytes. Importantly, pharmacological agents that raise cAMP levels in transmissible stage V gametocytes render them less deformable and hence less likely to circulate through the spleen. Therefore, phosphodiesterase inhibitors that raise cAMP levels in P. falciparum infected erythrocytes, such as sildenafil, represent new candidate drugs to block transmission of malaria parasites. PMID:25951195

  7. Signal transmission techniques for large-scale nuclear fuel reprocessing applications

    SciTech Connect

    Herndon, J.N.; Bible, D.W.

    1985-01-01

    The RCE is currently developing a prototypic microwave-based signal transmission system for reprocessing cell applications. This system, being developed for use in the Advanced Integrated Maintenance System (AIMS), will operate in the 10-GHz frequency range. Provisions are being made for five real-time video channels, three bidirectional data channels at one megabaud data rate each, and two audio channels. The basic utility of the concept has been proven in a laboratory demonstration using gallium arsenide gunn diode transmitter/receivers with horn antennas. Unidirectional transmission of one real-time video channel over a distance of 200 ft was demonstrated. No evidence of multipath interference was detected even when the transmission path was surrounded by metallic reflectors. The microwave signal transmission system for the AIMS application is in final design. Fabrication in the ORNL instrument shops will begin in October 1985, and the system should be operational in the Maintenance Systems Test Area (MSTA) at ORNL in the latter half of 1986.

  8. Allosteric Dynamic Control of Binding

    PubMed Central

    Sumbul, Fidan; Acuner-Ozbabacan, Saliha Ece; Haliloglu, Turkan

    2015-01-01

    Proteins have a highly dynamic nature and there is a complex interrelation between their structural dynamics and binding behavior. By assuming various conformational ensembles, they perform both local and global fluctuations to interact with other proteins in a dynamic infrastructure adapted to functional motion. Here, we show that there is a significant association between allosteric mutations, which lead to high-binding-affinity changes, and the hinge positions of global modes, as revealed by a large-scale statistical analysis of data in the Structural Kinetic and Energetic Database of Mutant Protein Interactions (SKEMPI). We further examined the mechanism of allosteric dynamics by conducting studies on human growth hormone (hGH) and pyrin domain (PYD), and the results show how mutations at the hinge regions could allosterically affect the binding-site dynamics or induce alternative binding modes by modifying the ensemble of accessible conformations. The long-range dissemination of perturbations in local chemistry or physical interactions through an impact on global dynamics can restore the allosteric dynamics. Our findings suggest a mechanism for the coupling of structural dynamics to the modulation of protein interactions, which remains a critical phenomenon in understanding the effect of mutations that lead to functional changes in proteins. PMID:26338442

  9. Allosteric regulation of phenylalanine hydroxylase.

    PubMed

    Fitzpatrick, Paul F

    2012-03-15

    The liver enzyme phenylalanine hydroxylase is responsible for conversion of excess phenylalanine in the diet to tyrosine. Phenylalanine hydroxylase is activated by phenylalanine; this activation is inhibited by the physiological reducing substrate tetrahydrobiopterin. Phosphorylation of Ser16 lowers the concentration of phenylalanine for activation. This review discusses the present understanding of the molecular details of the allosteric regulation of the enzyme.

  10. [Neuronal mechanisms of motor signal transmission in thalamic Voi nucleus in spasmodic torticollis patients].

    PubMed

    Sedov, A S; Raeva, S N; Pavlenko, V B

    2014-01-01

    Neural mechanisms of motor signal transmission in ventrooral (Voi) nucleus of motor thalamus during the realization-of voluntary and involuntary abnormal (dystonic) movements in patients with spasmodic torticollis were investigated by means of microelectrode technique. The high reactivity of the cellular Voi elements to various functional (mainly motor) tests was proved. Analysis of neuronal activity showed: (1) the difference of neural mechanisms of motor signal transmission in the realization of voluntary movement with and without the involvement of the pathological axial neck muscles, as well as passive and abnormal involuntary dystonic movements; (2) significance of sensory component in the mechanisms of sensorimotor interactions during realization of voluntary and involuntary dystonic head and neck movements, causing the activation of the axial neck muscles; (3) important role of the rhythmic and synchronized neuronal activity in motor signal transmission during the realization of active and passive movements. Participation of Voi nucleus in pathological mechanisms of spasmodic torticollis was shown. The data obtained can be used for identificatiori of Voi thalamic nucleus during stereotactic neurosurgical operations in patients with spasmodic torticollis for selection the optimum destruction (stimulation) target and reduction of postoperative effects.

  11. Differences in Allosteric Communication Pipelines in the Inactive and Active States of a GPCR

    PubMed Central

    Bhattacharya, Supriyo; Vaidehi, Nagarajan

    2014-01-01

    G-protein-coupled receptors (GPCRs) are membrane proteins that allosterically transduce the signal of ligand binding in the extracellular (EC) domain to couple to proteins in the intracellular (IC) domain. However, the complete pathway of allosteric communication from the EC to the IC domain, including the role of individual amino acids in the pathway is not known. Using the correlation in torsion angle movements calculated from microseconds-long molecular-dynamics simulations, we elucidated the allosteric pathways in three different conformational states of β2-adrenergic receptor (β2AR): 1), the inverse-agonist-bound inactive state; 2), the agonist-bound intermediate state; and (3), the agonist- and G-protein-bound fully active state. The inactive state is less dynamic compared with the intermediate and active states, showing dense clusters of allosteric pathways (allosteric pipelines) connecting the EC with the IC domain. The allosteric pipelines from the EC domain to the IC domain are weakened in the intermediate state, thus decoupling the EC domain from the IC domain and making the receptor more dynamic compared with the other states. Also, the orthosteric ligand-binding site becomes the initiator region for allosteric communication in the intermediate state. This finding agrees with the paradigm that the nature of the agonist governs the specific signaling state of the receptor. These results provide an understanding of the mechanism of allosteric communication in class A GPCRs. In addition, our analysis shows that mutations that affect the ligand efficacy, but not the binding affinity, are located in the allosteric pipelines. This clarifies the role of such mutations, which has hitherto been unexplained. PMID:25028884

  12. Enhanced GABA Transmission Drives Bradykinesia Following Loss of Dopamine D2 Receptor Signaling.

    PubMed

    Lemos, Julia C; Friend, Danielle M; Kaplan, Alanna R; Shin, Jung Hoon; Rubinstein, Marcelo; Kravitz, Alexxai V; Alvarez, Veronica A

    2016-05-18

    Bradykinesia is a prominent phenotype of Parkinson's disease, depression, and other neurological conditions. Disruption of dopamine (DA) transmission plays an important role, but progress in understanding the exact mechanisms driving slowness of movement has been impeded due to the heterogeneity of DA receptor distribution on multiple cell types within the striatum. Here we show that selective deletion of DA D2 receptors (D2Rs) from indirect-pathway medium spiny neurons (iMSNs) is sufficient to impair locomotor activity, phenocopying DA depletion models of Parkinson's disease, despite this mouse model having intact DA transmission. There was a robust enhancement of GABAergic transmission and a reduction of in vivo firing in striatal and pallidal neurons. Mimicking D2R signaling in iMSNs with Gi-DREADDs restored the level of tonic GABAergic transmission and rescued the motor deficit. These findings indicate that DA, through D2R activation in iMSNs, regulates motor output by constraining the strength of GABAergic transmission.

  13. Optical-wireless-optical full link for polarization multiplexing quadrature amplitude/phase modulation signal transmission.

    PubMed

    Li, Xinying; Yu, Jianjun; Chi, Nan; Zhang, Junwen

    2013-11-15

    We propose and experimentally demonstrate an optical wireless integration system at the Q-band, in which up to 40 Gb/s polarization multiplexing multilevel quadrature amplitude/phase modulation (PM-QAM) signal can be first transmitted over 20 km single-mode fiber-28 (SMF-28), then delivered over a 2 m 2 × 2 multiple-input multiple-output wireless link, and finally transmitted over another 20 km SMF-28. The PM-QAM modulated wireless millimeter-wave (mm-wave) signal at 40 GHz is generated based on the remote heterodyning technique, and demodulated by the radio-frequency transparent photonic technique based on homodyne coherent detection and baseband digital signal processing. The classic constant modulus algorithm equalization is used at the receiver to realize polarization demultiplexing of the PM-QAM signal. For the first time, to the best of our knowledge, we realize the conversion of the PM-QAM modulated wireless mm-wave signal to the optical signal as well as 20 km fiber transmission of the converted optical signal.

  14. Allosteric signaling through an mGlu2 and 5-HT2A heteromeric receptor complex and its potential contribution to schizophrenia

    PubMed Central

    Moreno, José L.; Miranda-Azpiazu, Patricia; García-Bea, Aintzane; Younkin, Jason; Cui, Meng; Kozlenkov, Alexey; Ben-Ezra, Ariel; Voloudakis, Georgios; Fakira, Amanda K.; Baki, Lia; Ge, Yongchao; Georgakopoulos, Anastasios; Morón, José A.; Milligan, Graeme; López-Giménez, Juan F.; Robakis, Nikolaos K.; Logothetis, Diomedes E.; Meana, J. Javier; González-Maeso, Javier

    2016-01-01

    Heterotrimeric guanine nucleotide–binding protein (G protein)–coupled receptors (GPCRs) can form multiprotein complexes (heteromers), which can alter the pharmacology and functions of the constituent receptors. Previous findings demonstrated that the Gq/11-coupled serotonin 5-HT2A receptor and the Gi/o-coupled metabotropic glutamate 2 (mGlu2) receptor—GPCRs that are involved in signaling alterations associated with psychosis—assemble into a heteromeric complex in the mammalian brain. In single-cell experiments with various mutant versions of the mGlu2 receptor, we showed that stimulation of cells expressing mGlu2–5-HT2A heteromers with an mGlu2 agonist led to activation of Gq/11 proteins by the 5-HT2A receptors. For this crosstalk to occur, one of the mGlu2 subunits had to couple to Gi/o proteins, and we determined the relative location of the Gi/o-contacting subunit within the mGlu2 homodimer of the heteromeric complex. Additionally, mGlu2-dependent activation of Gq/11, but not Gi/o, was reduced in the frontal cortex of 5-HT2A knockout mice and was reduced in the frontal cortex of postmortem brains from schizophrenic patients. These findings offer structural insights into this important target in molecular psychiatry. PMID:26758213

  15. The Mechanism of Allosteric Inhibition of Protein Tyrosine Phosphatase 1B

    PubMed Central

    Lu, Shaoyong; Huang, Wenkang; Geng, Lv; Shen, Qiancheng; Zhang, Jian

    2014-01-01

    As the prototypical member of the PTP family, protein tyrosine phosphatase 1B (PTP1B) is an attractive target for therapeutic interventions in type 2 diabetes. The extremely conserved catalytic site of PTP1B renders the design of selective PTP1B inhibitors intractable. Although discovered allosteric inhibitors containing a benzofuran sulfonamide scaffold offer fascinating opportunities to overcome selectivity issues, the allosteric inhibitory mechanism of PTP1B has remained elusive. Here, molecular dynamics (MD) simulations, coupled with a dynamic weighted community analysis, were performed to unveil the potential allosteric signal propagation pathway from the allosteric site to the catalytic site in PTP1B. This result revealed that the allosteric inhibitor compound-3 induces a conformational rearrangement in helix α7, disrupting the triangular interaction among helix α7, helix α3, and loop11. Helix α7 then produces a force, pulling helix α3 outward, and promotes Ser190 to interact with Tyr176. As a result, the deviation of Tyr176 abrogates the hydrophobic interactions with Trp179 and leads to the downward movement of the WPD loop, which forms an H-bond between Asp181 and Glu115. The formation of this H-bond constrains the WPD loop to its open conformation and thus inactivates PTP1B. The discovery of this allosteric mechanism provides an overall view of the regulation of PTP1B, which is an important insight for the design of potent allosteric PTP1B inhibitors. PMID:24831294

  16. Analysis of transmission loss, signal gain, and coherence in shallow water

    NASA Astrophysics Data System (ADS)

    Rozenfeld, Ilya

    2001-08-01

    Experiments in the Strait of Korea were performed to study sound propagation in an oceanographically complex shallow water environment. First a geoacoustic model is developed based on narrowband transmission loss measurements and using estimated profiles and measured bathymetry. The comparisons between measured and calculated transmission loss are made through an effective attenuation coefficient, which measures the rate of change of mean transmission loss with range. Environmental model parameters are selected to achieve the best agreement in the comparisons. Nonlinear frequency dependence in the sediment attenuation profiles permits good agreement between the calculations and measured data. The developed geoacoustic model is then used to obtain predictions of broadband transmission loss and signal energy spread. Very good agreement between these predictions and corresponding independent measurements validates the geoacoustic model. Next signal gain measurements taken during the experiment are examined. Using the previously developed environmental profiles the signal gain is computed. The calculations are in agreement with measurements for shorter ranges. For longer ranges and higher frequencies disagreement is found between the calculations and measurements. The cause is random fluctuations in the signal induced by the random medium. These effects can be included into the signal gain through the coherence function. Using a previously developed theory preliminary calculations of coherence are made. By choosing physically reasonable parameters of the random fluctuations in the medium, close agreement with measurements is achieved. Next this theory is extended to include scattering from inhomogeneities with arbitrary correlation functions. This allows a treatment of random fluctuations described by physically based spectra. The correlation functions corresponding to these spectra for mechanisms such as internal waves, turbulence, and wind driven sea surface

  17. The effect of habitat acoustics on common marmoset vocal signal transmission.

    PubMed

    Morrill, Ryan J; Thomas, A Wren; Schiel, Nicola; Souto, Antonio; Miller, Cory T

    2013-09-01

    Noisy acoustic environments present several challenges for the evolution of acoustic communication systems. Among the most significant is the need to limit degradation of spectro-temporal signal structure in order to maintain communicative efficacy. This can be achieved by selecting for several potentially complementary processes. Selection can act on behavioral mechanisms permitting signalers to control the timing and occurrence of signal production to avoid acoustic interference. Likewise, the signal itself may be the target of selection, biasing the evolution of its structure to comprise acoustic features that avoid interference from ambient noise or degrade minimally in the habitat. Here, we address the latter topic for common marmoset (Callithrix jacchus) long-distance contact vocalizations, known as phee calls. Our aim was to test whether this vocalization is specifically adapted for transmission in a species-typical forest habitat, the Atlantic forests of northeastern Brazil. We combined seasonal analyses of ambient habitat acoustics with experiments in which pure tones, clicks, and vocalizations were broadcast and rerecorded at different distances to characterize signal degradation in the habitat. Ambient sound was analyzed from intervals throughout the day and over rainy and dry seasons, showing temporal regularities across varied timescales. Broadcast experiment results indicated that the tone and click stimuli showed the typically inverse relationship between frequency and signaling efficacy. Although marmoset phee calls degraded over distance with marked predictability compared with artificial sounds, they did not otherwise appear to be specially designed for increased transmission efficacy or minimal interference in this habitat. We discuss these data in the context of other similar studies and evidence of potential behavioral mechanisms for avoiding acoustic interference in order to maintain effective vocal communication in common marmosets.

  18. Echolocation signals and transmission beam pattern of a false killer whale (Pseudorca crassidens).

    PubMed

    Au, W W; Pawloski, J L; Nachtigall, P E; Blonz, M; Gisner, R C

    1995-07-01

    The echolocation transmission beam pattern of a false killer whale (Pseudorca crassidens) was measured in the vertical and horizontal planes. A vertical array of seven broadband miniature hydrophones was used to measure the beam pattern in the vertical plane and a horizontal array of the same hydrophones was used in the horizontal plane. The measurements were performed in the open waters of Kaneohe Bay, Oahu, Hawaii, while the whale performed a target discrimination task. Four types of signals, characterized by their frequency spectra, were measured. Type-1 signals had a single low-frequency peak at 40 +/- 9 kHz and a low-amplitude shoulder at high frequencies. Type-2 signals had a bimodal frequency characteristic with a primary peak at 46 +/- 7 kHz and a secondary peak at 88 +/- 13 kHz. Type-3 signals were also bimodal but with a primary peak at 100 +/- 7 kHz and a secondary peak at 49 +/- 9 kHz. Type-4 signals had a single high-frequency peak at 104 +/- 7 kHz. The center frequency of the signals were found to be linearly correlated to the peak-to-peak source level, increasing with increasing source level. The major axis of the vertical beam was directed slightly downward between 0 and -5 degrees, in contrast to the +5 to 10 degrees for Tursiops and Delphinapterus. The beam in the horizontal plane was directed forward between 0 degrees and -5 degrees.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Extracellular Loop 2 of the Free Fatty Acid Receptor 2 Mediates Allosterism of a Phenylacetamide Ago-Allosteric ModulatorS⃞

    PubMed Central

    Smith, Nicola J.; Ward, Richard J.; Stoddart, Leigh A.; Hudson, Brian D.; Kostenis, Evi; Ulven, Trond; Morris, Joanne C.; Tränkle, Christian; Tikhonova, Irina G.; Adams, David R.

    2011-01-01

    Allosteric agonists are powerful tools for exploring the pharmacology of closely related G protein-coupled receptors that have nonselective endogenous ligands, such as the short chain fatty acids at free fatty acid receptors 2 and 3 (FFA2/GPR43 and FFA3/GPR41, respectively). We explored the molecular mechanisms mediating the activity of 4-chloro-α-(1-methylethyl)-N-2-thiazolylbenzeneacetamide (4-CMTB), a recently described phenylacetamide allosteric agonist and allosteric modulator of endogenous ligand function at human FFA2, by combining our previous knowledge of the orthosteric binding site with targeted examination of 4-CMTB structure-activity relationships and mutagenesis and chimeric receptor generation. Here we show that 4-CMTB is a selective agonist for FFA2 that binds to a site distinct from the orthosteric site of the receptor. Ligand structure-activity relationship studies indicated that the N-thiazolyl amide is likely to provide hydrogen bond donor/acceptor interactions with the receptor. Substitution at Leu173 or the exchange of the entire extracellular loop 2 of FFA2 with that of FFA3 was sufficient to reduce or ablate, respectively, allosteric communication between the endogenous and allosteric agonists. Thus, we conclude that extracellular loop 2 of human FFA2 is required for transduction of cooperative signaling between the orthosteric and an as-yet-undefined allosteric binding site of the FFA2 receptor that is occupied by 4-CMTB. PMID:21498659

  20. Light-activated DNA binding in a designed allosteric protein

    SciTech Connect

    Strickland, Devin; Moffat, Keith; Sosnick, Tobin R.

    2008-09-03

    An understanding of how allostery, the conformational coupling of distant functional sites, arises in highly evolvable systems is of considerable interest in areas ranging from cell biology to protein design and signaling networks. We reasoned that the rigidity and defined geometry of an {alpha}-helical domain linker would make it effective as a conduit for allosteric signals. To test this idea, we rationally designed 12 fusions between the naturally photoactive LOV2 domain from Avena sativa phototropin 1 and the Escherichia coli trp repressor. When illuminated, one of the fusions selectively binds operator DNA and protects it from nuclease digestion. The ready success of our rational design strategy suggests that the helical 'allosteric lever arm' is a general scheme for coupling the function of two proteins.

  1. Terabit Nyquist PDM-32QAM signal transmission with training sequence based time domain channel estimation.

    PubMed

    Zhang, Fan; Wang, Dan; Ding, Rui; Chen, Zhangyuan

    2014-09-22

    We propose a time domain structure of channel estimation for coherent optical communication systems, which employs training sequence based equalizer and is transparent to arbitrary quadrature amplitude modulation (QAM) formats. Enabled with this methodology, 1.02 Tb/s polarization division multiplexed 32 QAM Nyquist pulse shaping signal with a net spectral efficiency of 7.46 b/s/Hz is transmitted over standard single-mode fiber link with Erbium-doped fiber amplifier only amplification. After 1190 km transmission, the average bit-error rate is lower than the 20% hard-decision forward error correction threshold of 1.5 × 10(-2). The transmission distance can be extended to 1428 km by employing intra-subchannel nonlinear compensation with the digital back-propagation method.

  2. Effective simulation method for parametric signal-noise interaction in transmission fibers.

    PubMed

    Vanin, Evgeny; Jacobsen, Gunnar; Berntson, Anders

    2006-08-01

    We propose a new method for effective numerical simulation of transmission system performance and study of correlated noise evolution along an optical fiber with nonlinear parametric interaction between the amplified spontaneous emission (ASE) and the modulated optical signal. The method is based on an evaluation of the noise covariance matrix by using full nonlinear Schrödinger equation (NLSE) and an analytical model for the optical receiver. Using extensive brute-force Monte Carlo simulation as a verification tool, we test the accuracy of the method and illustrate the analytical receiver model limitations in the case of moderate as well as substantial growth of non-Gaussian optical noise along the optical fiber transmission link.

  3. Multiplex transmission system for gate drive signals of inverter circuit using surface acoustic wave filters

    NASA Astrophysics Data System (ADS)

    Suzuki, Akifumi; Ueda, Kensuke; Goka, Shigeyoshi; Wada, Keiji; Kakio, Shoji

    2016-07-01

    We propose and fabricate a multiplexed transmission system based on frequency-division multiple access (FDMA) with surface acoustic wave (SAW) filters. SAW filters are suitable for use in wide-gap switching devices and multilevel inverters because of their capability to operate at high temperatures, good electrical isolation, low cost, and high reliability. Our proposed system reduces the number of electrical signal wires needed to control each switching device and eliminates the need for isolation circuits, simplifying the transmission system and gate drive circuits. We successfully controlled two switching devices with a single coaxial line and confirmed the operation of a single-phase half-bridge inverter at a supply voltage of 100 V, and the total delay time to control the switching devices was less than 2.5 µs. Our experimental results validated our proposed system.

  4. Corticomuscular Transmission of Tremor Signals by Propriospinal Neurons in Parkinson's Disease

    PubMed Central

    Hao, Manzhao; He, Xin; Xiao, Qin; Alstermark, Bror; Lan, Ning

    2013-01-01

    Cortical oscillatory signals of single and double tremor frequencies act together to cause tremor in the peripheral limbs of patients with Parkinson's disease (PD). But the corticospinal pathway that transmits the tremor signals has not been clarified, and how alternating bursts of antagonistic muscle activations are generated from the cortical oscillatory signals is not well understood. This paper investigates the plausible role of propriospinal neurons (PN) in C3–C4 in transmitting the cortical oscillatory signals to peripheral muscles. Kinematics data and surface electromyogram (EMG) of tremor in forearm were collected from PD patients. A PN network model was constructed based on known neurophysiological connections of PN. The cortical efferent signal of double tremor frequencies were integrated at the PN network, whose outputs drove the muscles of a virtual arm (VA) model to simulate tremor behaviors. The cortical efferent signal of single tremor frequency actuated muscle spindles. By comparing tremor data of PD patients and the results of model simulation, we examined two hypotheses regarding the corticospinal transmission of oscillatory signals in Parkinsonian tremor. Hypothesis I stated that the oscillatory cortical signals were transmitted via the mono-synaptic corticospinal pathways bypassing the PN network. The alternative hypothesis II stated that they were transmitted by way of PN multi-synaptic corticospinal pathway. Simulations indicated that without the PN network, the alternating burst patterns of antagonistic muscle EMGs could not be reliably generated, rejecting the first hypothesis. However, with the PN network, the alternating burst patterns of antagonist EMGs were naturally reproduced under all conditions of cortical oscillations. The results suggest that cortical commands of single and double tremor frequencies are further processed at PN to compute the alternating burst patterns in flexor and extensor muscles, and the neuromuscular dynamics

  5. Low power wireless ultra-wide band transmission of bio-signals

    NASA Astrophysics Data System (ADS)

    Gabrielli, A.; Bastianini, S.; Crepaldi, M.; D'Amen, G.; Demarchi, D.; Lax, I.; Motto Ros, P.; Zoccoli, G.

    2014-12-01

    The paper shows the design of microelectronic circuits composed of an oscillator, a modulator, a transmitter and an antenna. Prototype chips were recently fabricated and tested exploiting commercial 130 nm [1] and 180 nm [2,3] CMOS technologies. Detected signals have been measured using a commercial Ultra-Wide-Band amplifier connected to custom designed filters and a digital demodulator. Preliminary results are summarized along with some waveforms of the transmitted and received signals. A digital Synchronized On-Off Keying (S-OOK) was implemented to exploit the Ultra-Wide-Band transmission. In this way, each transmitted bit is coded with a S-OOK protocol. Wireless transmission capabilities of the system have been also evaluated within a one-meter distance. The chips fit a large variety of applications like spot radiation monitoring, punctual measurements of radiation in High-Energy Physics experiments or, since they have been characterized as low-power components, readout of the system for medical applications. These latter fields are those that we are investigating for in-vivo measurements on small animals. In more detail, if we refer to electromyographic, electrocardiographic or electroencephalographic signals [4], we need to handle very small signal amplitudes, of the order of tens of μV, overwhelmed with a much higher (white) noise. In these cases the front-end of the readout circuit requires a so-called amplifier for instrumentation, here not described, to interface with metal-plate sensor's outputs such those used for electrocardiograms, to normal range of amplitude signals of the order of 1 V. We are also studying these circuits, to be also designed on a microelectronic device, without adding further details since these components are technically well known in the literature [5,6]. The main aim of this research is hence integrating all the described electronic components into a very small, low-powered, microelectronic circuit fully compatible with in

  6. A Study of Mechanical Vibration Signal Transmission Using Position Modulated Optical Pulses

    NASA Astrophysics Data System (ADS)

    Yamashita, Ikuo; Oro, Kyoichi; Seikai, Shigeyuki

    A novel vibration sensing scheme using a technique of converting the vibration into position modulated optical pulses is described. The laser light whose wavelength changes at a cycle of several kHz is launched into a fiber and passes through an optical filter whose center wavelength is changed in proportion to the mechanical vibration with frequency lower than kHz. The output signal from the filter becomes time position modulated optical pulses because the laser light passes only when the both wavelengths coincide. The basic operation of the scheme is experimentally confirmed using a 5-km transmission line.

  7. Wnt signaling pathway improves central inhibitory synaptic transmission in a mouse model of Duchenne muscular dystrophy.

    PubMed

    Fuenzalida, Marco; Espinoza, Claudia; Pérez, Miguel Ángel; Tapia-Rojas, Cheril; Cuitino, Loreto; Brandan, Enrique; Inestrosa, Nibaldo C

    2016-02-01

    The dystrophin-associated glycoprotein complex (DGC) that connects the cytoskeleton, plasma membrane and the extracellular matrix has been related to the maintenance and stabilization of channels and synaptic receptors, which are both essential for synaptogenesis and synaptic transmission. The dystrophin-deficient (mdx) mouse model of Duchenne muscular dystrophy (DMD) exhibits a significant reduction in hippocampal GABA efficacy, which may underlie the altered synaptic function and abnormal hippocampal long-term plasticity exhibited by mdx mice. Emerging studies have implicated Wnt signaling in the modulation of synaptic efficacy, neuronal plasticity and cognitive function. We report here that the activation of the non-canonical Wnt-5a pathway and Andrographolide, improves hippocampal mdx GABAergic efficacy by increasing the number of inhibitory synapses and GABA(A) receptors or GABA release. These results indicate that Wnt signaling modulates GABA synaptic efficacy and could be a promising novel target for DMD cognitive therapy.

  8. The structure of Escherichia coli signal recognition particle revealed by scanning transmission electron microscopy.

    PubMed

    Mainprize, Iain L; Beniac, Daniel R; Falkovskaia, Elena; Cleverley, Robert M; Gierasch, Lila M; Ottensmeyer, F Peter; Andrews, David W

    2006-12-01

    Structural studies on various domains of the ribonucleoprotein signal recognition particle (SRP) have not converged on a single complete structure of bacterial SRP consistent with the biochemistry of the particle. We obtained a three-dimensional structure for Escherichia coli SRP by cryoscanning transmission electron microscopy and mapped the internal RNA by electron spectroscopic imaging. Crystallographic data were fit into the SRP reconstruction, and although the resulting model differed from previous models, they could be rationalized by movement through an interdomain linker of Ffh, the protein component of SRP. Fluorescence resonance energy transfer experiments determined interdomain distances that were consistent with our model of SRP. Docking our model onto the bacterial ribosome suggests a mechanism for signal recognition involving interdomain movement of Ffh into and out of the nascent chain exit site and suggests how SRP could interact and/or compete with the ribosome-bound chaperone, trigger factor, for a nascent chain during translation.

  9. Molecular dynamics approach to probe the allosteric inhibition of PTP1B by chlorogenic and cichoric acid.

    PubMed

    Baskaran, Sarath Kumar; Goswami, Nabajyoti; Selvaraj, Sudhagar; Muthusamy, Velusamy Shanmuganathan; Lakshmi, Baddireddi Subhadra

    2012-08-27

    Protein tyrosine phosphatase 1B (PTP1B), a major negative regulator of the insulin and leptin signaling pathway, is a potential target for therapeutic intervention against diabetes and obesity. The recent discovery of an allosteric site in PTP1B has created an alternate strategy in the development of PTP1B targeted therapy. The current study investigates the molecular interactions between the allosteric site of PTP1B with two caffeoyl derivatives, chlorogenic acid (CGA) and cichoric acid (CHA), using computational strategies. Molecular docking analysis with CGA and CHA at the allosteric site of PTP1B were performed and the resulting protein-ligand complexes used for molecular dynamics simulation studies for a time scale of 10 ns. Results show stable binding of CGA and CHA at the allosteric site of PTP1B. The flexibility of the WPD loop was observed to be constrained by CGA and CHA in the open (inactive), providing molecular mechanism of allosteric inhibition. The allosteric inhibition of CGA and CHA of PTP1B was shown to be favorable due to no restriction by the α-7 helix in the binding of CGA and CHA at the allosteric binding site. In conclusion, our results exhibit an inhibitory pattern of CGA and CHA against PTP1B through potent binding at the allosteric site.

  10. Experimental evaluation of prefiltering for 56 Gbaud DP-QPSK signal transmission in 75 GHz WDM grid

    NASA Astrophysics Data System (ADS)

    Borkowski, Robert; de Carvalho, Luis Henrique H.; Silva, Edson Porto da; Diniz, Júlio César M.; Zibar, Darko; de Oliveira, Júlio César R. F.; Tafur Monroy, Idelfonso

    2014-01-01

    We investigate optical prefiltering for 56 Gbaud (224 Gbit/s) electrical time-division multiplexed (ETDM) dual polarization (DP) quaternary phase shift keying (QPSK) transmission. Different transmitter-side optical filter shapes are tested and their bandwidths are varied. Comparison of studied filter shapes shows an advantage of a pre-emphasis filter. Subsequently, we perform a fiber transmission of the 56 Gbaud DP QPSK signal filtered with the 65 GHz pre-emphasis filter to fit the 75 GHz transmission grid. Bit error rate (BER) of the signal remains below forward error correction (FEC) limit after 300 km of fiber propagation.

  11. Monitoring of global acoustic transmissions: Signal processing and preliminary data analysis

    NASA Astrophysics Data System (ADS)

    Frogner, Gary R.

    1991-09-01

    A great deal of controversy exists concerning the possible global warming trend which may occur as a result of a documented increase in atmospheric greenhouse gasses. The 1991 Heard Island Feasibility Experiment tested the feasibility of using transmissions of acoustic energy through major ocean basins of the world to monitor spatially averaged global temperature trends. This thesis documents the Naval Postgraduate School's reception of the phase encoded signal transmitted from the Southern Indian Ocean, development of real-time signal processing software, and preliminary data analysis. Data, received from a 32-channel vertical array suspended in the deep sound channel off the coast of Monterey, CA, was processed using real-time capable software. Data processing to reduce noise, determine SNR, and remove the m-sequence coding was found to be quite sensitive to Doppler frequency shifts. Although the SNR of the raw data was only about -27.5 dB for individual hydrophones, the transmitted signal was detected in both the frequency and time domains. However, the maximum processed signal peak in the time domain had an SNR of only +9 dB which is insufficient for use in a long term global temperature monitoring project. The hydrophone provides inadequate arrival time resolution.

  12. Optical differential phase-shift keyed signal generation, transmission and detection

    NASA Astrophysics Data System (ADS)

    Lize, Yannick Keith

    When encoding information on an electromagnetic wave such as infrared light, to be transmitted through an optical fibre in telecommunication networks, any of the physical properties of light can be modulated. Light has a frequency, intensity, polarization and a phase. Until recently, optical communication systems strictly employed conventional intensity (IM) modulation signals in either non return-to-zero (NRZ) or return-to-zero (RZ) format. But a number of advanced optical modulation formats have attracted increasing attention in the last few years. One prime example is the phase-shift-keyed (PSK) family of formats which carry the information on the optical phase. Since absolute phase is not easily detected through coherent demodulation, differential encoding in which the phase of the preceding bit is used as a relative phase reference for demodulation has become a method of choice for phase modulated signals. The result in the differential-phase-shift-keyed (DPSK) formats, which carry the information in the difference in optical phase between successive bits. In this thesis by article, composed of six papers, we investigate the generation, transmission and demodulation of DPSK in optical fibre transmission systems. We propose a novel way to encode optical packets using DPSK in our investigation of the generation. We also investigate transmission effects monitoring using a novel partial-bit delay interferometer-assisted clock tone monitoring method for sensitive optical-signal-to-noise ratio (OSNR), chromatic dispersion and polarization mode dispersion monitoring. Then we look at the demodulation of DPSK, first investigating the reduced tolerances and power penalties of DPSK demodulation when more than one bit delay is used in the interferometer. We also propose an optical error correction method combining DPSK optical logic gates with electronic logic gates to improve receiver sensitivity and transmission impairment tolerances. Finally we redefine the previously

  13. Electrical signal transmission in a bone cell network: the influence of a discrete gap junction

    NASA Technical Reports Server (NTRS)

    Zhang, D.; Weinbaum, S.; Cowin, S. C.

    1998-01-01

    A refined electrical cable model is formulated to investigate the role of a discrete gap junction in the intracellular transmission of electrical signals in an electrically coupled system of osteocytes and osteoblasts in an osteon. The model also examines the influence of the ratio q between the membrane's electrical time constant and the characteristic time of pore fluid pressure, the circular, cylindrical geometry of the osteon, and key simplifying assumptions in our earlier continuous cable model (see Zhang, D., S. C. Cowin, and S. Weinbaum. Electrical signal transmission and gap junction regulation in a bone cell network: A cable model for an osteon. Ann. Biomed. Eng. 25:379-396, 1997). Using this refined model, it is shown that (1) the intracellular potential amplitude at the osteoblastic end of the osteonal cable retains the character of a combination of a low-pass and a high-pass filter as the corner frequency varies in the physiological range; (2) the presence of a discrete gap junction near a resting osteoblast can lead to significant modulation of the intracellular potential and current in the osteoblast for measured values of the gap junction coupling strength; and (3) the circular, cylindrical geometry of the osteon is well simulated by the beam analogy used in Zhang et al.

  14. M1 muscarinic allosteric modulators slow prion neurodegeneration and restore memory loss

    PubMed Central

    Bradley, Sophie J.; Bourgognon, Julie-Myrtille; Sanger, Helen E.; Verity, Nicholas; Mogg, Adrian J.; White, David J.; Butcher, Adrian J.; Moreno, Julie A.; Macedo-Hatch, Timothy; Edwards, Jennifer M.; Wess, Jurgen; Pawlak, Robert; Read, David J.; Sexton, Patrick M.; Broad, Lisa M.; Steinert, Joern R.; Mallucci, Giovanna R.; Felder, Christian C.

    2016-01-01

    The current frontline symptomatic treatment for Alzheimer’s disease (AD) is whole-body upregulation of cholinergic transmission via inhibition of acetylcholinesterase. This approach leads to profound dose-related adverse effects. An alternative strategy is to selectively target muscarinic acetylcholine receptors, particularly the M1 muscarinic acetylcholine receptor (M1 mAChR), which was previously shown to have procognitive activity. However, developing M1 mAChR–selective orthosteric ligands has proven challenging. Here, we have shown that mouse prion disease shows many of the hallmarks of human AD, including progressive terminal neurodegeneration and memory deficits due to a disruption of hippocampal cholinergic innervation. The fact that we also show that muscarinic signaling is maintained in both AD and mouse prion disease points to the latter as an excellent model for testing the efficacy of muscarinic pharmacological entities. The memory deficits we observed in mouse prion disease were completely restored by treatment with benzyl quinolone carboxylic acid (BQCA) and benzoquinazoline-12 (BQZ-12), two highly selective positive allosteric modulators (PAMs) of M1 mAChRs. Furthermore, prolonged exposure to BQCA markedly extended the lifespan of diseased mice. Thus, enhancing hippocampal muscarinic signaling using M1 mAChR PAMs restored memory loss and slowed the progression of mouse prion disease, indicating that this ligand type may have clinical benefit in diseases showing defective cholinergic transmission, such as AD. PMID:27991860

  15. M1 muscarinic allosteric modulators slow prion neurodegeneration and restore memory loss.

    PubMed

    Bradley, Sophie J; Bourgognon, Julie-Myrtille; Sanger, Helen E; Verity, Nicholas; Mogg, Adrian J; White, David J; Butcher, Adrian J; Moreno, Julie A; Molloy, Colin; Macedo-Hatch, Timothy; Edwards, Jennifer M; Wess, Jurgen; Pawlak, Robert; Read, David J; Sexton, Patrick M; Broad, Lisa M; Steinert, Joern R; Mallucci, Giovanna R; Christopoulos, Arthur; Felder, Christian C; Tobin, Andrew B

    2017-02-01

    The current frontline symptomatic treatment for Alzheimer's disease (AD) is whole-body upregulation of cholinergic transmission via inhibition of acetylcholinesterase. This approach leads to profound dose-related adverse effects. An alternative strategy is to selectively target muscarinic acetylcholine receptors, particularly the M1 muscarinic acetylcholine receptor (M1 mAChR), which was previously shown to have procognitive activity. However, developing M1 mAChR-selective orthosteric ligands has proven challenging. Here, we have shown that mouse prion disease shows many of the hallmarks of human AD, including progressive terminal neurodegeneration and memory deficits due to a disruption of hippocampal cholinergic innervation. The fact that we also show that muscarinic signaling is maintained in both AD and mouse prion disease points to the latter as an excellent model for testing the efficacy of muscarinic pharmacological entities. The memory deficits we observed in mouse prion disease were completely restored by treatment with benzyl quinolone carboxylic acid (BQCA) and benzoquinazoline-12 (BQZ-12), two highly selective positive allosteric modulators (PAMs) of M1 mAChRs. Furthermore, prolonged exposure to BQCA markedly extended the lifespan of diseased mice. Thus, enhancing hippocampal muscarinic signaling using M1 mAChR PAMs restored memory loss and slowed the progression of mouse prion disease, indicating that this ligand type may have clinical benefit in diseases showing defective cholinergic transmission, such as AD.

  16. Detectable signals of episodic risk effects on acute HIV transmission: strategies for analyzing transmission systems using genetic data.

    PubMed

    Alam, Shah Jamal; Zhang, Xinyu; Romero-Severson, Ethan Obie; Henry, Christopher; Zhong, Lin; Volz, Erik M; Brenner, Bluma G; Koopman, James S

    2013-03-01

    Episodic high-risk sexual behavior is common and can have a profound effect on HIV transmission. In a model of HIV transmission among men who have sex with men (MSM), changing the frequency, duration and contact rates of high-risk episodes can take endemic prevalence from zero to 50% and more than double transmissions during acute HIV infection (AHI). Undirected test and treat could be inefficient in the presence of strong episodic risk effects. Partner services approaches that use a variety of control options will be likely to have better effects under these conditions, but the question remains: What data will reveal if a population is experiencing episodic risk effects? HIV sequence data from Montreal reveals genetic clusters whose size distribution stabilizes over time and reflects the size distribution of acute infection outbreaks (AIOs). Surveillance provides complementary behavioral data. In order to use both types of data efficiently, it is essential to examine aspects of models that affect both the episodic risk effects and the shape of transmission trees. As a demonstration, we use a deterministic compartmental model of episodic risk to explore the determinants of the fraction of transmissions during acute HIV infection (AHI) at the endemic equilibrium. We use a corresponding individual-based model to observe AIO size distributions and patterns of transmission within AIO. Episodic risk parameters determining whether AHI transmission trees had longer chains, more clustered transmissions from single individuals, or different mixes of these were explored. Encouragingly for parameter estimation, AIO size distributions reflected the frequency of transmissions from acute infection across divergent parameter sets. Our results show that episodic risk dynamics influence both the size and duration of acute infection outbreaks, thus providing a possible link between genetic cluster size distributions and episodic risk dynamics.

  17. Detectable signals of episodic risk effects on acute HIV transmission: Strategies for analyzing transmission systems using genetic data

    PubMed Central

    Alam, Shah Jamal; Zhang, Xinyu; Romero-Severson, Ethan Obie; Henry, Christopher; Zhong, Lin; Volz, Erik M.; Brenner, Bluma G.; Koopman, James S.

    2013-01-01

    Episodic high-risk sexual behavior is common and can have a profound effect on HIV transmission. In a model of HIV transmission among men who have sex with men (MSM), changing the frequency, duration and contact rates of high-risk episodes can take endemic prevalence from zero to 50% and more than double transmissions during acute HIV infection (AHI). Undirected test and treat could be inefficient in the presence of strong episodic risk effects. Partner services approaches that use a variety of control options will be likely to have better effects under these conditions, but the question remains: What data will reveal if a population is experiencing episodic risk effects? HIV sequence data from Montreal reveals genetic clusters whose size distribution stabilizes over time and reflects the size distribution of acute infection outbreaks (AIOs). Surveillance provides complementary behavioral data. In order to use both types of data efficiently, it is essential to examine aspects of models that affect both the episodic risk effects and the shape of transmission trees. As a demonstration, we use a deterministic compartmental model of episodic risk to explore the determinants of the fraction of transmissions during acute HIV infection (AHI) at the endemic equilibrium. We use a corresponding individual-based model to observe AIO size distributions and patterns of transmission within AIO. Episodic risk parameters determining whether AHI transmission trees had longer chains, more clustered transmissions from single individuals, or different mixes of these were explored. Encouragingly for parameter estimation, AIO size distributions reflected the frequency of transmissions from acute infection across divergent parameter sets. Our results show that episodic risk dynamics influence both the size and duration of acute infection outbreaks, thus providing a possible link between genetic cluster size distributions and episodic risk dynamics. PMID:23438430

  18. CGP7930: a positive allosteric modulator of the GABAB receptor.

    PubMed

    Adams, C L; Lawrence, A J

    2007-01-01

    CGP7930 (3-(3',5'-Di-tert-butyl-4'-hydroxy)phenyl-2,2-dimethylpropanol) is a positive allosteric modulator of the metabotropic GABAB receptor. CGP7930 has been found to modulate the GABAB receptor in the open, or high affinity, state increasing agonist affinity for the receptor and signal transduction efficacy following agonist stimulation. The GABAB heteromeric subunit B2, involved in signal transduction but not ligand binding, seems to be the site of action of CGP7930 and similar allosteric modulators. When administered alone in naïve animals, CGP7930 acts as an anxiolytic in rodents without other overt behavioral effects and has also been demonstrated to reduce self-administration of nicotine, cocaine, or alcohol in rodents, suggesting that "fine tuning" of the GABAB receptor by positive allosteric modulators may be able to regulate abuse of these drugs. Baclofen, the GABAB agonist, is currently finding use in treating addiction and various other disorders, but this can result in off-target effects and tolerance. CGP7930 when co-administered with baclofen enhances its potency, which could in theory minimize deleterious effects. Further study of CGP7930 is required, but this compound, and others like it, holds potential in a clinical setting.

  19. [G-protein-coupled receptors targeting: the allosteric approach].

    PubMed

    Sebag, Julien A; Pantel, Jacques

    2012-10-01

    G-protein-coupled receptors (GPCR) are a major family of drug targets. Essentially all drugs targeting these receptors on the market compete with the endogenous ligand (agonists or antagonists) for binding the receptor. Recently, non-competitive compounds binding to distinct sites from the cognate ligand were documented in various classes of these receptors. These compounds, called allosteric modulators, generally endowed of a better selectivity are able to modulate specifically the endogenous signaling of the receptor. To better understand the promising potential of this class of GPCRs targeting compounds, this review highlights the properties of allosteric modulators, the strategies used to identify them and the challenges associated with the development of these compounds.

  20. Allosteric Modulation of Purine and Pyrimidine Receptors

    PubMed Central

    Jacobson, Kenneth A.; Gao, Zhan-Guo; Göblyös, Anikó; IJzerman, Adriaan P.

    2011-01-01

    Among the purine and pyrimidine receptors, the discovery of small molecular allosteric modulators has been most highly advanced for the A1 and A3 ARs. These AR modulators have allosteric effects that are structurally separated from the orthosteric effects in SAR studies. The benzoylthiophene derivatives tend to act as allosteric agonists, as well as selective positive allosteric modulators (PAMs) of the A1 AR. A 2-amino-3-aroylthiophene derivative T-62 has been under development as a PAM of the A1 AR for the treatment of chronic pain. Several structurally distinct classes of allosteric modulators of the human A3 AR have been reported: 3-(2-pyridinyl)isoquinolines, 2,4-disubstituted quinolines, 1H-imidazo-[4,5-c]quinolin-4-amines, endocannabinoid 2-arachidonylglycerol and the food dye Brilliant Black BN. Site-directed mutagenesis of A1 and A3 ARs has identified residues associated with the allosteric effect, distinct from those that affect orthosteric binding. A few small molecular allosteric modulators have been reported for several of the P2X ligand-gated ion channels and the G protein-coupled P2Y receptor nucleotides. Metal ion modulation of the P2X receptors has been extensively explored. The allosteric approach to modulation of purine and pyrimidine receptors looks promising for development of drugs that are event-specific and site-specific in action. PMID:21586360

  1. Structure-Based Statistical Mechanical Model Accounts for the Causality and Energetics of Allosteric Communication

    PubMed Central

    Guarnera, Enrico; Berezovsky, Igor N.

    2016-01-01

    Allostery is one of the pervasive mechanisms through which proteins in living systems carry out enzymatic activity, cell signaling, and metabolism control. Effective modeling of the protein function regulation requires a synthesis of the thermodynamic and structural views of allostery. We present here a structure-based statistical mechanical model of allostery, allowing one to observe causality of communication between regulatory and functional sites, and to estimate per residue free energy changes. Based on the consideration of ligand free and ligand bound systems in the context of a harmonic model, corresponding sets of characteristic normal modes are obtained and used as inputs for an allosteric potential. This potential quantifies the mean work exerted on a residue due to the local motion of its neighbors. Subsequently, in a statistical mechanical framework the entropic contribution to allosteric free energy of a residue is directly calculated from the comparison of conformational ensembles in the ligand free and ligand bound systems. As a result, this method provides a systematic approach for analyzing the energetics of allosteric communication based on a single structure. The feasibility of the approach was tested on a variety of allosteric proteins, heterogeneous in terms of size, topology and degree of oligomerization. The allosteric free energy calculations show the diversity of ways and complexity of scenarios existing in the phenomenology of allosteric causality and communication. The presented model is a step forward in developing the computational techniques aimed at detecting allosteric sites and obtaining the discriminative power between agonistic and antagonistic effectors, which are among the major goals in allosteric drug design. PMID:26939022

  2. Integration of radio-frequency transmission and radar in general software for multimodal battlefield signal modeling

    NASA Astrophysics Data System (ADS)

    Yamamoto, Kenneth K.; Reznicek, Nathan J.; Wilson, D. Keith

    2013-05-01

    The Environmental Awareness for Sensor and Emitter Employment (EASEE) software, being developed by the U. S. Army Engineer Research and Development Center (ERDC), provides a general platform for predicting sensor performance and optimizing sensor selection and placement in complex terrain and weather conditions. It incorporates an extensive library of target signatures, signal propagation models, and sensor systems. A flexible object-oriented design supports efficient integration and simulation of diverse signal modalities. This paper describes the integration of modeling capabilities for radio-frequency (RF) transmission and radar systems from the U. S. Navy Electromagnetic Propagation Integrated Resource Environment (EMPIRE), which contains nearly twenty different realistic RF propagation models. The integration utilizes an XML-based interface between EASEE and EMPIRE to set inputs for and run propagation models. To accommodate radars, fundamental improvements to the EASEE software architecture were made to support active-sensing scenarios with forward and backward propagation of the RF signals between the radar and target. Models for reflecting targets were defined to apply a target-specific, directionally dependent reflection coefficient (i.e., scattering cross section) to the incident wavefields.

  3. Power and signal transmission protocol for a contactless subdural spinal cord stimulation device.

    PubMed

    Song, Suk-Heung; Gillies, George T; Howard, Matthew A; Kuhnley, Brian; Utz, Marcel

    2013-02-01

    Wireless signal transmission will play a critical role in developing reliable subdural spinal cord stimulation systems. We have developed an approach to inductively coupling signals across the epidural spacing between the pial and epidural surfaces. The major design constraints include tolerance of coil misalignments from spinal cord geometries in addition to reasonable power efficiencies within the expected range of movement. The design of the primary side as a driving circuit is simplified by several turns of commercial magnetic wire, whereas the implanted secondary side is implemented in a micro-planar spiral coil tuned to a resonant frequency of 1.6 MHz. We present the results of wireless inductive coupling experiments that demonstrate the ability to transmit and receive a frequency modulated 1.6 MHz carrier signal between primary and secondary coil antennae scaled to 10 mm. Power delivery is in the range of 400 mW at a link efficiency of 32 % for strong coupling (coil separations of 0.5 mm ) and in the range of 70 mW at 4 % efficiency for weak coupling (coil separations of 10 mm).

  4. Transmission

    SciTech Connect

    Sugano, K.

    1988-12-27

    A transmission is described which consists of: an input shaft; an output shaft; a first planetary gear set including a first sun gear selectively connectable by a first clutch to the input shaft, a first carrier selectively connectable by a second clutch to the input shaft and a first ring gear connected to the output shaft. The first sun gear selectively held stationary by a first brake, the first carrier is allowed to rotate in the same forward direction as the input shaft when the second clutch is engaged, but prevented from rotating in a reverse direction opposite to the forward direction by a first one-way clutch, the first carrier being selectively held stationary by a second brake; a second planetary gear set including a second sun gear connected to the input shaft, a second carrier connected to the first ring gear and also the the output shaft, and a second ring gear.

  5. Allosteric binding sites on muscarinic acetylcholine receptors.

    PubMed

    Wess, Jürgen

    2005-12-01

    In this issue of Molecular Pharmacology, Tränkle et al. (p. 1597) present new findings regarding the existence of a second allosteric site on the M2 muscarinic acetylcholine receptor (M2 mAChR). The M2 mAChR is a prototypic class A G protein-coupled receptor (GPCR) that has proven to be a very useful model system to study the molecular mechanisms involved in the binding of allosteric GPCR ligands. Previous studies have identified several allosteric muscarinic ligands, including the acetylcholinesterase inhibitor tacrine and the bis-pyridinium derivative 4,4'-bis-[(2,6-dichloro-benzyloxy-imino)-methyl]-1,1'-propane-1,3-diyl-bis-pyridinium dibromide (Duo3), which, in contrast to conventional allosteric muscarinic ligands, display concentration-effect curves with slope factors >1. By analyzing the interactions of tacrine and Duo3 with other allosteric muscarinic agents predicted to bind to the previously identified ;common' allosteric binding site, Tränkle et al. provide evidence suggesting that two allosteric agents and one orthosteric ligand may be able to bind to the M2 mAChR simultaneously. Moreover, studies with mutant mAChRs indicated that the M2 receptor epitopes involved in the binding of tacrine and Duo3 may not be identical. Molecular modeling and ligand docking studies suggested that the additional allosteric site probably represents a subdomain of the receptor's allosteric binding cleft. Because allosteric binding sites have been found on many other GPCRs and drugs interacting with these sites are thought to have great therapeutic potential, the study by Tränkle et al. should be of considerable general interest.

  6. A Signal Transmission Technique for Stability Analysis of Multivariable Non-Linear Control Systems

    NASA Technical Reports Server (NTRS)

    Jackson, Mark; Zimpfer, Doug; Adams, Neil; Lindsey, K. L. (Technical Monitor)

    2000-01-01

    Among the difficulties associated with multivariable, non-linear control systems is the problem of assessing closed-loop stability. Of particular interest is the class of non-linear systems controlled with on/off actuators, such as spacecraft thrusters or electrical relays. With such systems, standard describing function techniques are typically too conservative, and time-domain simulation analysis is prohibitively extensive, This paper presents an open-loop analysis technique for this class of non-linear systems. The technique is centered around an innovative use of multivariable signal transmission theory to quantify the plant response to worst case control commands. The technique has been applied to assess stability of thruster controlled flexible space structures. Examples are provided for Space Shuttle attitude control with attached flexible payloads.

  7. High-frequency signal transmission through single-atom contacts of Au and Pt

    SciTech Connect

    Aoyama, Shodai; Kurokawa, Shu; Sakai, Akira

    2015-03-23

    Signal transmission through atom-sized contacts of Au and Pt has been studied at room temperature for frequencies from 9 kHz to 1 GHz and for conductances (1−10)G{sub 0} (G≡2e{sup 2}/h is the quantum unit of conductance). We measured the frequency spectrum of S parameter S{sub 21}=|S{sub 21}|e{sup iθ} and found θ∼0 up to 1 GHz for all contacts irrespective of their conductance. Our observations directly prove that the atom-sized contacts of Au and Pt, including their single-atom contacts, behave as a pure resistance in the RF regime.

  8. A Physiological Signal Transmission Model to be Used for Specific Diagnosis of Cochlear Impairments

    NASA Astrophysics Data System (ADS)

    Saremi, Amin; Stenfelt, Stefan

    2011-11-01

    Many of the sophisticated characteristics of human auditory system are attributed to cochlea. Also, most of patients with a hearing loss suffer from impairments that originate from cochlea (sensorineural). Despite this, today's clinical diagnosis methods do not probe the specific origins of such cochlear lesions. The aim of this research is to introduce a physiological signal transmission model to be clinically used as a tool for diagnosis of cochlear losses. This model enables simulation of different bio-mechano-electrical processes which occur in the auditory organ of Corti inside the cochlea. What makes this model different from many available computational models is its loyalty to physiology since the ultimate goal is to model each single physiological phenomenon. This includes passive BM vibration, outer hair cells' performances such as nonlinear mechanoelectrical transduction (MET), active amplifications by somatic motor, as well as vibration to neural conversion at the inner hair cells.

  9. The transmission of the NAO signal to alpine lakes in the Iberian Peninsula

    NASA Astrophysics Data System (ADS)

    Sánchez, Guiomar; Hernández, Armand; Toro, Manuel; Granados, Ignacio; Sigró, Javier; Pla-Rabes, Sergi; Trigo, Ricardo; Jesús Rubio, María; Giralt, Santiago

    2014-05-01

    The North Atlantic Oscillation (NAO) is one of the main climate circulation patterns ruling winter rainfall and temperature in western Europe. In particular, the NAO pattern controls to a large extent the seasonal and inter-annual precipitation variability in the Iberian Peninsula (IP). Alpine lake ecosystems can be excellent records of NAO influence. They have been shown to respond significantly to local and regional climate variability dominated by large-scale climatic fluctuations, including the NAO. Physical lake parameters seem to reflect these meteorological forcing more immediately and sensitively than other lacustrine ones (i.e biological parameters). Specifically, ice phenology has become one of the most valuable indicators of NAO winter influence. Many studies carried out in lakes located in Northern Hemisphere have in common to find this transmission through air temperature. In addition, only few works have found a significant relationship between NAO signal and other climate variables, such as precipitation or snow. Conversely, to the best of our knowledge this kind of assessments have not been performed yet in Southern Europe. Two alpine lakes, with a glacial origin and located in the Spanish Central Range (IP) have been selected to perform a conceptual model of the transmission of NAO signal to lakes: Cimera (dimictic at 2140 m a.s.l., 384 m long, 177 m wide and 9.4 m deep) and Peñalara (monomictic at 2017 m a.s.l., 115 m long, 71.5 m wide and 4.8 m deep). This conceptual model has been built using Pearson's r correlation coefficients between winter season (December-March) data sets of NAO index, local meteorology (precipitation, temperature and snow days) and limnology (ice phenology records and lake water surface temperatures) available for the period 1993-2011 in Lake Peñalara and for the period 2007-2013 in Lake Cimera. The conceptual model results suggest that NAO winter signal is mainly reflected in ice phenology by air temperature but also by

  10. Transmission of Predictable Sensory Signals to the Cerebellum via Climbing Fiber Pathways Is Gated during Exploratory Behavior

    PubMed Central

    Lawrenson, Charlotte L.; Watson, Thomas C.

    2016-01-01

    Pathways arising from the periphery that target the inferior olive [spino-olivocerebellar pathways (SOCPs)] are a vital source of information to the cerebellum and are modulated (gated) during active movements. This limits their ability to forward signals to climbing fibers in the cerebellar cortex. We tested the hypothesis that the temporal pattern of gating is related to the predictability of a sensory signal. Low-intensity electrical stimulation of the ipsilateral hindlimb in awake rats evoked field potentials in the C1 zone in the copula pyramidis of the cerebellar cortex. Responses had an onset latency of 12.5 ± 0.3 ms and were either short or long duration (8.7 ± 0.1 vs 31.2 ± 0.3 ms, respectively). Both types of response were shown to be mainly climbing fiber in origin and therefore evoked by transmission in hindlimb SOCPs. Changes in response size (area of field, millivolts per millisecond) were used to monitor differences in transmission during rest and three phases of rearing: phase 1, rearing up; phase 2, upright; and phase 3, rearing down. Responses evoked during phase 2 were similar in size to rest but were smaller during phases 1 and 3, i.e., transmission was reduced during active movement when self-generated (predictable) sensory signals from the hindlimbs are likely to occur. To test whether the pattern of gating was related to the predictability of the sensory signal, some animals received the hindlimb stimulation only during phase 2. Over ∼10 d, the responses became progressively smaller in size, consistent with gating-out transmission of predictable sensory signals relayed via SOCPs. SIGNIFICANCE STATEMENT A major route for peripheral information to gain access to the cerebellum is via ascending climbing fiber pathways. During active movements, gating of transmission in these pathways controls when climbing fiber signals can modify cerebellar activity. We investigated this phenomenon in rats during their exploratory behavior of rearing

  11. Inhibitory Mechanism of an Allosteric Antibody Targeting the Glucagon Receptor*

    PubMed Central

    Mukund, Susmith; Shang, Yonglei; Clarke, Holly J.; Madjidi, Azadeh; Corn, Jacob E.; Kates, Lance; Kolumam, Ganesh; Chiang, Vicky; Luis, Elizabeth; Murray, Jeremy; Zhang, Yingnan; Hötzel, Isidro; Koth, Christopher M.; Allan, Bernard B.

    2013-01-01

    Elevated glucagon levels and increased hepatic glucagon receptor (GCGR) signaling contribute to hyperglycemia in type 2 diabetes. We have identified a monoclonal antibody that inhibits GCGR, a class B G-protein coupled receptor (GPCR), through a unique allosteric mechanism. Receptor inhibition is mediated by the binding of this antibody to two distinct sites that lie outside of the glucagon binding cleft. One site consists of a patch of residues that are surface-exposed on the face of the extracellular domain (ECD) opposite the ligand-binding cleft, whereas the second binding site consists of residues in the αA helix of the ECD. A docking model suggests that the antibody does not occlude the ligand-binding cleft. We solved the crystal structure of GCGR ECD containing a naturally occurring G40S mutation and found a shift in the register of the αA helix that prevents antibody binding. We also found that alterations in the αA helix impact the normal function of GCGR. We present a model for the allosteric inhibition of GCGR by a monoclonal antibody that may form the basis for the development of allosteric modulators for the treatment of diabetes and other class B GPCR-related diseases. PMID:24189067

  12. Allosteric indicator displacement enzyme assay for a cyanogenic glycoside.

    PubMed

    Jose, D Amilan; Elstner, Martin; Schiller, Alexander

    2013-10-18

    Indicator displacement assays (IDAs) represent an elegant approach in supramolecular analytical chemistry. Herein, we report a chemical biosensor for the selective detection of the cyanogenic glycoside amygdalin in aqueous solution. The hybrid sensor consists of the enzyme β-glucosidase and a boronic acid appended viologen together with a fluorescent reporter dye. β-Glucosidase degrades the cyanogenic glycoside amygdalin into hydrogen cyanide, glucose, and benzaldehyde. Only the released cyanide binds at the allosteric site of the receptor (boronic acid) thereby inducing changes in the affinity of a formerly bound fluorescent indicator dye at the other side of the receptor. Thus, the sensing probe performs as allosteric indicator displacement assay (AIDA) for cyanide in water. Interference studies with inorganic anions and glucose revealed that cyanide is solely responsible for the change in the fluorescent signal. DFT calculations on a model compound revealed a 1:1 binding ratio of the boronic acid and cyanide ion. The fluorescent enzyme assay for β-glucosidase uses amygdalin as natural substrate and allows measuring Michaelis-Menten kinetics in microtiter plates. The allosteric indicator displacement assay (AIDA) probe can also be used to detect cyanide traces in commercial amygdalin samples.

  13. Conformationally Selective RNA Aptamers Allosterically Modulate the β2-Adrenoceptor

    PubMed Central

    Kahsai, Alem W.; Wisler, James W.; Lee, Jungmin; Ahn, Seungkirl; Cahill, Thomas J.; Dennison, S. Moses; Staus, Dean P.; Thomsen, Alex R. B.; Anasti, Kara M.; Pani, Biswaranjan; Wingler, Laura M.; Desai, Hemant; Bompiani, Kristin M.; Strachan, Ryan T.; Qin, Xiaoxia; Alam, S. Munir; Sullenger, Bruce A.; Lefkowitz, Robert J.

    2016-01-01

    G-protein-coupled receptor (GPCR) ligands function by stabilizing multiple, functionally distinct receptor conformations. This property underlies how “biased agonists” activate specific subsets of a given receptor’s signaling profile. However, stabilization of distinct active GPCR conformations to enable structural characterization of mechanisms underlying GPCR activation remains difficult. These challenges have accentuated the need for receptor tools that allosterically stabilize and regulate receptor function via unique, previously unappreciated mechanisms. Here, utilizing a highly diverse RNA library combined with advanced selection strategies involving state-of-the-art next-generation sequencing and bioinformatics analyses, we identify RNA aptamers that bind a prototypical GPCR, β2-adrenoceptor (β2AR). Using biochemical, pharmacological, and biophysical approaches, we demonstrate that these aptamers bind with nanomolar affinity at defined surfaces of the receptor, allosterically stabilizing active, inactive, and ligand-specific receptor conformations. The discovery of RNA aptamers as allosteric GPCR modulators significantly expands the diversity of ligands available to study the structural and functional regulation of GPCRs. PMID:27398998

  14. Inhibitory mechanism of an allosteric antibody targeting the glucagon receptor.

    PubMed

    Mukund, Susmith; Shang, Yonglei; Clarke, Holly J; Madjidi, Azadeh; Corn, Jacob E; Kates, Lance; Kolumam, Ganesh; Chiang, Vicky; Luis, Elizabeth; Murray, Jeremy; Zhang, Yingnan; Hötzel, Isidro; Koth, Christopher M; Allan, Bernard B

    2013-12-13

    Elevated glucagon levels and increased hepatic glucagon receptor (GCGR) signaling contribute to hyperglycemia in type 2 diabetes. We have identified a monoclonal antibody that inhibits GCGR, a class B G-protein coupled receptor (GPCR), through a unique allosteric mechanism. Receptor inhibition is mediated by the binding of this antibody to two distinct sites that lie outside of the glucagon binding cleft. One site consists of a patch of residues that are surface-exposed on the face of the extracellular domain (ECD) opposite the ligand-binding cleft, whereas the second binding site consists of residues in the αA helix of the ECD. A docking model suggests that the antibody does not occlude the ligand-binding cleft. We solved the crystal structure of GCGR ECD containing a naturally occurring G40S mutation and found a shift in the register of the αA helix that prevents antibody binding. We also found that alterations in the αA helix impact the normal function of GCGR. We present a model for the allosteric inhibition of GCGR by a monoclonal antibody that may form the basis for the development of allosteric modulators for the treatment of diabetes and other class B GPCR-related diseases.

  15. Transmission of 112 Gb/s PM-QPSK signals over up to 635 km of multimode optical fiber.

    PubMed

    Downie, John D; Hurley, Jason E; Kuksenkov, Dmitri V; Lynn, Christopher M; Korolev, Andrey E; Nazarov, Vladimir N

    2011-12-12

    We investigate transmission of 112 Gb/s PM-QPSK signals over 50 μm core diameter OM3 multimode fiber using the center launch approach. We demonstrate successful transmission of 16 DWDM channels over a distance of 635 km for a capacity-distance product of 1016 Tb/s-km. The limiting impairment appears due to mode coupling and multipath interference effects.

  16. Pin1-dependent signalling negatively affects GABAergic transmission by modulating neuroligin2/gephyrin interaction

    PubMed Central

    Antonelli, Roberta; Pizzarelli, Rocco; Pedroni, Andrea; Fritschy, Jean-Marc; Del Sal, Giannino; Cherubini, Enrico; Zacchi, Paola

    2014-01-01

    The cell adhesion molecule Neuroligin2 (NL2) is localized selectively at GABAergic synapses, where it interacts with the scaffolding protein gephyrin in the post-synaptic density. However, the role of this interaction for formation and plasticity of GABAergic synapses is unclear. Here, we demonstrate that endogenous NL2 undergoes proline-directed phosphorylation at its unique S714-P consensus site, leading to the recruitment of the peptidyl-prolyl cis–trans isomerase Pin1. This signalling cascade negatively regulates NL2’s ability to interact with gephyrin at GABAergic post-synaptic sites. As a consequence, enhanced accumulation of NL2, gephyrin and GABAA receptors was detected at GABAergic synapses in the hippocampus of Pin1-knockout mice (Pin1−/−) associated with an increase in amplitude of spontaneous GABAA-mediated post-synaptic currents. Our results suggest that Pin1-dependent signalling represents a mechanism to modulate GABAergic transmission by regulating NL2/gephyrin interaction. PMID:25297980

  17. Dynamic Coupling and Allosteric Networks in the α Subunit of Heterotrimeric G Proteins*

    PubMed Central

    Yao, Xin-Qiu; Malik, Rabia U.; Griggs, Nicholas W.; Skjærven, Lars; Traynor, John R.; Sivaramakrishnan, Sivaraj; Grant, Barry J.

    2016-01-01

    G protein α subunits cycle between active and inactive conformations to regulate a multitude of intracellular signaling cascades. Important structural transitions occurring during this cycle have been characterized from extensive crystallographic studies. However, the link between observed conformations and the allosteric regulation of binding events at distal sites critical for signaling through G proteins remain unclear. Here we describe molecular dynamics simulations, bioinformatics analysis, and experimental mutagenesis that identifies residues involved in mediating the allosteric coupling of receptor, nucleotide, and helical domain interfaces of Gαi. Most notably, we predict and characterize novel allosteric decoupling mutants, which display enhanced helical domain opening, increased rates of nucleotide exchange, and constitutive activity in the absence of receptor activation. Collectively, our results provide a framework for explaining how binding events and mutations can alter internal dynamic couplings critical for G protein function. PMID:26703464

  18. A dynamically coupled allosteric network underlies binding cooperativity in Src kinase

    PubMed Central

    Foda, Zachariah H.; Shan, Yibing; Kim, Eric T.; Shaw, David E.; Seeliger, Markus A.

    2015-01-01

    Protein tyrosine kinases are attractive drug targets because many human diseases are associated with the deregulation of kinase activity. However, how the catalytic kinase domain integrates different signals and switches from an active to an inactive conformation remains incompletely understood. Here we identify an allosteric network of dynamically coupled amino acids in Src kinase that connects regulatory sites to the ATP- and substrate-binding sites. Surprisingly, reactants (ATP and peptide substrates) bind with negative cooperativity to Src kinase while products (ADP and phosphopeptide) bind with positive cooperativity. We confirm the molecular details of the signal relay through the allosteric network by biochemical studies. Experiments on two additional protein tyrosine kinases indicate that the allosteric network may be largely conserved among these enzymes. Our work provides new insights into the regulation of protein tyrosine kinases and establishes a potential conduit by which resistance mutations to ATP-competitive kinase inhibitors can affect their activity. PMID:25600932

  19. A dynamically coupled allosteric network underlies binding cooperativity in Src kinase.

    PubMed

    Foda, Zachariah H; Shan, Yibing; Kim, Eric T; Shaw, David E; Seeliger, Markus A

    2015-01-20

    Protein tyrosine kinases are attractive drug targets because many human diseases are associated with the deregulation of kinase activity. However, how the catalytic kinase domain integrates different signals and switches from an active to an inactive conformation remains incompletely understood. Here we identify an allosteric network of dynamically coupled amino acids in Src kinase that connects regulatory sites to the ATP- and substrate-binding sites. Surprisingly, reactants (ATP and peptide substrates) bind with negative cooperativity to Src kinase while products (ADP and phosphopeptide) bind with positive cooperativity. We confirm the molecular details of the signal relay through the allosteric network by biochemical studies. Experiments on two additional protein tyrosine kinases indicate that the allosteric network may be largely conserved among these enzymes. Our work provides new insights into the regulation of protein tyrosine kinases and establishes a potential conduit by which resistance mutations to ATP-competitive kinase inhibitors can affect their activity.

  20. Transmission and reception of Quad-Carrier QPSK-OFDM signal with blind equalization and overhead-free operation.

    PubMed

    Li, Fan; Zhang, Junwen; Cao, Zizheng; Yu, Jianjun; Li, Xinying; Chen, Lin; Xia, Yan; Chen, Yufei

    2013-12-16

    Quad-Carrier Quadrature Phase Shift Keyed orthogonal frequency division multiplexing (QPSK-OFDM) signal transmission and reception is successfully demonstrated with blind equalization like a 25-ary quadrature amplitude modulation (25-QAM) signal with cascaded multi-modulus algorithm (CMMA) equalization. The phase recovery can be realized with simple Viterbi algorithm and the frequency offset estimation (FOE) should be done with 25-QAM signal before 4-point fast Fourier transform (FFT). 48-Gbit/s Quad-Carrier QPSK-OFDM signal is successfully transmitted over 80-km SMF-28 without penalty.

  1. Prototype positron emission tomography insert with electro-optical signal transmission for simultaneous operation with MRI

    NASA Astrophysics Data System (ADS)

    Olcott, Peter; Kim, Ealgoo; Hong, Keyjo; Lee, Brian J.; Grant, Alexander M.; Chang, Chen-Ming; Glover, Gary; Levin, Craig S.

    2015-05-01

    The simultaneous acquisition of PET and MRI data shows promise to provide powerful capabilities to study disease processes in human subjects, guide the development of novel treatments, and monitor therapy response and disease progression. A brain-size PET detector ring insert for an MRI system is being developed that, if successful, can be inserted into any existing MRI system to enable simultaneous PET and MRI images of the brain to be acquired without mutual interference. The PET insert uses electro-optical coupling to relay all the signals from the PET detectors out of the MRI system using analog modulated lasers coupled to fiber optics. Because the fibers use light instead of electrical signals, the PET detector can be electrically decoupled from the MRI making it partially transmissive to the RF field of the MRI. The SiPM devices and low power lasers were powered using non-magnetic MRI compatible batteries. Also, the number of laser-fiber channels in the system was reduced using techniques adapted from the field of compressed sensing. Using the fact that incoming PET data is sparse in time and space, electronic circuits implementing constant weight codes uniquely encode the detector signals in order to reduce the number of electro-optical readout channels by 8-fold. Two out of a total of sixteen electro-optical detector modules have been built and tested with the entire RF-shielded detector gantry for the PET ring insert. The two detectors have been tested outside and inside of a 3T MRI system to study mutual interference effects and simultaneous performance with MRI. Preliminary results show that the PET insert is feasible for high resolution simultaneous PET/MRI imaging for applications in the brain.

  2. Monitoring allostery in D2O: a necessary control in studies using hydrogen/deuterium exchange to characterize allosteric regulation.

    PubMed

    Prasannan, Charulata B; Artigues, Antonio; Fenton, Aron W

    2011-08-01

    There is currently a renewed focus aimed at understanding allosteric mechanisms at atomic resolution. This current interest seeks to understand how both changes in protein conformations and changes in protein dynamics contribute to relaying an allosteric signal between two ligand binding sites on a protein (e.g., active and allosteric sites). Both nuclear magnetic resonance (NMR), by monitoring protein dynamics directly, and hydrogen/deuterium exchange, by monitoring solvent accessibility of backbone amides, offer insights into protein dynamics. Unfortunately, many allosteric proteins exceed the size limitations of standard NMR techniques. Although hydrogen/deuterium exchange as detected by mass spectrometry (H/DX-MS) offers an alternative evaluation method, any application of hydrogen/deuterium exchange requires that the property being measured functions in both H(2)O and D(2)O. Due to the promising future H/DX-MS has in the evaluation of allosteric mechanisms in large proteins, we demonstrate an evaluation of allosteric regulation in D(2)O. Exemplified using phenylalanine inhibition of rabbit muscle pyruvate kinase, we find that binding of the inhibitor is greatly reduced in D(2)O, but the effector continues to elicit an allosteric response.

  3. An allosteric model for ribonuclease.

    PubMed Central

    Walker, E J; Ralston, G B; Darvey, I G

    1975-01-01

    Data from two assay systems show that the kinetics of the hydrolysis of cytidine 2':3'-cyclic monophosphate by bovine pancreatic RNAase (ribonuclease) is not consistent with conventional models. An allosteric model involving a substrate-dependent change in the equilibrium between two enzyme conformations is proposed. Such a model gives rise to a calculated curve of velocity versus substrate concentration which fits the experimental data. The model is also consistent with the results of an examination of the tryptic digestion of RNAase. Substrate analogues are able to protect RNAase against hydrolysis by trypsin and the percentage of RNAase activity which remains after digestion increases sigmoidally as the analogue concentration is increased. The model also explains the pattern seen in the Km values quoted in the literature and is consistent with strong physical evidence for a ligand-induced conformational change for RNAase reported in the literature. PMID:1167152

  4. Self-powered wireless carbohydrate/oxygen sensitive biodevice based on radio signal transmission.

    PubMed

    Falk, Magnus; Alcalde, Miguel; Bartlett, Philip N; De Lacey, Antonio L; Gorton, Lo; Gutierrez-Sanchez, Cristina; Haddad, Raoudha; Kilburn, Jeremy; Leech, Dónal; Ludwig, Roland; Magner, Edmond; Mate, Diana M; Conghaile, Peter Ó; Ortiz, Roberto; Pita, Marcos; Pöller, Sascha; Ruzgas, Tautgirdas; Salaj-Kosla, Urszula; Schuhmann, Wolfgang; Sebelius, Fredrik; Shao, Minling; Stoica, Leonard; Sygmund, Cristoph; Tilly, Jonas; Toscano, Miguel D; Vivekananthan, Jeevanthi; Wright, Emma; Shleev, Sergey

    2014-01-01

    Here for the first time, we detail self-contained (wireless and self-powered) biodevices with wireless signal transmission. Specifically, we demonstrate the operation of self-sustained carbohydrate and oxygen sensitive biodevices, consisting of a wireless electronic unit, radio transmitter and separate sensing bioelectrodes, supplied with electrical energy from a combined multi-enzyme fuel cell generating sufficient current at required voltage to power the electronics. A carbohydrate/oxygen enzymatic fuel cell was assembled by comparing the performance of a range of different bioelectrodes followed by selection of the most suitable, stable combination. Carbohydrates (viz. lactose for the demonstration) and oxygen were also chosen as bioanalytes, being important biomarkers, to demonstrate the operation of the self-contained biosensing device, employing enzyme-modified bioelectrodes to enable the actual sensing. A wireless electronic unit, consisting of a micropotentiostat, an energy harvesting module (voltage amplifier together with a capacitor), and a radio microchip, were designed to enable the biofuel cell to be used as a power supply for managing the sensing devices and for wireless data transmission. The electronic system used required current and voltages greater than 44 µA and 0.57 V, respectively to operate; which the biofuel cell was capable of providing, when placed in a carbohydrate and oxygen containing buffer. In addition, a USB based receiver and computer software were employed for proof-of concept tests of the developed biodevices. Operation of bench-top prototypes was demonstrated in buffers containing different concentrations of the analytes, showcasing that the variation in response of both carbohydrate and oxygen biosensors could be monitored wirelessly in real-time as analyte concentrations in buffers were changed, using only an enzymatic fuel cell as a power supply.

  5. Self-Powered Wireless Carbohydrate/Oxygen Sensitive Biodevice Based on Radio Signal Transmission

    PubMed Central

    Falk, Magnus; Alcalde, Miguel; Bartlett, Philip N.; De Lacey, Antonio L.; Gorton, Lo; Gutierrez-Sanchez, Cristina; Haddad, Raoudha; Kilburn, Jeremy; Leech, Dónal; Ludwig, Roland; Magner, Edmond; Mate, Diana M.; Conghaile, Peter Ó.; Ortiz, Roberto; Pita, Marcos; Pöller, Sascha; Ruzgas, Tautgirdas; Salaj-Kosla, Urszula; Schuhmann, Wolfgang; Sebelius, Fredrik; Shao, Minling; Stoica, Leonard; Sygmund, Cristoph; Tilly, Jonas; Toscano, Miguel D.; Vivekananthan, Jeevanthi; Wright, Emma; Shleev, Sergey

    2014-01-01

    Here for the first time, we detail self-contained (wireless and self-powered) biodevices with wireless signal transmission. Specifically, we demonstrate the operation of self-sustained carbohydrate and oxygen sensitive biodevices, consisting of a wireless electronic unit, radio transmitter and separate sensing bioelectrodes, supplied with electrical energy from a combined multi-enzyme fuel cell generating sufficient current at required voltage to power the electronics. A carbohydrate/oxygen enzymatic fuel cell was assembled by comparing the performance of a range of different bioelectrodes followed by selection of the most suitable, stable combination. Carbohydrates (viz. lactose for the demonstration) and oxygen were also chosen as bioanalytes, being important biomarkers, to demonstrate the operation of the self-contained biosensing device, employing enzyme-modified bioelectrodes to enable the actual sensing. A wireless electronic unit, consisting of a micropotentiostat, an energy harvesting module (voltage amplifier together with a capacitor), and a radio microchip, were designed to enable the biofuel cell to be used as a power supply for managing the sensing devices and for wireless data transmission. The electronic system used required current and voltages greater than 44 µA and 0.57 V, respectively to operate; which the biofuel cell was capable of providing, when placed in a carbohydrate and oxygen containing buffer. In addition, a USB based receiver and computer software were employed for proof-of concept tests of the developed biodevices. Operation of bench-top prototypes was demonstrated in buffers containing different concentrations of the analytes, showcasing that the variation in response of both carbohydrate and oxygen biosensors could be monitored wirelessly in real-time as analyte concentrations in buffers were changed, using only an enzymatic fuel cell as a power supply. PMID:25310190

  6. Supramolecular Allosteric Cofacial Porphyrin Complexes

    SciTech Connect

    Oliveri, Christopher G.; Gianneschi, Nathan C.; Nguyen, Son Binh T.; Mirkin, Chad A.; Stern, Charlotte L.; Wawrzak, Zdzislaw; Pink, Maren

    2008-04-12

    Nature routinely uses cooperative interactions to regulate cellular activity. For years, chemists have designed synthetic systems that aim toward harnessing the reactivity common to natural biological systems. By learning how to control these interactions in situ, one begins to allow for the preparation of man-made biomimetic systems that can efficiently mimic the interactions found in Nature. To this end, we have designed a synthetic protocol for the preparation of flexible metal-directed supramolecular cofacial porphyrin complexes which are readily obtained in greater than 90% yield through the use of new hemilabile porphyrin ligands with bifunctional ether-phosphine or thioether-phosphine substituents at the 5 and 15 positions on the porphyrin ring. The resulting architectures contain two hemilabile ligand-metal domains (Rh{sup I} or Cu{sup I} sites) and two cofacially aligned porphyrins (Zn{sup II} sites), offering orthogonal functionalities and allowing these multimetallic complexes to exist in two states, 'condensed' or 'open'. Combining the ether-phosphine ligand with the appropriate Rh{sup I} or Cu{sup I} transition-metal precursors results in 'open' macrocyclic products. In contrast, reacting the thioether-phosphine ligand with RhI or CuI precursors yields condensed structures that can be converted into their 'open' macrocyclic forms via introduction of additional ancillary ligands. The change in cavity size that occurs allows these structures to function as allosteric catalysts for the acyl transfer reaction between X-pyridylcarbinol (where X = 2, 3, or 4) and 1-acetylimidazole. For 3- and 4-pyridylcarbinol, the 'open' macrocycle accelerates the acyl transfer reaction more than the condensed analogue and significantly more than the porphyrin monomer. In contrast, an allosteric effect was not observed for 2-pyridylcarbinol, which is expected to be a weaker binder and is unfavorably constrained inside the macrocyclic cavity.

  7. In-Containment Signal Acquisition and Data Transmission via Power Lines within High Dose Areas of Nuclear Power Plants

    SciTech Connect

    Mueller Steffen; Wibbing Sascha; Weigel Robert; Koelpin Alexander; Dennerlein, Juergen; Janke, Iryna; Weber, Johannes

    2015-07-01

    Signal acquisition and data transmission for innovative sensor systems and networks inside the containment of nuclear power plants (NPPs) is still a challenge with respect to safety, performance, reliability, availability, and costs. This especially applies to equipment upgrades for existing plants, special measurements, but also for new builds. This paper presents a novel method for efficient and cost-effective sensor signal acquisition and data transmission via power lines, in order to cope with the disadvantages of common system architectures that often suffer from poor signal integrity due to raw data transmissions via long cables, huge efforts and costs for installation, and low flexibility with respect to maintenance and upgrades. A transmitter-receiver architecture is proposed that allows multiplexing of multiple sensor inputs for unidirectional point-to-point transmission by superimposing information signals on existing AC or DC supply lines, but also on active and inactive sensor wires, or spare cables, using power line communication (PLC) technology. Based on commercial off-the-shelf (COTS) electronic parts, a radiation hard transmitter hardware is designed to operate in harsh environment within the containment during full plant operation. The system's modular approach allows application specific trade-offs between redundancy and throughput regarding data transmission, as well as various sensor input front-ends which are compatible with state of the art systems. PLC technology eliminates the need for costly installation of additional cables and wall penetrations, while providing a complementary and diverse communication technology for upgrades of existing systems. At the receiver side in low dose areas, signals are extracted from the power line, demodulated, and de-multiplexed, in order to regain the original sensor signal information and provide it either in analog or digital output format. Successful laboratory qualification tests, field trails, and a

  8. Shielded balanced and coaxial transmission lines. Parametric measurements and instrumentation relevant to signal waveform transmission in digital service

    NASA Astrophysics Data System (ADS)

    Gans, W. L.; Nahman, N. S.

    1981-06-01

    The impulse and step responses of a shielded cable using time domain terminal measurements and a physically based mathematical model for the transmission line properties of the cable are determined. A computer controlled time domain measurement system was implemented using the automatic pulse measurement system. Data are also developed for the frequency domain complex propagation function (attenuation and its related minimum phase shift). The method is applied to 12 shielded paired conductor (balanced) cables and 5 coaxial cables. Time domain responses are presented for three nominal cable lengths, 60 m (200 ft), 150 m (500 ft), and 300 m (1000 ft).

  9. Influence of signal constellation on the performance of 16-ary DEQAM transmission through a regenerative satellite link

    NASA Astrophysics Data System (ADS)

    Cheung, S. W.

    1990-04-01

    The paper presents the results of a series of computer-simulation tests to determine the effects of nonlinear distortion and adjacent channel interference (ACI), on the tolerance to additive white Gaussian noise (AWGN) of a digital satellite modem. The modem transmits a 16-ary differentially encoded quadrature-amplitude-modulated (16-ary DEQAM) signal over a regenerative satellite link, where the high-power amplifier (HPA) at the transmitter may introduce AM-AM and AM-PM conversion effects into the 16-ary DEQAM signal. Three signal constellations are used at the transmitter, namely (1) a conventional 16-ary DEQAM signal constellation, (2) a predistorted 16-ary DEQAM signal constellation, and (3) a prerotated 16-ary DEQAM signal constellation. An equivalent baseband model of the transmission system is used to determine the performance of the modem under the various conditions studied, and the results are used to select the preferred modem design.

  10. Transmission of blue (S) cone signals through the primate lateral geniculate nucleus

    PubMed Central

    Tailby, C; Szmajda, B A; Buzás, P; Lee, B B; Martin, P R

    2008-01-01

    This study concerns the transmission of short-wavelength-sensitive (S) cone signals through the primate dorsal lateral geniculate nucleus. The principal cell classes, magnocellular (MC) and parvocellular (PC), are traditionally segregated into on- and off-subtypes on the basis of the sign of their response to luminance variation. Cells dominated by input from S-cones (‘blue-on and blue-off’) are less frequently encountered and their properties are less well understood. Here we characterize the spatial and chromatic properties of a large sample of blue-on and blue-off neurons and contrast them with those of PC and MC neurons. The results confirm that blue-on and blue-off cells have larger receptive fields than PC and MC neurons at equivalent eccentricities. Relative to blue-on cells, blue-off cells are less sensitive to S-cone contrast, have larger receptive fields, and show more low-pass spatial frequency tuning. Thus, blue-on and blue-off neurons lack the functional symmetry characteristic of on- and off-subtypes in the MC and PC pathways. The majority of MC and PC cells received no detectible input from S-cones. Where present, input from S-cones tended to provide weak inhibition to PC cells. All cell types showed evidence of a suppressive extra-classical receptive field driven largely or exclusively by ML-cones. These data indicate that S-cone signals are isolated to supply the classical receptive field mechanisms of blue-on and blue-off cells in the LGN, and that the low spatial precision of S-cone vision has origins in both classical and extraclassical receptive field properties of subcortical pathways. PMID:18955378

  11. On the g-protein-coupled receptor heteromers and their allosteric receptor-receptor interactions in the central nervous system: focus on their role in pain modulation.

    PubMed

    Borroto-Escuela, Dasiel O; Romero-Fernandez, Wilber; Rivera, Alicia; Van Craenenbroeck, Kathleen; Tarakanov, Alexander O; Agnati, Luigi F; Fuxe, Kjell

    2013-01-01

    The modulatory role of allosteric receptor-receptor interactions in the pain pathways of the Central Nervous System and the peripheral nociceptors has become of increasing interest. As integrators of nociceptive and antinociceptive wiring and volume transmission signals, with a major role for the opioid receptor heteromers, they likely have an important role in the pain circuits and may be involved in acupuncture. The delta opioid receptor (DOR) exerts an antagonistic allosteric influence on the mu opioid receptor (MOR) function in a MOR-DOR heteromer. This heteromer contributes to morphine-induced tolerance and dependence, since it becomes abundant and develops a reduced G-protein-coupling with reduced signaling mainly operating via β -arrestin2 upon chronic morphine treatment. A DOR antagonist causes a return of the Gi/o binding and coupling to the heteromer and the biological actions of morphine. The gender- and ovarian steroid-dependent recruitment of spinal cord MOR/kappa opioid receptor (KOR) heterodimers enhances antinociceptive functions and if impaired could contribute to chronic pain states in women. MOR1D heterodimerizes with gastrin-releasing peptide receptor (GRPR) in the spinal cord, mediating morphine induced itch. Other mechanism for the antinociceptive actions of acupuncture along meridians may be that it enhances the cross-desensitization of the TRPA1 (chemical nociceptor)-TRPV1 (capsaicin receptor) heteromeric channel complexes within the nociceptor terminals located along these meridians. Selective ionotropic cannabinoids may also produce cross-desensitization of the TRPA1-TRPV1 heteromeric nociceptor channels by being negative allosteric modulators of these channels leading to antinociception and antihyperalgesia.

  12. On the G-Protein-Coupled Receptor Heteromers and Their Allosteric Receptor-Receptor Interactions in the Central Nervous System: Focus on Their Role in Pain Modulation

    PubMed Central

    Borroto-Escuela, Dasiel O.; Romero-Fernandez, Wilber; Rivera, Alicia; Van Craenenbroeck, Kathleen; Tarakanov, Alexander O.; Agnati, Luigi F.; Fuxe, Kjell

    2013-01-01

    The modulatory role of allosteric receptor-receptor interactions in the pain pathways of the Central Nervous System and the peripheral nociceptors has become of increasing interest. As integrators of nociceptive and antinociceptive wiring and volume transmission signals, with a major role for the opioid receptor heteromers, they likely have an important role in the pain circuits and may be involved in acupuncture. The delta opioid receptor (DOR) exerts an antagonistic allosteric influence on the mu opioid receptor (MOR) function in a MOR-DOR heteromer. This heteromer contributes to morphine-induced tolerance and dependence, since it becomes abundant and develops a reduced G-protein-coupling with reduced signaling mainly operating via β-arrestin2 upon chronic morphine treatment. A DOR antagonist causes a return of the Gi/o binding and coupling to the heteromer and the biological actions of morphine. The gender- and ovarian steroid-dependent recruitment of spinal cord MOR/kappa opioid receptor (KOR) heterodimers enhances antinociceptive functions and if impaired could contribute to chronic pain states in women. MOR1D heterodimerizes with gastrin-releasing peptide receptor (GRPR) in the spinal cord, mediating morphine induced itch. Other mechanism for the antinociceptive actions of acupuncture along meridians may be that it enhances the cross-desensitization of the TRPA1 (chemical nociceptor)-TRPV1 (capsaicin receptor) heteromeric channel complexes within the nociceptor terminals located along these meridians. Selective ionotropic cannabinoids may also produce cross-desensitization of the TRPA1-TRPV1 heteromeric nociceptor channels by being negative allosteric modulators of these channels leading to antinociception and antihyperalgesia. PMID:23956775

  13. The influence of the pressure force control signal on selected parameters of the vehicle continuously variable transmission

    NASA Astrophysics Data System (ADS)

    Bieniek, A.; Graba, M.; Prażnowski, K.

    2016-09-01

    The paper presents results of research on the effect of frequency control signal on the course selected operating parameters of the continuously variable transmission CVT. The study used a gear Fuji Hyper M6 with electro-hydraulic control system and proprietary software for control and data acquisition developed in LabView environment.

  14. Regulation of ROS in transmissible gastroenteritis virus-activated apoptotic signaling

    SciTech Connect

    Ding, Li; Zhao, Xiaomin; Huang, Yong; Du, Qian; Dong, Feng; Zhang, Hongling; Song, Xiangjun; Zhang, Wenlong; Tong, Dewen

    2013-12-06

    Highlights: •TGEV infection induced ROS accumulation. •ROS accumulation is involved in TGEV-induced mitochondrial integrity impairment. •ROS is associated with p53 activation and apoptosis occurrence in TGEV-infected cells. -- Abstract: Transmissible gastroenteritis virus (TGEV), an enteropathogenic coronavirus, causes severe lethal watery diarrhea and dehydration in piglets. Previous studies indicate that TGEV infection induces cell apoptosis in host cells. In this study, we investigated the roles and regulation of reactive oxygen species (ROS) in TGEV-activated apoptotic signaling. The results showed that TGEV infection induced ROS accumulation, whereas UV-irradiated TGEV did not promote ROS accumulation. In addition, TGEV infection lowered mitochondrial transmembrane potential in PK-15 cell line, which could be inhibited by ROS scavengers, pyrrolidinedithiocarbamic (PDTC) and N-acetyl-L-cysteine (NAC). Furthermore, the two scavengers significantly inhibited the activation of p38 MAPK and p53 and further blocked apoptosis occurrence through suppressing the TGEV-induced Bcl-2 reduction, Bax redistribution, cytochrome c release and caspase-3 activation. These results suggest that oxidative stress pathway might be a key element in TGEV-induced apoptosis and TGEV pathogenesis.

  15. Intramolecular signal transmission in a tetrameric repressor of the IclR family

    PubMed Central

    Fillet, Sandy; Krell, Tino; Morel, Bertrand; Lu, Duo; Zhang, Xiaodong; Ramos, Juan L.

    2011-01-01

    Members of the IclR family control bacterial genes involved in a number of physiological processes. The IclR-family member TtgV crystallizes as a tetramer, with each TtgV monomer consisting of two domains—a DNA binding domain and an effector recognition domain, which are interconnected by an extended α-helix. When bound to DNA, a kink is introduced so that the extended helix is split in two α-helices (helix-4 and -5). Differential scanning calorimetry studies revealed that TtgV unfolds in a single event, suggesting that the two domains unfold cooperatively. When mutations are introduced in helix-5 that disrupt interactions between Arg98 and Glu102, the thermal unfolding of the TtgV domains becomes uncoupled without compromising effector binding. Two of these mutants (TtgVE102R and TtgVE102A) showed impaired release from target DNA, suggesting that these mutations alter signal transmission. By combining various mutants, we found that the mutations in the connecting α-helix exhibited a dominant effect over mutations in DNA binding and effector binding domains. We propose a model in which the loss of cooperativity of unfolding of TtgV reflects perturbed interdomain communication, and that the transition from the continuous to discontinuous helix may mediate interdomain communication necessary for the proper functioning of TtgV. PMID:21876158

  16. Modelling the transmission of a 40-Gbit s{sup -1} NRZ-ADPSK signal in 50-GHz networks

    SciTech Connect

    Gurkin, N V; Kapin, Yu A; Nanii, Oleg E; Novikov, A G; Pavlov, V N; Plaksin, S O; Plotskii, A Yu; Treshchikov, V N

    2013-06-30

    We study numerically the effect of linear and nonlinear distortions on the transmission of a 40-Gbit s{sup -1} NRZ-ADPSK signal. The results of the numerical simulation coincide within the measurement accuracy with the experimental data, which indicates the adequacy of the mathematical model of the communication line, transmitter and receiver. We have found experimentally and numerically that in communication systems based on a standard telecommunication fibre negative dispersion (approximately -160 ps nm{sup -1}) introduced into the signal being transmitted provides an increase in the maximum admissible signal power coupled into a span and in the value of the admissible attenuation of the spans at 2 dB. It is established that nonlinear distortions caused by the presence of adjacent channels in a frequency network with a step of 50 and 100 GHz (crosstalks) are insignificant in comparison with the interchannel nonlinear distortions. (optical information transmission)

  17. Scalable rule-based modelling of allosteric proteins and biochemical networks.

    PubMed

    Ollivier, Julien F; Shahrezaei, Vahid; Swain, Peter S

    2010-11-04

    Much of the complexity of biochemical networks comes from the information-processing abilities of allosteric proteins, be they receptors, ion-channels, signalling molecules or transcription factors. An allosteric protein can be uniquely regulated by each combination of input molecules that it binds. This "regulatory complexity" causes a combinatorial increase in the number of parameters required to fit experimental data as the number of protein interactions increases. It therefore challenges the creation, updating, and re-use of biochemical models. Here, we propose a rule-based modelling framework that exploits the intrinsic modularity of protein structure to address regulatory complexity. Rather than treating proteins as "black boxes", we model their hierarchical structure and, as conformational changes, internal dynamics. By modelling the regulation of allosteric proteins through these conformational changes, we often decrease the number of parameters required to fit data, and so reduce over-fitting and improve the predictive power of a model. Our method is thermodynamically grounded, imposes detailed balance, and also includes molecular cross-talk and the background activity of enzymes. We use our Allosteric Network Compiler to examine how allostery can facilitate macromolecular assembly and how competitive ligands can change the observed cooperativity of an allosteric protein. We also develop a parsimonious model of G protein-coupled receptors that explains functional selectivity and can predict the rank order of potency of agonists acting through a receptor. Our methodology should provide a basis for scalable, modular and executable modelling of biochemical networks in systems and synthetic biology.

  18. Proposed Mode of Binding and Action of Positive Allosteric Modulators at Opioid Receptors

    PubMed Central

    2016-01-01

    Available crystal structures of opioid receptors provide a high-resolution picture of ligand binding at the primary (“orthosteric”) site, that is, the site targeted by endogenous ligands. Recently, positive allosteric modulators of opioid receptors have also been discovered, but their modes of binding and action remain unknown. Here, we use a metadynamics-based strategy to efficiently sample the binding process of a recently discovered positive allosteric modulator of the δ-opioid receptor, BMS-986187, in the presence of the orthosteric agonist SNC-80, and with the receptor embedded in an explicit lipid–water environment. The dynamics of BMS-986187 were enhanced by biasing the potential acting on the ligand–receptor distance and ligand–receptor interaction contacts. Representative lowest-energy structures from the reconstructed free-energy landscape revealed two alternative ligand binding poses at an allosteric site delineated by transmembrane (TM) helices TM1, TM2, and TM7, with some participation of TM6. Mutations of amino acid residues at these proposed allosteric sites were found to either affect the binding of BMS-986187 or its ability to modulate the affinity and/or efficacy of SNC-80. Taken together, these combined experimental and computational studies provide the first atomic-level insight into the modulation of opioid receptor binding and signaling by allosteric modulators. PMID:26841170

  19. The concept of allosteric interaction and its consequences for the chemistry of the brain.

    PubMed

    Changeux, Jean-Pierre

    2013-09-20

    Throughout this Reflections article, I have tried to follow up on the genesis in the 1960s and subsequent evolution of the concept of allosteric interaction and to examine its consequences within the past decades, essentially in the field of the neuroscience. The main conclusion is that allosteric mechanisms built on similar structural principles operate in bacterial regulatory enzymes, gene repressors (and the related nuclear receptors), rhodopsin, G-protein-coupled receptors, neurotransmitter receptors, ion channels, and so on from prokaryotes up to the human brain yet with important features of their own. Thus, future research on these basic cybernetic sensors is expected to develop in two major directions: at the elementary level, toward the atomic structure and molecular dynamics of the conformational changes involved in signal recognition and transduction, but also at a higher level of organization, the contribution of allosteric mechanisms to the modulation of brain functions.

  20. The Concept of Allosteric Interaction and Its Consequences for the Chemistry of the Brain

    PubMed Central

    Changeux, Jean-Pierre

    2013-01-01

    Throughout this Reflections article, I have tried to follow up on the genesis in the 1960s and subsequent evolution of the concept of allosteric interaction and to examine its consequences within the past decades, essentially in the field of the neuroscience. The main conclusion is that allosteric mechanisms built on similar structural principles operate in bacterial regulatory enzymes, gene repressors (and the related nuclear receptors), rhodopsin, G-protein-coupled receptors, neurotransmitter receptors, ion channels, and so on from prokaryotes up to the human brain yet with important features of their own. Thus, future research on these basic cybernetic sensors is expected to develop in two major directions: at the elementary level, toward the atomic structure and molecular dynamics of the conformational changes involved in signal recognition and transduction, but also at a higher level of organization, the contribution of allosteric mechanisms to the modulation of brain functions. PMID:23878193

  1. Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells

    NASA Astrophysics Data System (ADS)

    Kiyonaka, Shigeki; Kubota, Ryou; Michibata, Yukiko; Sakakura, Masayoshi; Takahashi, Hideo; Numata, Tomohiro; Inoue, Ryuji; Yuzaki, Michisuke; Hamachi, Itaru

    2016-10-01

    The controlled activation of proteins in living cells is an important goal in protein-design research, but to introduce an artificial activation switch into membrane proteins through rational design is a significant challenge because of the structural and functional complexity of such proteins. Here we report the allosteric activation of two types of membrane-bound neurotransmitter receptors, the ion-channel type and the G-protein-coupled glutamate receptors, using coordination chemistry in living cells. The high programmability of coordination chemistry enabled two His mutations, which act as an artificial allosteric site, to be semirationally incorporated in the vicinity of the ligand-binding pockets. Binding of Pd(2,2‧-bipyridine) at the allosteric site enabled the active conformations of the glutamate receptors to be stabilized. Using this approach, we were able to activate selectively a mutant glutamate receptor in live neurons, which initiated a subsequent signal-transduction pathway.

  2. Wavelength reused bidirectional transmission of adaptively modulated optical OFDM signals in WDM-PONs incorporating SOA and RSOA intensity modulators.

    PubMed

    Wei, J L; Hugues-Salas, E; Giddings, R P; Jin, X Q; Zheng, X; Mansoor, S; Tang, J M

    2010-05-10

    Detailed numerical investigations are undertaken of wavelength reused bidirectional transmission of adaptively modulated optical OFDM (AMOOFDM) signals over a single SMF in a colorless WDM-PON incorporating a semiconductor optical amplifier (SOA) intensity modulator and a reflective SOA (RSOA) intensity modulator in the optical line termination and optical network unit, respectively. A comprehensive theoretical model describing the performance of such network scenarios is, for the first time, developed, taking into account dynamic optical characteristics of SOA and RSOA intensity modulators as well as the effects of Rayleigh backscattering (RB) and residual downstream signal-induced crosstalk. The developed model is rigorously verified experimentally in RSOA-based real-time end-to-end OOFDM systems at 7.5 Gb/s. It is shown that the RB noise and crosstalk effects are dominant factors limiting the maximum achievable downstream and upstream transmission performance. Under optimum SOA and RSOA operating conditions as well as practical downstream and upstream optical launch powers, 10 Gb/s downstream and 6 Gb/s upstream over 40 km SMF transmissions of conventional double sideband AMOOFDM signals are feasible without utilizing in-line optical amplification and chromatic dispersion compensation. In particular, the aforementioned transmission performance can be improved to 23 Gb/s downstream and 8 Gb/s upstream over 40 km SMFs when single sideband subcarrier modulation is adopted in the downstream systems.

  3. Demonstration of 2.97-Gb/s video signal transmissions in DML-based IM-DDO-OFDM systems

    NASA Astrophysics Data System (ADS)

    Chen, Ming; He, Jing; Deng, Rui; Chen, Qinghui; Zhang, Jinlong; Chen, Lin

    2016-05-01

    To further investigate the feasibility of the digital signal processing (DSP) algorithms (e.g., symbol timing synchronization, channel estimation and equalization, and sampling clock frequency offset (SCFO) estimation and compensation) for real-time optical orthogonal frequency-division multiplexing (OFDM) system, 2.97-Gb/s real-time high-definition video signal parallel transmission is experimentally demonstrated in OFDM-based short-reach intensity-modulated direct-detection (IM-DD) systems. The experimental results show that, in the presence of ∼12 ppm SCFO between transmitter and receiver, the adaptively modulated OFDM signal transmission over 20 km standard single-mode fiber with an error bit rate less than 1 × 10-9 can be achieved by using only DSP-based small SCFO estimation and compensation method without utilizing forward error correction technique. To the best of our knowledge, for the first time, we successfully demonstrate that the video signal at a bit rate in excess of 1-Gb/s transmission in a simple real-valued inverse fast Fourier transform and fast Fourier transform based IM-DD optical OFDM system employing a directly modulated laser.

  4. Signal transmission in a human body medium-based body sensor network using a Mach-Zehnder electro-optical sensor.

    PubMed

    Song, Yong; Hao, Qun; Zhang, Kai; Wang, Jingwen; Jin, Xuefeng; Sun, He

    2012-11-30

    The signal transmission technology based on the human body medium offers significant advantages in Body Sensor Networks (BSNs) used for healthcare and the other related fields. In previous works we have proposed a novel signal transmission method based on the human body medium using a Mach-Zehnder electro-optical (EO) sensor. In this paper, we present a signal transmission system based on the proposed method, which consists of a transmitter, a Mach-Zehnder EO sensor and a corresponding receiving circuit. Meanwhile, in order to verify the frequency response properties and determine the suitable parameters of the developed system, in-vivo measurements have been implemented under conditions of different carrier frequencies, baseband frequencies and signal transmission paths. Results indicate that the proposed system will help to achieve reliable and high speed signal transmission of BSN based on the human body medium.

  5. 40-Gb/s PDM-QPSK signal transmission over 160-m wireless distance at W-band.

    PubMed

    Xiao, Jiangnan; Yu, Jianjun; Li, Xinying; Xu, Yuming; Zhang, Ziran; Chen, Long

    2015-03-15

    We experimentally demonstrate a W-band optical-wireless transmission system over 160-m wireless distance with a bit rate up to 40 Gb/s. The optical-wireless transmission system adopts optical polarization-division-multiplexing (PDM), multiple-input multiple-output (MIMO) reception and antenna polarization diversity. Using this system, we experimentally demonstrate the 2×2 MIMO wireless delivery of 20- and 40-Gb/s PDM quadrature-phase-shift-keying (PDM-QPSK) signals over 640- and 160-m wireless links, respectively. The bit-error ratios (BERs) of these transmission systems are both less than the forward-error-correction (FEC) threshold of 3.8×10-3.

  6. Demostration of 520 Gb/s/λ pre-equalized DFT-spread PDM-16QAM-OFDM signal transmission.

    PubMed

    Li, Fan; Yu, Jianjun; Cao, Zizheng; Chen, Ming; Zhang, Junwen; Li, Xinying

    2016-02-08

    In this paper, we successfully transmit 8 × 520 Gb/s pre-equalized DFT-spread PDM-16QAM orthogonal frequency-division multiplexing (OFDM) signal over 840 km SMF with BER under 2.4 × 10(-2). We discuss how to obtain accurate tranceivers' response during pre-equalization for DFT-spread OFDM with coherent detection and we find conventional OFDM symbols training sequences (TSs) outperform DFT-spread OFDM symbols TSs in obtaining channel response for pre-equalization and equalization. Additionally, the optimal IFFT/FFT size is explored for the pre-equalized DFT-spread PDM-16QAM-OFDM transmission systems. It is the first time to realize 400 Gb/s/λ net rate OFDM signal transmission.

  7. Prepaying the entropic cost for allosteric regulation in KIX.

    PubMed

    Law, Sean M; Gagnon, Jessica K; Mapp, Anna K; Brooks, Charles L

    2014-08-19

    The kinase-inducible domain interacting (KIX) domain of the CREB binding protein (CBP) is capable of simultaneously binding two intrinsically disordered transcription factors, such as the mixed-lineage leukemia (MLL) and c-Myb peptides, at isolated interaction sites. In vitro, the affinity for binding c-Myb is approximately doubled when KIX is in complex with MLL, which suggests a positive cooperative binding mechanism, and the affinity for MLL is also slightly increased when KIX is first bound by c-Myb. Expanding the scope of recent NMR and computational studies, we explore the allosteric mechanism at a detailed molecular level that directly connects the microscopic structural dynamics to the macroscopic shift in binding affinities. To this end, we have performed molecular dynamics simulations of free KIX, KIX-c-Myb, MLL-KIX, and MLL-KIX-c-Myb using a topology-based Gō-like model. Our results capture an increase in affinity for the peptide in the allosteric site when KIX is prebound by a complementary effector and both peptides follow an effector-independent folding-and-binding mechanism. More importantly, we discover that MLL binding lowers the entropic cost for c-Myb binding, and vice versa, by stabilizing the L12-G2 loop and the C-terminal region of the α3 helix on KIX. This work demonstrates the importance of entropy in allosteric signaling between promiscuous molecular recognition sites and can inform the rational design of small molecule stabilizers to target important regions of conformationally dynamic proteins.

  8. Designing Allosteric Control into Enzymes by Chemical Rescue of Structure

    SciTech Connect

    Deckert, Katelyn; Budiardjo, S. Jimmy; Brunner, Luke C.; Lovell, Scott; Karanicolas, John

    2012-08-07

    Ligand-dependent activity has been engineered into enzymes for purposes ranging from controlling cell morphology to reprogramming cellular signaling pathways. Where these successes have typically fused a naturally allosteric domain to the enzyme of interest, here we instead demonstrate an approach for designing a de novo allosteric effector site directly into the catalytic domain of an enzyme. This approach is distinct from traditional chemical rescue of enzymes in that it relies on disruption and restoration of structure, rather than active site chemistry, as a means to achieve modulate function. We present two examples, W33G in a {beta}-glycosidase enzyme ({beta}-gly) and W492G in a {beta}-glucuronidase enzyme ({beta}-gluc), in which we engineer indole-dependent activity into enzymes by removing a buried tryptophan side chain that serves as a buttress for the active site architecture. In both cases, we observe a loss of function, and in both cases we find that the subsequent addition of indole can be used to restore activity. Through a detailed analysis of {beta}-gly W33G kinetics, we demonstrate that this rescued enzyme is fully functionally equivalent to the corresponding wild-type enzyme. We then present the apo and indole-bound crystal structures of {beta}-gly W33G, which together establish the structural basis for enzyme inactivation and rescue. Finally, we use this designed switch to modulate {beta}-glycosidase activity in living cells using indole. Disruption and recovery of protein structure may represent a general technique for introducing allosteric control into enzymes, and thus may serve as a starting point for building a variety of bioswitches and sensors.

  9. Probing the Sophisticated Synergistic Allosteric Regulation of Aromatic Amino Acid Biosynthesis in Mycobacterium tuberculosis Using ᴅ-Amino Acids

    PubMed Central

    Reichau, Sebastian; Blackmore, Nicola J.; Jiao, Wanting; Parker, Emily J.

    2016-01-01

    Chirality plays a major role in recognition and interaction of biologically important molecules. The enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAH7PS) is the first enzyme of the shikimate pathway, which is responsible for the synthesis of aromatic amino acids in bacteria and plants, and a potential target for the development of antibiotics and herbicides. DAH7PS from Mycobacterium tuberculosis (MtuDAH7PS) displays an unprecedented complexity of allosteric regulation, with three interdependent allosteric binding sites and a ternary allosteric response to combinations of the aromatic amino acids l-Trp, l-Phe and l-Tyr. In order to further investigate the intricacies of this system and identify key residues in the allosteric network of MtuDAH7PS, we studied the interaction of MtuDAH7PS with aromatic amino acids that bear the non-natural d-configuration, and showed that the d-amino acids do not elicit an allosteric response. We investigated the binding mode of d-amino acids using X-ray crystallography, site directed mutagenesis and isothermal titration calorimetry. Key differences in the binding mode were identified: in the Phe site, a hydrogen bond between the amino group of the allosteric ligands to the side chain of Asn175 is not established due to the inverted configuration of the ligands. In the Trp site, d-Trp forms no interaction with the main chain carbonyl group of Thr240 and less favourable interactions with Asn237 when compared to the l-Trp binding mode. Investigation of the MtuDAH7PSN175A variant further supports the hypothesis that the lack of key interactions in the binding mode of the aromatic d-amino acids are responsible for the absence of an allosteric response, which gives further insight into which residues of MtuDAH7PS play a key role in the transduction of the allosteric signal. PMID:27128682

  10. Characterization of the novel positive allosteric modulator, LY2119620, at the muscarinic M(2) and M(4) receptors.

    PubMed

    Croy, Carrie H; Schober, Douglas A; Xiao, Hongling; Quets, Anne; Christopoulos, Arthur; Felder, Christian C

    2014-07-01

    The M(4) receptor is a compelling therapeutic target, as this receptor modulates neural circuits dysregulated in schizophrenia, and there is clinical evidence that muscarinic agonists possess both antipsychotic and procognitive efficacy. Recent efforts have shifted toward allosteric ligands to maximize receptor selectivity and manipulate endogenous cholinergic and dopaminergic signaling. In this study, we present the pharmacological characterization of LY2119620 (3-amino-5-chloro-N-cyclopropyl-4-methyl-6-[2-(4-methylpiperazin-1-yl)-2-oxoethoxy] thieno[2,3-b]pyridine-2-carboxamide), a M(2)/M(4) receptor-selective positive allosteric modulator (PAM), chemically evolved from hits identified through a M4 allosteric functional screen. Although unsuitable as a therapeutic due to M(2) receptor cross-reactivity and, thus, potential cardiovascular liability, LY2119620 surpassed previous congeners in potency and PAM activity and broadens research capabilities through its development into a radiotracer. Characterization of LY2119620 revealed evidence of probe dependence in both binding and functional assays. Guanosine 5'-[γ-(35)S]-triphosphate assays displayed differential potentiation depending on the orthosteric-allosteric pairing, with the largest cooperativity observed for oxotremorine M (Oxo-M) LY2119620. Further [(3)H]Oxo-M saturation binding, including studies with guanosine-5'-[(β,γ)-imido]triphosphate, suggests that both the orthosteric and allosteric ligands can alter the population of receptors in the active G protein-coupled state. Additionally, this work expands the characterization of the orthosteric agonist, iperoxo, at the M(4) receptor, and demonstrates that an allosteric ligand can positively modulate the binding and functional efficacy of this high efficacy ligand. Ultimately, it was the M(2) receptor pharmacology and PAM activity with iperoxo that made LY2119620 the most suitable allosteric partner for the M(2) active-state structure recently solved

  11. Probing the Sophisticated Synergistic Allosteric Regulation of Aromatic Amino Acid Biosynthesis in Mycobacterium tuberculosis Using ᴅ-Amino Acids.

    PubMed

    Reichau, Sebastian; Blackmore, Nicola J; Jiao, Wanting; Parker, Emily J

    2016-01-01

    Chirality plays a major role in recognition and interaction of biologically important molecules. The enzyme 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DAH7PS) is the first enzyme of the shikimate pathway, which is responsible for the synthesis of aromatic amino acids in bacteria and plants, and a potential target for the development of antibiotics and herbicides. DAH7PS from Mycobacterium tuberculosis (MtuDAH7PS) displays an unprecedented complexity of allosteric regulation, with three interdependent allosteric binding sites and a ternary allosteric response to combinations of the aromatic amino acids l-Trp, l-Phe and l-Tyr. In order to further investigate the intricacies of this system and identify key residues in the allosteric network of MtuDAH7PS, we studied the interaction of MtuDAH7PS with aromatic amino acids that bear the non-natural d-configuration, and showed that the d-amino acids do not elicit an allosteric response. We investigated the binding mode of d-amino acids using X-ray crystallography, site directed mutagenesis and isothermal titration calorimetry. Key differences in the binding mode were identified: in the Phe site, a hydrogen bond between the amino group of the allosteric ligands to the side chain of Asn175 is not established due to the inverted configuration of the ligands. In the Trp site, d-Trp forms no interaction with the main chain carbonyl group of Thr240 and less favourable interactions with Asn237 when compared to the l-Trp binding mode. Investigation of the MtuDAH7PSN175A variant further supports the hypothesis that the lack of key interactions in the binding mode of the aromatic d-amino acids are responsible for the absence of an allosteric response, which gives further insight into which residues of MtuDAH7PS play a key role in the transduction of the allosteric signal.

  12. The Structure of KinA-Sda Complex Suggests An Allosteric Mechanism of Histidine Kinase Inhibition

    SciTech Connect

    Whitten, A.E.; Jacques, D.A.; Hammouda, B.; Hanley, T.; King, G.F.; Guss, J.Mitchell.; Trewhella, J.; Langley, D.B.; /Sydney U. /NIST, Wash., D.C. /Utah U.

    2007-07-13

    The Bacillus subtilis histidine kinase KinA controls activation of the transcription factor governing sporulation, Spo0A. The decision to sporulate involves KinA phosphorylating itself on a conserved histidine residue, after which the phosphate moiety is relayed via two other proteins to Spo0A. The DNA-damage checkpoint inhibitor Sda halts this pathway by binding KinA and blocking the autokinase reaction. We have performed small-angle X-ray scattering and neutron contrast variation studies on the complex formed by KinA and Sda. The data show that two Sda molecules bind to the base of the DHp dimerization domain of the KinA dimer. In this position Sda does not appear to be able to sterically block the catalytic domain from accessing its target histidine, as previously proposed, but rather may effect an allosteric mode of inhibition involving transmission of the inhibitory signal via the four-helix bundle that forms the DHp domain.

  13. Demonstration of DFT-spread 256QAM-OFDM signal transmission with cost-effective directly modulated laser.

    PubMed

    Li, Fan; Yu, Jianjun; Fang, Yuan; Dong, Ze; Li, Xinying; Chen, Lin

    2014-04-07

    We experimentally demonstrated a 256-ary quadrature amplitude modulation (256QAM) direct-detection optical orthogonal frequency division multiplexing (DDO-OFDM) transmission system utilizing a cost-effective directly modulated laser (DML). Intra-symbol frequency-domain averaging (ISFA) is applied to suppress in-band noise while the channel response estimation and Discrete Fourier Transform-spread (DFT-spread) is used to reduce the peak-to-average power ratio (PAPR) of the transmitted OFDM signal. The bit-error ratio (BER) of 15-Gbit/s 256QAM-OFDM signal has been measured after 20-km SSMF transmission that is less than 7% forward-error-correction (FEC) threshold of 3.8 × 10(-3) as the launch power into fiber is set at 6dBm. For 11.85-Gbit/s 256QAM-OFDM signal, with the aid of ISFA-based channel estimation and PAPR reduction enabled by DFT-spread, the BER after 20-km SSMF transmission can be improved from 6.4 × 10(-3) to 6.8 × 10(-4) when the received optical power is -6dBm.

  14. Sensitivity and kinetics of signal transmission at the first visual synapse differentially impact visually-guided behavior

    PubMed Central

    Sarria, Ignacio; Pahlberg, Johan; Cao, Yan; Kolesnikov, Alexander V; Kefalov, Vladimir J; Sampath, Alapakkam P; Martemyanov, Kirill A

    2015-01-01

    In the retina, synaptic transmission between photoreceptors and downstream ON-bipolar neurons (ON-BCs) is mediated by a GPCR pathway, which plays an essential role in vision. However, the mechanisms that control signal transmission at this synapse and its relevance to behavior remain poorly understood. In this study we used a genetic system to titrate the rate of GPCR signaling in ON-BC dendrites by varying the concentration of key RGS proteins and measuring the impact on transmission of signal between photoreceptors and ON-BC neurons using electroretinography and single cell recordings. We found that sensitivity, onset timing, and the maximal amplitude of light-evoked responses in rod- and cone-driven ON-BCs are determined by different RGS concentrations. We further show that changes in RGS concentration differentially impact visually guided-behavior mediated by rod and cone ON pathways. These findings illustrate that neuronal circuit properties can be modulated by adjusting parameters of GPCR-based neurotransmission at individual synapses. DOI: http://dx.doi.org/10.7554/eLife.06358.001 PMID:25879270

  15. Boronic acids as probes for investigation of allosteric modulation of the chemokine receptor CXCR3.

    PubMed

    Bernat, Viachaslau; Admas, Tizita Haimanot; Brox, Regine; Heinemann, Frank W; Tschammer, Nuska

    2014-11-21

    The chemokine receptor CXCR3 is a G protein-coupled receptor, which conveys extracellular signals into cells by changing its conformation upon agonist binding. To facilitate the mechanistic understanding of allosteric modulation of CXCR3, we combined computational modeling with the synthesis of novel chemical tools containing boronic acid moiety, site-directed mutagenesis, and detailed functional characterization. The design of boronic acid derivatives was based on the predictions from homology modeling and docking. The choice of the boronic acid moiety was dictated by its unique ability to interact with proteins in a reversible covalent way, thereby influencing conformational dynamics of target biomolecules. During the synthesis of the library we have developed a novel approach for the purification of drug-like boronic acids. To validate the predicted binding mode and to identify amino acid residues responsible for the transduction of signal through CXCR3, we conducted a site-directed mutagenesis study. With the use of allosteric radioligand RAMX3 we were able to establish the existence of a second allosteric binding pocket in CXCR3, which enables different binding modes of structurally closely related allosteric modulators of CXCR3. We have also identified residues Trp109(2.60) and Lys300(7.35) inside the transmembrane bundle of the receptor as crucial for the regulation of the G protein activation. Furthermore, we report the boronic acid 14 as the first biased negative allosteric modulator of the receptor. Overall, our data demonstrate that boronic acid derivatives represent an outstanding tool for determination of key receptor-ligand interactions and induction of ligand-biased signaling.

  16. Flower development in the passion fruit Passiflora edulis requires a photoperiod-induced systemic graft-transmissible signal.

    PubMed

    Nave, Nahum; Katz, Ehud; Chayut, Noam; Gazit, Shmuel; Samach, Alon

    2010-12-01

    Different organisms use gradual seasonal changes in photoperiod to correctly time diverse developmental processes, such as transition to flowering in plants. Florigen is a systemic signal formed in leaves exposed to specific environmental cues, mainly photoperiodic, and capable of triggering flower induction in several species. Here we show that in Passiflora edulis, a perennial climbing vine, flower initiation occurs throughout the year; however, without long photoperiods, flower primordia show arrested growth and differentiation at an early stage. Our results support the existence of a positive, systemic, graft-transmissible signal, produced in mature leaves under LDs, that is required for normal flower development beyond sepal formation. Our results also suggest that Gibberellin acts to inhibit flower development. We provide evidence for genetic variation in the response to short photoperiods. A genotype capable of forming developed flowers under short photoperiods produces a positive graft transmissible signal allowing normal flower development under short days in a cultivar which normally aborts flower development under these conditions. We believe these findings contribute towards discovering the chemical nature of this interesting mobile signal involved in flower development.

  17. Seamless integration of 57.2-Gb/s signal wireline transmission and 100-GHz wireless delivery.

    PubMed

    Li, Xinying; Yu, Jianjun; Dong, Ze; Cao, Zizheng; Chi, Nan; Zhang, Junwen; Shao, Yufeng; Tao, Li

    2012-10-22

    We experimentally demonstrated the seamless integration of 57.2-Gb/s signal wireline transmission and 100-GHz wireless delivery adopting polarization-division-multiplexing quadrature-phase-shift-keying (PDM-QPSK) modulation with 400-km single-mode fiber-28 (SMF-28) transmission and 1-m wireless delivery. The X- and Y-polarization components of optical PDM-QPSK baseband signal are simultaneously up-converted to 100 GHz by optical polarization-diversity heterodyne beating, and then independently transmitted and received by two pairs of transmitter and receiver antennas, which make up a 2x2 multiple-input multiple-output (MIMO) wireless link based on microwave polarization multiplexing. At the wireless receiver, a two-stage down conversion is firstly done in analog domain based on balanced mixer and sinusoidal radio frequency (RF) signal, and then in digital domain based on digital signal processing (DSP). Polarization de-multiplexing is realized by constant modulus algorithm (CMA) based on DSP in heterodyne coherent detection. Our experimental results show that more taps are required for CMA when the X- and Y-polarization antennas have different wireless distance.

  18. Cell speed, persistence and information transmission during signal relay and collective migration.

    PubMed

    McCann, Colin P; Kriebel, Paul W; Parent, Carole A; Losert, Wolfgang

    2010-05-15

    Collective migration is a key feature of the social amoebae Dictyostelium discoideum, where the binding of chemoattractants leads to the production and secretion of additional chemoattractant and the relay of the signal to neighboring cells. This then guides cells to migrate collectively in a head-to-tail fashion. We used mutants that were defective in signal relay to elucidate which quantitative metrics of cell migration are most strongly affected by signal relay and collective motion. We show that neither signal relay nor collective motion markedly impact the speed of cell migration. Cells maintained a preferred overall direction of motion for several minutes with similar persistence, regardless of whether or not they were attracted to moving neighbors, moving collectively in contact with their neighbors, or simply following a fixed exogenous signal. We quantitatively establish that signal relay not only increases the number of cells that respond to a chemotactic signal, but most remarkably, also transmits information about the location of the source accurately over large distances, independently of the strength of the exogenous signal. We envision that signal relay has a similar key role in the migration of a variety of chemotaxing mammalian cells that can relay chemoattractant signals.

  19. Allosteric substrate switching in a voltage-sensing lipid phosphatase.

    PubMed

    Grimm, Sasha S; Isacoff, Ehud Y

    2016-04-01

    Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We found that the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), has not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage-sensing domain (VSD). Using fast fluorescence resonance energy transfer (FRET) reporters of PIPs to monitor enzyme activity and voltage-clamp fluorometry to monitor conformational changes in the VSD, we found that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage-sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This two-step allosteric control over a dual-specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility, endocytosis and exocytosis.

  20. Allosteric substrate switching in a voltage sensing lipid phosphatase

    PubMed Central

    Grimm, Sasha S.; Isacoff, Ehud Y.

    2016-01-01

    Allostery provides a critical control over enzyme activity, biasing the catalytic site between inactive and active states. We find the Ciona intestinalis voltage-sensing phosphatase (Ci-VSP), which modifies phosphoinositide signaling lipids (PIPs), to have not one but two sequential active states with distinct substrate specificities, whose occupancy is allosterically controlled by sequential conformations of the voltage sensing domain (VSD). Using fast FRET reporters of PIPs to monitor enzyme activity and voltage clamp fluorometry to monitor conformational changes in the VSD, we find that Ci-VSP switches from inactive to a PIP3-preferring active state when the VSD undergoes an initial voltage sensing motion and then into a second PIP2-preferring active state when the VSD activates fully. This novel 2-step allosteric control over a dual specificity enzyme enables voltage to shape PIP concentrations in time, and provides a mechanism for the complex modulation of PIP-regulated ion channels, transporters, cell motility and endo/exocytosis. PMID:26878552

  1. Photonic-aided pre-coding QAM signal transmission in multi-antenna radio over fiber system

    NASA Astrophysics Data System (ADS)

    Zhang, Qi; Yu, Jianjun; Li, Xinying; Zhu, Ming; Xin, Xiangjun; Chang, Gee-Kung

    2015-11-01

    A novel method has been proposed and experimentally demonstrated to provide photonic-aided pre-coding quadrature amplitude modulation (QAM) signal transmission in multi-antenna radio over fiber system to increase the throughput. For downlink, two different multi-level amplitude-shift-keying (M-ASK) modulated signals, such as 4-ASK signals, are applied on two uncorrelated optical Mach-Zehnder modulators at central office. After photonic-aided pre-coding module and photo-detection process, the received M-ASK mm-waves from two remote access units (RAUs) can be synthesized to a M2-QAM signal in the proposed system. Regardless of forward error correction (FEC) coding overhead, the 4-Gb/s and 8-Gb/s 16-QAM mm-wave signals are obtained from two independent 2-Gb/s and 4-Gb/s 4-ASK 40-GHz channels, respectively. The experimental results show that a doubled bit rate of the original 4-ASK one can be achieved without additional digital signal processing (DSP) in small cell RAUs and mobile users.

  2. Dancing through Life: Molecular Dynamics Simulations and Network-Centric Modeling of Allosteric Mechanisms in Hsp70 and Hsp110 Chaperone Proteins

    PubMed Central

    Stetz, Gabrielle; Verkhivker, Gennady M.

    2015-01-01

    Hsp70 and Hsp110 chaperones play an important role in regulating cellular processes that involve protein folding and stabilization, which are essential for the integrity of signaling networks. Although many aspects of allosteric regulatory mechanisms in Hsp70 and Hsp110 chaperones have been extensively studied and significantly advanced in recent experimental studies, the atomistic picture of signal propagation and energetics of dynamics-based communication still remain unresolved. In this work, we have combined molecular dynamics simulations and protein stability analysis of the chaperone structures with the network modeling of residue interaction networks to characterize molecular determinants of allosteric mechanisms. We have shown that allosteric mechanisms of Hsp70 and Hsp110 chaperones may be primarily determined by nucleotide-induced redistribution of local conformational ensembles in the inter-domain regions and the substrate binding domain. Conformational dynamics and energetics of the peptide substrate binding with the Hsp70 structures has been analyzed using free energy calculations, revealing allosteric hotspots that control negative cooperativity between regulatory sites. The results have indicated that cooperative interactions may promote a population-shift mechanism in Hsp70, in which functional residues are organized in a broad and robust allosteric network that can link the nucleotide-binding site and the substrate-binding regions. A smaller allosteric network in Hsp110 structures may elicit an entropy-driven allostery that occurs in the absence of global structural changes. We have found that global mediating residues with high network centrality may be organized in stable local communities that are indispensable for structural stability and efficient allosteric communications. The network-centric analysis of allosteric interactions has also established that centrality of functional residues could correlate with their sensitivity to mutations

  3. Optical frequency upconversion technique for transmission of wireless MIMO-type signals over optical fiber.

    PubMed

    Shaddad, R Q; Mohammad, A B; Al-Gailani, S A; Al-Hetar, A M

    2014-01-01

    The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF) technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO) signals feeding multiple antennas in the fiber wireless (FiWi) system. A novel optical frequency upconversion (OFU) technique is proposed to solve this problem. In this paper, the novel OFU approach is used to transmit three wireless MIMO signals over a 20 km standard single mode fiber (SMF). The OFU technique exploits one optical source to produce multiple wavelengths by delivering it to a LiNbO3 external optical modulator. The wireless MIMO signals are then modulated by LiNbO3 optical intensity modulators separately using the generated optical carriers from the OFU process. These modulators use the optical single-sideband with carrier (OSSB+C) modulation scheme to optimize the system performance against the fiber dispersion effect. Each wireless MIMO signal is with a 2.4 GHz or 5 GHz carrier frequency, 1 Gb/s data rate, and 16-quadrature amplitude modulation (QAM). The crosstalk between the wireless MIMO signals is highly suppressed, since each wireless MIMO signal is carried on a specific optical wavelength.

  4. 47 CFR 11.51 - EAS code and Attention Signal Transmission requirements.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... section on at least one channel. The Attention signal may be produced from a storage device. Additionally... Attention signal may be produced from a storage device. Additionally, these analog cable systems, digital... deployed an Intermediary Device to meet its CAP-related obligations, this requirement shall be...

  5. 47 CFR 11.51 - EAS code and Attention Signal Transmission requirements.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... section on at least one channel. The Attention signal may be produced from a storage device. Additionally... Attention signal may be produced from a storage device. Additionally, these analog cable systems, digital... deployed an Intermediary Device to meet its CAP-related obligations, this requirement shall be...

  6. Optical Frequency Upconversion Technique for Transmission of Wireless MIMO-Type Signals over Optical Fiber

    PubMed Central

    Shaddad, R. Q.; Mohammad, A. B.; Al-Gailani, S. A.; Al-Hetar, A. M.

    2014-01-01

    The optical fiber is well adapted to pass multiple wireless signals having different carrier frequencies by using radio-over-fiber (ROF) technique. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber, such as wireless multi-input multi-output (MIMO) signals feeding multiple antennas in the fiber wireless (FiWi) system. A novel optical frequency upconversion (OFU) technique is proposed to solve this problem. In this paper, the novel OFU approach is used to transmit three wireless MIMO signals over a 20 km standard single mode fiber (SMF). The OFU technique exploits one optical source to produce multiple wavelengths by delivering it to a LiNbO3 external optical modulator. The wireless MIMO signals are then modulated by LiNbO3 optical intensity modulators separately using the generated optical carriers from the OFU process. These modulators use the optical single-sideband with carrier (OSSB+C) modulation scheme to optimize the system performance against the fiber dispersion effect. Each wireless MIMO signal is with a 2.4 GHz or 5 GHz carrier frequency, 1 Gb/s data rate, and 16-quadrature amplitude modulation (QAM). The crosstalk between the wireless MIMO signals is highly suppressed, since each wireless MIMO signal is carried on a specific optical wavelength. PMID:24772009

  7. Nonlinear impairment compensation for DFT-S OFDM signal transmission with directly modulated laser and direct detection

    NASA Astrophysics Data System (ADS)

    Gou, Pengqi; Wang, Kaihui; Qin, Chaoyi; Yu, Jianjun

    2017-03-01

    We experimentally demonstrate a 16-ary quadrature amplitude modulation (16QAM) DFT-spread optical orthogonal frequency division multiplexing (OFDM) transmission system utilizing a cost-effective directly modulated laser (DML) and direct detection. For 20-Gbaud 16QAM-OFDM signal, with the aid of nonlinear equalization (NLE) algorithm, we respectively provide 6.2-dB and 5.2-dB receiver sensitivity improvement under the hard-decision forward-error-correction (HD-FEC) threshold of 3.8×10-3 for the back-to-back (BTB) case and after transmission over 10-km standard single mode fiber (SSMF) case, related to only adopt post-equalization scheme. To our knowledge, this is the first time to use dynamic nonlinear equalizer (NLE) based on the summation of the square of the difference between samples in one IM/DD OFDM system with DML to mitigate nonlinear distortion.

  8. In-Containment Signal Conditioning and Transmission via Power Lines within High Dose Rate Areas of Nuclear Power Plants

    SciTech Connect

    Mueller, Steffen; Weigel, Robert; Koelpin, Alexander; Dennerlein, Juergen; Janke, Iryna; Weber, Johannes

    2015-07-01

    Signal conditioning and transmission for sensor systems and networks within the containment of nuclear power plants (NPPs) still poses a challenge to engineers, particularly in the case of equipment upgrades for existing plants, temporary measurements, decommissioning of plants, but also for new builds. This paper presents an innovative method for efficient and cost-effective instrumentation within high dose rate areas inside the containment. A transmitter-receiver topology is proposed that allows simultaneous, unidirectional point-to-point transmission of multiple sensor signals by superimposing them on existing AC or DC power supply cables using power line communication (PLC) technology. Thereby the need for costly installation of additional cables and containment penetrations is eliminated. Based on commercial off-the-shelf (COTS) electronic parts, a radiation hard transmitter is designed to operate in harsh environment within the containment during full plant operation. Hardware modularity of the transmitter allows application specific tradeoffs between redundancy and channel bandwidth. At receiver side in non-radiated areas, signals are extracted from the power line, demodulated, and provided either in analog or digital output format. Laboratory qualification tests and field test results within a boiling water reactor (BWR) are validating the proof of concept of the proposed system. (authors)

  9. Contribution of rpfB to cell-to-cell signal synthesis, virulence, and vector transmission of Xylella fastidiosa.

    PubMed

    Almeida, Rodrigo P P; Killiny, Nabil; Newman, Karyn L; Chatterjee, Subhadeep; Ionescu, Michael; Lindow, Steven E

    2012-04-01

    In Xylella fastidiosa the fatty acid signal molecule diffusible signaling factor (DSF) is produced and sensed by components of the regulation of pathogenicity factors (rpf) cluster; lack of DSF production in RpfF mutants results in a non-vector-transmissible phenotype yet cells are hypervirulent to grape. rpfB has not been characterized in Xylella fastidiosa, although its homolog has been suggested to be required for DSF synthesis in Xanthomonas campestris pv. campestris. We show that RpfB is involved in DSF processing in both Xylella fastidiosa and Xanthomonas campestris, affecting the profile of DSF-like fatty acids observed in thin-layer chromatography. Although three fatty acids whose production is dependent on RpfF were detected in Xylella fastidiosa and Xanthomonas campestris wild-type strains, their respective rpfB mutants accumulated primarily one chemical species. Although no quantifiable effect of rpfB on plant colonization by Xylella fastidiosa was found, insect colonization and transmission was reduced. Thus, RpfB apparently is involved in DSF processing, and like Xanthomonas campestris, Xylella fastidiosa also produces multiple DSF molecules. It is possible that Xylella fastidiosa coordinates host vector and plant colonization by varying the proportions of different forms of DSF signals via RpfB.

  10. Allosteric and hyperekplexic mutant phenotypes investigated on an α1 glycine receptor transmembrane structure.

    PubMed

    Moraga-Cid, Gustavo; Sauguet, Ludovic; Huon, Christèle; Malherbe, Laurie; Girard-Blanc, Christine; Petres, Stéphane; Murail, Samuel; Taly, Antoine; Baaden, Marc; Delarue, Marc; Corringer, Pierre-Jean

    2015-03-03

    The glycine receptor (GlyR) is a pentameric ligand-gated ion channel (pLGIC) mediating inhibitory transmission in the nervous system. Its transmembrane domain (TMD) is the target of allosteric modulators such as general anesthetics and ethanol and is a major locus for hyperekplexic congenital mutations altering the allosteric transitions of activation or desensitization. We previously showed that the TMD of the human α1GlyR could be fused to the extracellular domain of GLIC, a bacterial pLGIC, to form a functional chimera called Lily. Here, we overexpress Lily in Schneider 2 insect cells and solve its structure by X-ray crystallography at 3.5 Å resolution. The TMD of the α1GlyR adopts a closed-channel conformation involving a single ring of hydrophobic residues at the center of the pore. Electrophysiological recordings show that the phenotypes of key allosteric mutations of the α1GlyR, scattered all along the pore, are qualitatively preserved in this chimera, including those that confer decreased sensitivity to agonists, constitutive activity, decreased activation kinetics, or increased desensitization kinetics. Combined structural and functional data indicate a pore-opening mechanism for the α1GlyR, suggesting a structural explanation for the effect of some key hyperekplexic allosteric mutations. The first X-ray structure of the TMD of the α1GlyR solved here using GLIC as a scaffold paves the way for mechanistic investigation and design of allosteric modulators of a human receptor.

  11. Allosteric and hyperekplexic mutant phenotypes investigated on an α1 glycine receptor transmembrane structure

    PubMed Central

    Moraga-Cid, Gustavo; Sauguet, Ludovic; Huon, Christèle; Malherbe, Laurie; Girard-Blanc, Christine; Petres, Stéphane; Murail, Samuel; Taly, Antoine; Baaden, Marc; Delarue, Marc; Corringer, Pierre-Jean

    2015-01-01

    The glycine receptor (GlyR) is a pentameric ligand-gated ion channel (pLGIC) mediating inhibitory transmission in the nervous system. Its transmembrane domain (TMD) is the target of allosteric modulators such as general anesthetics and ethanol and is a major locus for hyperekplexic congenital mutations altering the allosteric transitions of activation or desensitization. We previously showed that the TMD of the human α1GlyR could be fused to the extracellular domain of GLIC, a bacterial pLGIC, to form a functional chimera called Lily. Here, we overexpress Lily in Schneider 2 insect cells and solve its structure by X-ray crystallography at 3.5 Å resolution. The TMD of the α1GlyR adopts a closed-channel conformation involving a single ring of hydrophobic residues at the center of the pore. Electrophysiological recordings show that the phenotypes of key allosteric mutations of the α1GlyR, scattered all along the pore, are qualitatively preserved in this chimera, including those that confer decreased sensitivity to agonists, constitutive activity, decreased activation kinetics, or increased desensitization kinetics. Combined structural and functional data indicate a pore-opening mechanism for the α1GlyR, suggesting a structural explanation for the effect of some key hyperekplexic allosteric mutations. The first X-ray structure of the TMD of the α1GlyR solved here using GLIC as a scaffold paves the way for mechanistic investigation and design of allosteric modulators of a human receptor. PMID:25730860

  12. Novel selective allosteric and bitopic ligands for the S1P(3) receptor.

    PubMed

    Jo, Euijung; Bhhatarai, Barun; Repetto, Emanuela; Guerrero, Miguel; Riley, Sean; Brown, Steven J; Kohno, Yasushi; Roberts, Edward; Schürer, Stephan C; Rosen, Hugh

    2012-12-21

    Sphingosine 1-phosphate (S1P) is a lysophospholipid signaling molecule that regulates important biological functions, including lymphocyte trafficking and vascular development, by activating G protein-coupled receptors for S1P, namely, S1P(1) through S1P(5). Here, we map the S1P(3) binding pocket with a novel allosteric agonist (CYM-5541), an orthosteric agonist (S1P), and a novel bitopic antagonist (SPM-242). With a combination of site-directed mutagenesis, ligand competition assay, and molecular modeling, we concluded that S1P and CYM-5541 occupy different chemical spaces in the ligand binding pocket of S1P(3). CYM-5541 allowed us to identify an allosteric site where Phe263 is a key gate-keeper residue for its affinity and efficacy. This ligand lacks a polar moiety, and the novel allosteric hydrophobic pocket permits S1P(3) selectivity of CYM-5541 within the highly similar S1P receptor family. However, a novel S1P(3)-selective antagonist, SPM-242, in the S1P(3) pocket occupies the ligand binding spaces of both S1P and CYM-5541, showing its bitopic mode of binding. Therefore, our coordinated approach with biochemical data and molecular modeling, based on our recently published S1P(1) crystal structure data in a highly conserved set of related receptors with a shared ligand, provides a strong basis for the successful optimization of orthosteric, allosteric, and bitopic modulators of S1P(3).

  13. Allosteric modulation of ATP-gated P2X receptor channels

    PubMed Central

    Coddou, Claudio; Stojilkovic, Stanko S.; Huidobro-Toro, J. Pablo

    2013-01-01

    Seven mammalian purinergic receptor subunits, denoted P2X1 to P2X7, and several spliced forms of these subunits have been cloned. When heterologously expressed, these cDNAs encode ATP-gated non-selective cation channels organized as trimers. All activated receptors produce cell depolarization and promote Ca2+ influx through their pores and indirectly by activating voltage-gated calcium channels. However, the biophysical and pharmacological properties of these receptors differ considerably, and the majority of these subunits are also capable of forming heterotrimers with other members of the P2X receptor family, which confers further different properties. These channels have three ATP binding domains, presumably located between neighboring subunits, and occupancy of at least two binding sites is needed for their activation. In addition to the orthosteric binding sites for ATP, these receptors have additional allosteric sites that modulate the agonist action at receptors, including sites for trace metals, protons, neurosteroids, reactive oxygen species and phosphoinositides. The allosteric regulation of P2X receptors is frequently receptor-specific and could be a useful tool to identify P2X members in native tissues and their roles in signaling. The focus of this review is on common and receptor-specific allosteric modulation of P2X receptors and the molecular base accounting for allosteric binding sites. PMID:21639805

  14. Elucidation of a four-site allosteric network in fibroblast growth factor receptor tyrosine kinases

    PubMed Central

    Chen, Huaibin; Marsiglia, William M; Cho, Min-Kyu; Huang, Zhifeng; Deng, Jingjing; Blais, Steven P; Gai, Weiming; Bhattacharya, Shibani; Neubert, Thomas A; Traaseth, Nathaniel J; Mohammadi, Moosa

    2017-01-01

    Receptor tyrosine kinase (RTK) signaling is tightly regulated by protein allostery within the intracellular tyrosine kinase domains. Yet the molecular determinants of allosteric connectivity in tyrosine kinase domain are incompletely understood. By means of structural (X-ray and NMR) and functional characterization of pathogenic gain-of-function mutations affecting the FGF receptor (FGFR) tyrosine kinase domain, we elucidated a long-distance allosteric network composed of four interconnected sites termed the ‘molecular brake’, ‘DFG latch’, ‘A-loop plug’, and ‘αC tether’. The first three sites repress the kinase from adopting an active conformation, whereas the αC tether promotes the active conformation. The skewed design of this four-site allosteric network imposes tight autoinhibition and accounts for the incomplete mimicry of the activated conformation by pathogenic mutations targeting a single site. Based on the structural similarity shared among RTKs, we propose that this allosteric model for FGFR kinases is applicable to other RTKs. DOI: http://dx.doi.org/10.7554/eLife.21137.001 PMID:28166054

  15. Controlling the rate of organic reactions: rational design of allosteric Diels-Alderase ribozymes

    PubMed Central

    Amontov, Sergey; Jäschke, Andres

    2006-01-01

    Allosteric mechanisms are widely used in nature to control the rates of enzymatic reactions, but little is known about RNA catalysts controlled by these principles. The only natural allosteric ribozyme reported to date catalyzes an RNA cleavage reaction, and so do almost all artificial systems. RNA has, however, been shown to accelerate a much wider range of chemical reactions. Here we report that RNA catalysts for organic reactions can be put under the stringent control of effector molecules by straight-forward rational design. This approach uses known RNA sequences with catalytic and ligand-binding properties, and exploits weakly conserved sequence elements and available structural information to induce the formation of alternative, catalytically inactive structures. The potential and general applicability is demonstrated by the design of three different systems in which the rate of a catalytic carbon–carbon bond forming reaction is positively regulated up to 2100-fold by theophylline, tobramycin and a specific mRNA sequence, respectively. Although smaller in size than a tRNA, all three ribozymes show typical features of allosteric metabolic enzymes, namely high rate acceleration and tight allosteric regulation. Not only do these findings demonstrate RNA's power as a catalyst, but also highlight on RNA's capabilities as signaling components in regulatory networks. PMID:16990253

  16. Controlling the rate of organic reactions: rational design of allosteric Diels-Alderase ribozymes.

    PubMed

    Amontov, Sergey; Jäschke, Andres

    2006-01-01

    Allosteric mechanisms are widely used in nature to control the rates of enzymatic reactions, but little is known about RNA catalysts controlled by these principles. The only natural allosteric ribozyme reported to date catalyzes an RNA cleavage reaction, and so do almost all artificial systems. RNA has, however, been shown to accelerate a much wider range of chemical reactions. Here we report that RNA catalysts for organic reactions can be put under the stringent control of effector molecules by straight-forward rational design. This approach uses known RNA sequences with catalytic and ligand-binding properties, and exploits weakly conserved sequence elements and available structural information to induce the formation of alternative, catalytically inactive structures. The potential and general applicability is demonstrated by the design of three different systems in which the rate of a catalytic carbon-carbon bond forming reaction is positively regulated up to 2100-fold by theophylline, tobramycin and a specific mRNA sequence, respectively. Although smaller in size than a tRNA, all three ribozymes show typical features of allosteric metabolic enzymes, namely high rate acceleration and tight allosteric regulation. Not only do these findings demonstrate RNA's power as a catalyst, but also highlight on RNA's capabilities as signaling components in regulatory networks.

  17. A novel conserved phosphotyrosine motif in the Drosophila fibroblast growth factor signaling adaptor Dof with a redundant role in signal transmission.

    PubMed

    Csiszar, Agnes; Vogelsang, Elisabeth; Beug, Hartmut; Leptin, Maria

    2010-04-01

    The fibroblast growth factor receptor (FGFR) signals through adaptors constitutively associated with the receptor. In Drosophila melanogaster, the FGFR-specific adaptor protein Downstream-of-FGFR (Dof) becomes phosphorylated upon receptor activation at several tyrosine residues, one of which recruits Corkscrew (Csw), the Drosophila homolog of SHP2, which provides a molecular link to mitogen-activated protein kinase (MAPK) activation. However, the Csw pathway is not the only link from Dof to MAPK. In this study, we identify a novel phosphotyrosine motif present in four copies in Dof and also found in other insect and vertebrate signaling molecules. We show that these motifs are phosphorylated and contribute to FGF signal transduction. They constitute one of three sets of phosphotyrosines that act redundantly in signal transmission: (i) a Csw binding site, (ii) four consensus Grb2 recognition sites, and (iii) four novel tyrosine motifs. We show that Src64B binds to Dof and that Src kinases contribute to FGFR-dependent MAPK activation. Phosphorylation of the novel tyrosine motifs is required for the interaction of Dof with Src64B. Thus, Src64B recruitment to Dof through the novel phosphosites can provide a new link to MAPK activation and other cellular responses. This may give a molecular explanation for the involvement of Src kinases in FGF-dependent developmental events.

  18. FM transmission of video signals beyond of the baseband of a 1-km 62.5 μm MMF at the 850 nm wavelength

    NASA Astrophysics Data System (ADS)

    Kowalczyk, Marcin

    2009-06-01

    The paper presents the first, to the best knowledge of the author, successful transmission of the video signal beyond of the baseband of the multimode fibre at the 850 nm wavelength using the subcarrier multiplexing technology (SCM). The transmission system has been built on the basis of generally available, inexpensive optical and electronic components.

  19. 120 Gbit/s injection-locked homodyne coherent transmission of polarization-multiplexed 64 QAM signals over 150 km.

    PubMed

    Wang, Yixin; Kasai, Keisuke; Yoshida, Masato; Nakazawa, Masataka

    2014-12-15

    We describe an injection-locked 64 QAM homodyne coherent transmission, which is the highest QAM multiplicity realized with an injection locking technique. The frequency locking range of the local oscillator (LO) was as wide as 1 GHz. The phase noise was only 0.2 deg, which is 1/3 of that obtained with our previous OVCO-based OPLL (0.6 deg.). As a result, a 120 Gbit/s polarization-multiplexed 64 QAM signal was successfully transmitted over 150 km with a simple receiver configuration and low DSP complexity.

  20. Practical application of a bidirectional microwave photonic filter: simultaneous transmission of analog TV signals

    NASA Astrophysics Data System (ADS)

    Correa-Mena, Ana Gabriela; Zaldívar-Huerta, Ignacio E.; Abril García, Jose Humberto; García-Juárez, Alejandro; Vera-Marquina, Alicia

    2016-10-01

    A practical application of a bidirectional microwave photonic filter (MPF) to transmit simultaneous analog TV signals coded on microwave carriers is experimentally demonstrated. The frequency response of the bidirectional MPF is obtained by the interaction of an externally modulated multimode laser diode emitting at 1.55 μm associated to the free-spectral range of the optical source, the chromatic dispersion parameter of the optical fiber, as well as the length of the optical link. The filtered microwave bandpass window generated around 2 GHz is used as electrical carrier in order to simultaneously transmit TV signals of 67.25 and 61.25 MHz in both directions. The obtained signal-to-noise ratios for the transmitted signals of 67.25 and 61.25 MHz are 37.62 and 44.77 dB, respectively.

  1. Role asymmetry and code transmission in signaling games: an experimental and computational investigation.

    PubMed

    Moreno, Maggie; Baggio, Giosuè

    2015-07-01

    In signaling games, a sender has private access to a state of affairs and uses a signal to inform a receiver about that state. If no common association of signals and states is initially available, sender and receiver must coordinate to develop one. How do players divide coordination labor? We show experimentally that, if players switch roles at each communication round, coordination labor is shared. However, in games with fixed roles, coordination labor is divided: Receivers adjust their mappings more frequently, whereas senders maintain the initial code, which is transmitted to receivers and becomes the common code. In a series of computer simulations, player and role asymmetry as observed experimentally were accounted for by a model in which the receiver in the first signaling round has a higher chance of adjusting its code than its partner. From this basic division of labor among players, certain properties of role asymmetry, in particular correlations with game complexity, are seen to follow.

  2. 47 CFR 11.51 - EAS code and Attention Signal Transmission requirements.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... section on at least one channel. The Attention signal may be produced from a storage device. Additionally... from a storage device. Additionally, these analog cable systems, digital cable systems, and...

  3. MSK and offset QPSK signal transmissions through nonlinear satellite channels in the presence of intersymbol interference

    NASA Astrophysics Data System (ADS)

    Ekanayake, N.

    1983-10-01

    An expression is derived for the bit error probability of minimum shift keying (MSK) and offset quaternary phase shift keying (OQPSK) signals which have been transmitted through nonlinear satellite channels after bandlimiting. The transponder nonlinearity considered in the paper is of the bandpass type, which exhibits AM/AM and AM/PM conversion effects. The effects of up-link and down-link thermal noise are also taken into consideration in the analysis. The resulting expression for the bit error probability is an infinite series containing double integrals. In order to illustrate the usefulness of the method, the error rates of MSK and OQPSK signals are computed for a Butterworth bandlimiting filter hard limiting transponder channel model. The error rates of MSK and OQPSK signals are compared with the error rate of binary PSK signals for this communication system model.

  4. Optical wireless transmission of 405 nm, 1.45 Gbit/s optical IM/DD-OFDM signals through a 4.8 m underwater channel.

    PubMed

    Nakamura, Kazuhiko; Mizukoshi, Izumi; Hanawa, Masanori

    2015-01-26

    In this paper, we experimentally demonstrate wireless transmission of optical intensity modulation/direct detection-orthogonal frequency division multiplexing (IM/DD-OFDM) signals in an underwater channel using a field programmable gate array based real-time transmitter. The real-time transmission of a 405 nm 1.45 Gbit/s optical OFDM signal through a 4.8 m underwater channel with an error vector magnitude of approximately 10% was successfully achieved.

  5. Efficient Processing of Acoustic Signals for High Rate Information Transmission over Sparse Underwater Channels

    DTIC Science & Technology

    2016-09-02

    real- time implementation. To reduce computational complexity of signal processing and improve performance of data detection, receiver structures that...the fractionally-spaced channel estimators and the short feedforward equalizer filters. Receiver algorithm is applied to real data transmitted at 10...on minimization of the mean-squared error in data symbol estimation. This tap selection method is not optimal because the input signal to the

  6. Millimeter-wave signal generation for a wireless transmission system based on on-chip photonic integrated circuit structures.

    PubMed

    Guzmán, R; Carpintero, G; Gordon, C; Orbe, L

    2016-10-15

    We demonstrate and compare two different photonic-based signal sources for generating the carrier wave in a wireless communication link operating in the millimeter-wave range. The first signal source uses the optical heterodyne technique to generate a 113 GHz carrier wave frequency, while the second employs a different technique based on a pulsed mode-locked source with 100 GHz repetition rate frequency. The two optical sources were fabricated in a multi-project wafer run from an active/passive generic integration platform process using standardized building blocks, including multimode interference reflectors which allow us to define the structures on chip, without the need for cleaved facet mirrors. We highlight the superior performance of the mode-locked sources over an optical heterodyne technique. Error-free transmission was achieved in this experiment.

  7. A small animal PET based on GAPDs and charge signal transmission approach for hybrid PET-MR imaging

    NASA Astrophysics Data System (ADS)

    Kang, Jihoon; Choi, Yong; Hong, Key Jo; Hu, Wei; Jung, Jin Ho; Huh, Yoonsuk; Kim, Byung-Tae

    2011-08-01

    Positron emission tomography (PET) employing Geiger-mode avalanche photodiodes (GAPDs) and charge signal transmission approach was developed for small animal imaging. Animal PET contained 16 LYSO and GAPD detector modules that were arranged in a 70 mm diameter ring with an axial field of view of 13 mm. The GAPDs charge output signals were transmitted to a preamplifier located remotely using 300 cm flexible flat cables. The position decoder circuits (PDCs) were used to multiplex the PET signals from 256 to 4 channels. The outputs of the PDCs were digitized and further-processed in the data acquisition unit. The cross-compatibilities of the PET detectors and MRI were assessed outside and inside the MRI. Experimental studies of the developed full ring PET were performed to examine the spatial resolution and sensitivity. Phantom and mouse images were acquired to examine the imaging performance. The mean energy and time resolution of the PET detector were 17.6% and 1.5 ns, respectively. No obvious degradation on PET and MRI was observed during simultaneous PET-MRI data acquisition. The measured spatial resolution and sensitivity at the CFOV were 2.8 mm and 0.7%, respectively. In addition, a 3 mm diameter line source was clearly resolved in the hot-sphere phantom images. The reconstructed transaxial PET images of the mouse brain and tumor displaying the glucose metabolism patterns were imaged well. These results demonstrate GAPD and the charge signal transmission approach can allow the development of high performance small animal PET with improved MR compatibility.

  8. Limits of the propagation medium with respect to digital signal transmission

    NASA Astrophysics Data System (ADS)

    Lampert, E. W.

    1984-10-01

    The effects of the propagation medium are more pronounced in digital communication systems than in analogue ones, in particular when considering single channel systems. The quality parameters: bit-error-rate and link availability are affected not only by the received signal level but also by multipath propagation which may result in frequency selective fading or just in a too low signal level when changing the location in a mobile system or signal fading when the medium is time variable. While fading decreases the signal to noise ratio of links causing short or long terms outages, frequency selective fading causes intersymbol interference and distortion which results in classical systems in an irreducible error rate. Propagation effects for each frequency region from HF to EHF are discussed individually. For LOS-links the typical multipath geometry is presented and data for coherence bandwidth and fading duration are given. Meteor burst scatter can only be used in digital ARQ-systems. It is shown that present systems are rather power limited than distortion limited. In digital troposcatter systems selective fading due to the mutual differential delay of the signal components is a limiting factor especially in digital systems. Delay power spectra are discussed with respect to geometry and layer structure.

  9. Investigating Metabotropic Glutamate Receptor 5 Allosteric Modulator Cooperativity, Affinity, and Agonism: Enriching Structure-Function Studies and Structure-Activity Relationships

    PubMed Central

    Gregory, Karen J.; Noetzel, Meredith J.; Rook, Jerri M.; Vinson, Paige N.; Stauffer, Shaun R.; Rodriguez, Alice L.; Emmitte, Kyle A.; Zhou, Ya; Chun, Aspen C.; Felts, Andrew S.; Chauder, Brian A.; Lindsley, Craig W.; Niswender, Colleen M.

    2012-01-01

    Drug discovery programs increasingly are focusing on allosteric modulators as a means to modify the activity of G protein-coupled receptor (GPCR) targets. Allosteric binding sites are topographically distinct from the endogenous ligand (orthosteric) binding site, which allows for co-occupation of a single receptor with the endogenous ligand and an allosteric modulator that can alter receptor pharmacological characteristics. Negative allosteric modulators (NAMs) inhibit and positive allosteric modulators (PAMs) enhance the affinity and/or efficacy of orthosteric agonists. Established approaches for estimation of affinity and efficacy values for orthosteric ligands are not appropriate for allosteric modulators, and this presents challenges for fully understanding the actions of novel modulators of GPCRs. Metabotropic glutamate receptor 5 (mGlu5) is a family C GPCR for which a large array of allosteric modulators have been identified. We took advantage of the many tools for probing allosteric sites on mGlu5 to validate an operational model of allosterism that allows quantitative estimation of modulator affinity and cooperativity values. Affinity estimates derived from functional assays fit well with affinities measured in radioligand binding experiments for both PAMs and NAMs with diverse chemical scaffolds and varying degrees of cooperativity. We observed modulation bias for PAMs when we compared mGlu5-mediated Ca2+ mobilization and extracellular signal-regulated kinase 1/2 phosphorylation data. Furthermore, we used this model to quantify the effects of mutations that reduce binding or potentiation by PAMs. This model can be applied to PAM and NAM potency curves in combination with maximal fold-shift data to derive reliable estimates of modulator affinities. PMID:22863693

  10. Helicobacter pylori: A Paradigm Pathogen for Subverting Host Cell Signal Transmission.

    PubMed

    Naumann, Michael; Sokolova, Olga; Tegtmeyer, Nicole; Backert, Steffen

    2017-04-01

    Helicobacter pylori colonizes the gastric mucosa in the human stomach and represents a major risk factor for peptic ulcer disease and gastric cancer. Here, we summarize our current knowledge of the complex impact of H. pylori on manipulating host signalling networks, that is, by the cag pathogenicity island (cagPAI)-encoded type IV secretion system (T4SS). We show that H. pylori infections reflect a paradigm for interspecies contact-dependent molecular communication, which includes the disruption of cell-cell junctions and cytoskeletal rearrangements, as well as proinflammatory, cell cycle-related, proliferative, antiapoptotic, and DNA damage responses. The contribution of these altered signalling cascades to disease outcome is discussed.

  11. A system analysis of the 13.3 GHz scatterometer. [antenna patterns and signal transmission

    NASA Technical Reports Server (NTRS)

    Wang, J. R.

    1977-01-01

    The performance of the 13.3 GHz airborne scatterometer system which is used as a microwave remote sensor to detect moisture content of soil is analyzed with respect to its antenna pattern, the signal flow in the receiver data channels, and the errors in the signal outputs. The operational principle and the sensitivity of the system, as well as data handling are also described. The dielectric property of the terrain surface, as far as the scatterometer is concerned, is contained in the assumed forms of the functional dependence of the backscattering coefficient of the incident angle.

  12. Bluetooth telemedicine processor for multichannel biomedical signal transmission via mobile cellular networks.

    PubMed

    Rasid, Mohd Fadlee A; Woodward, Bryan

    2005-03-01

    One of the emerging issues in m-Health is how best to exploit the mobile communications technologies that are now almost globally available. The challenge is to produce a system to transmit a patient's biomedical signals directly to a hospital for monitoring or diagnosis, using an unmodified mobile telephone. The paper focuses on the design of a processor, which samples signals from sensors on the patient. It then transmits digital data over a Bluetooth link to a mobile telephone that uses the General Packet Radio Service. The modular design adopted is intended to provide a "future-proofed" system, whose functionality may be upgraded by modifying the software.

  13. Underwater wireless transmission of high-speed QAM-OFDM signals using a compact red-light laser.

    PubMed

    Xu, Jing; Song, Yuhang; Yu, Xiangyu; Lin, Aobo; Kong, Meiwei; Han, Jun; Deng, Ning

    2016-04-18

    We first study the transmission property of red light in water in terms of extinction coefficient and channel bandwidth via Monte Carlo simulation, with an interesting finding that red light outperforms blue-green light in highly turbid water. We further propose and experimentally demonstrate a broadband underwater wireless optical communication system based on a simple and cost-effective TO56 red-light laser diode. We demonstrate a 1.324-Gb/s transmission at a bit error rate (BER) of 2.02 × 10-3 over a 6-m underwater channel, by using 128-QAM OFDM signals and a low-cost 150-MHz positive-intrinsic-negative photodetector, with a record spectral efficiency higher than 7.32 bits/Hz. By using an avalanche photodetector and 32-QAM OFDM signals, we have achieved a record bit rate of 4.883 Gb/s at a BER of 3.20 × 10-3 over a 6-m underwater channel.

  14. Root gravitropism: an experimental tool to investigate basic cellular and molecular processes underlying mechanosensing and signal transmission in plants

    NASA Technical Reports Server (NTRS)

    Boonsirichai, K.; Guan, C.; Chen, R.; Masson, P. H.

    2002-01-01

    The ability of plant organs to use gravity as a guide for growth, named gravitropism, has been recognized for over two centuries. This growth response to the environment contributes significantly to the upward growth of shoots and the downward growth of roots commonly observed throughout the plant kingdom. Root gravitropism has received a great deal of attention because there is a physical separation between the primary site for gravity sensing, located in the root cap, and the site of differential growth response, located in the elongation zones (EZs). Hence, this system allows identification and characterization of different phases of gravitropism, including gravity perception, signal transduction, signal transmission, and curvature response. Recent studies support some aspects of an old model for gravity sensing, which postulates that root-cap columellar amyloplasts constitute the susceptors for gravity perception. Such studies have also allowed the identification of several molecules that appear to function as second messengers in gravity signal transduction and of potential signal transducers. Auxin has been implicated as a probable component of the signal that carries the gravitropic information between the gravity-sensing cap and the gravity-responding EZs. This has allowed the identification and characterization of important molecular processes underlying auxin transport and response in plants. New molecular models can be elaborated to explain how the gravity signal transduction pathway might regulate the polarity of auxin transport in roots. Further studies are required to test these models, as well as to study the molecular mechanisms underlying a poorly characterized phase of gravitropism that is independent of an auxin gradient.

  15. An engineered chorismate mutase with allosteric regulation.

    PubMed

    Zhang, Sheng; Wilson, David B; Ganem, Bruce

    2003-07-17

    Besides playing a central role in phenylalanine biosynthesis, the bifunctional P-protein in Eschericia coli provides a unique model system for investigating whether allosteric effects can be engineered into protein catalysts using modular regulatory elements. Previous studies have established that the P-protein contains three distinct domains whose functions are preserved, even when separated: chorismate mutase (residues 1-109), prephenate dehydratase (residues 101-285), and an allosteric domain (residues 286-386) for feedback inhibition by phenylalanine. By deleting the prephenate dehydrase domain, a functional chorismate mutase linked directly to the phenylalanine binding domain has been engineered and overexpressed. This manuscript reports the catalytic properties of the mutase in the absence and presence of phenylalanine.

  16. Allosteric inhibition of HIV-1 integrase activity

    PubMed Central

    Engelman, Alan; Kessl, Jacques J.; Kvaratskhelia, Mamuka

    2013-01-01

    HIV-1 integrase is an important therapeutic target in the fight against HIV/AIDS. Integrase strand transfer inhibitors (INSTIs), which target the enzyme active site, have witnessed clinical success over the past 5 years, but the generation of drug resistance poses challenges to INSTI-based therapies moving forward. Integrase is a dynamic protein, and its ordered multimerization is critical to enzyme activity. The integrase tetramer, bound to viral DNA, interacts with host LEDGF/p75 protein to tether integration to active genes. Allosteric integrase inhibitors (ALLINIs) that compete with LEDGF/p75 for binding to integrase disrupt integrase assembly with viral DNA and allosterically inhibit enzyme function. ALLINIs display steep dose response curves and synergize with INSTIs ex vivo, highlighting this novel inhibitor class for clinical development. PMID:23647983

  17. Behavioural innovation and cultural transmission of communication signal in black howler monkeys

    PubMed Central

    Briseño-Jaramillo, M.; Estrada, A.; Lemasson, A.

    2015-01-01

    Social traditions based on communication signals are widespread in birds, cetaceans and humans, but surprisingly rare in nonhuman primates known for having genetically-determined vocal repertoires. This study presents the first description of a singular case of behaviour associated with calling (placing a hand in front of the mouth while vocalizing: HFM) in black howler monkeys. We showed, first, that HFM was found only in a subset of the groups observed, at the same geographical location, and was age- and sex-specific. There was an audience effect on HFM, with highest rates when a neighbouring group was visible. HFM was non-randomly combined with audio-visual signals and always performed while roaring. High HFM rates triggered more vocal responses from group members and male neighbours, and HFM signalers temporally synchronized their behaviour in a predictable way. Finally, the positioning of the hand systematically modified the call’s auditory structure. Altogether these results support the idea that HFM is an innovated, culturally transmitted communication signal that may play a role in inter-group competition and intra-group cohesion. This study opens new lines of research about how nonhuman primates developed strategies to overcome their constraints in acoustic plasticity very early in the primate lineage. PMID:26303965

  18. Floating microbial fuel cells as energy harvesters for signal transmission from natural water bodies

    NASA Astrophysics Data System (ADS)

    Schievano, Andrea; Colombo, Alessandra; Grattieri, Matteo; Trasatti, Stefano P.; Liberale, Alessandro; Tremolada, Paolo; Pino, Claudio; Cristiani, Pierangela

    2017-02-01

    A new type of floating microbial fuel cell (fMFC) was developed for power supply of remote environmental sensors and data transmission. Ten operating fMFCs generated a cell potential in the range 100-800 mV depending on the external resistance applied. Power production peaked around 3-3.5 mW (power density of 22-28 mW m-2 cathode) after about 20-30 days of start-up period. The average of daily electrical energy harvested ranged between 10 and 35 mWh/d. Long-term performances were ensured in the presence of dense rice plants (Oryza Sativa). A power management system, based on a step-up DC/DC converter and a low-power data transmission system via SIGFOX™ technology, have been set up for the fMFCs. The tested fMFCs systems allowed to: i) harvest produced energy, ii) supply electronic devices (intermittent LED-light and a buzzer); iii) transmit remote data at low speed (three message of 12 bites each, in 6 s). Several 'floating garden' MFCs were set in the context of demonstrative events at EXPO2015 world exposition held in Milan between May-October 2015. Some of the 'floating garden' MFCs were operating for more than one year.

  19. Vibrating the food receivers: a direct way of signal transmission in stingless bees (Melipona seminigra).

    PubMed

    Hrncir, Michael; Schmidt, Veronika M; Schorkopf, Dirk Louis P; Jarau, Stefan; Zucchi, Ronaldo; Barth, Friedrich G

    2006-08-01

    An element common to the recruitment communication of eusocial bees (honey bees, stingless bees and bumble bees) are pulsed thorax vibrations generated by successful foragers within the nest. In stingless bees, foragers vibrate during the unloading of the collected food. In the present study on Melipona seminigra we demonstrate that during trophallactic contacts, the food receivers are directly vibrated by the foragers. As a consequence, both the temporal structure and the main frequency component of the forager's vibrations are directly passed on to the receiver. The vibrations are attenuated by about 17 dB on their way from the forager's thorax (velocity amplitude of the vibrations: approximately 70 mm/s) to the receiver's thorax (approximately 10 mm/s), the main amount of attenuation (about 12 dB) occurring during transmission from the head of the forager to that of the receiver. Vibrations conducted through the substrate between the forager and food receiver are comparatively small with velocity amplitudes of 0.3 mm/s. Possible ways of perception and the advantages of vibration transmission by direct contact within the recruitment context are discussed.

  20. Notch1 Regulates Hippocampal Plasticity Through Interaction with the Reelin Pathway, Glutamatergic Transmission and CREB Signaling

    PubMed Central

    Brai, Emanuele; Marathe, Swananda; Astori, Simone; Fredj, Naila Ben; Perry, Elisabeth; Lamy, Christophe; Scotti, Alessandra; Alberi, Lavinia

    2015-01-01

    Notch signaling plays a crucial role in adult brain function such as synaptic plasticity, memory and olfaction. Several reports suggest an involvement of this pathway in neurodegenerative dementia. Yet, to date, the mechanism underlying Notch activity in mature neurons remains unresolved. In this work, we investigate how Notch regulates synaptic potentiation and contributes to the establishment of memory in mice. We observe that Notch1 is a postsynaptic receptor with functional interactions with the Reelin receptor, apolipoprotein E receptor 2 (ApoER2) and the ionotropic receptor, N-methyl-D-aspartate receptor (NMDAR). Targeted loss of Notch1 in the hippocampal CA fields affects Reelin signaling by influencing Dab1 expression and impairs the synaptic potentiation achieved through Reelin stimulation. Further analysis indicates that loss of Notch1 affects the expression and composition of the NMDAR but not AMPAR. Glutamatergic signaling is further compromised through downregulation of CamKII and its secondary and tertiary messengers resulting in reduced cAMP response element-binding (CREB) signaling. Our results identify Notch1 as an important regulator of mechanisms involved in synaptic plasticity and memory formation. These findings emphasize the possible involvement of this signaling receptor in dementia. Highlights In this paper, we propose a mechanism for Notch1-dependent plasticity that likely underlies the function of Notch1 in memory formation: Notch1 interacts with another important developmental pathway, the Reelin cascade. Notch1 regulates both NMDAR expression and composition. Notch1 influences a cascade of cellular events culminating in CREB activation. PMID:26635527

  1. Investigation of allosteric modulation mechanism of metabotropic glutamate receptor 1 by molecular dynamics simulations, free energy and weak interaction analysis

    PubMed Central

    Bai, Qifeng; Yao, Xiaojun

    2016-01-01

    Metabotropic glutamate receptor 1 (mGlu1), which belongs to class C G protein-coupled receptors (GPCRs), can be coupled with G protein to transfer extracellular signal by dimerization and allosteric regulation. Unraveling the dimer packing and allosteric mechanism can be of great help for understanding specific regulatory mechanism and designing more potential negative allosteric modulator (NAM). Here, we report molecular dynamics simulation studies of the modulation mechanism of FITM on the wild type, T815M and Y805A mutants of mGlu1 through weak interaction analysis and free energy calculation. The weak interaction analysis demonstrates that van der Waals (vdW) and hydrogen bonding play an important role on the dimer packing between six cholesterol molecules and mGlu1 as well as the interaction between allosteric sites T815, Y805 and FITM in wild type, T815M and Y805A mutants of mGlu1. Besides, the results of free energy calculations indicate that secondary binding pocket is mainly formed by the residues Thr748, Cys746, Lys811 and Ser735 except for FITM-bound pocket in crystal structure. Our results can not only reveal the dimer packing and allosteric regulation mechanism, but also can supply useful information for the design of potential NAM of mGlu1. PMID:26887338

  2. Investigation of allosteric modulation mechanism of metabotropic glutamate receptor 1 by molecular dynamics simulations, free energy and weak interaction analysis

    NASA Astrophysics Data System (ADS)

    Bai, Qifeng; Yao, Xiaojun

    2016-02-01

    Metabotropic glutamate receptor 1 (mGlu1), which belongs to class C G protein-coupled receptors (GPCRs), can be coupled with G protein to transfer extracellular signal by dimerization and allosteric regulation. Unraveling the dimer packing and allosteric mechanism can be of great help for understanding specific regulatory mechanism and designing more potential negative allosteric modulator (NAM). Here, we report molecular dynamics simulation studies of the modulation mechanism of FITM on the wild type, T815M and Y805A mutants of mGlu1 through weak interaction analysis and free energy calculation. The weak interaction analysis demonstrates that van der Waals (vdW) and hydrogen bonding play an important role on the dimer packing between six cholesterol molecules and mGlu1 as well as the interaction between allosteric sites T815, Y805 and FITM in wild type, T815M and Y805A mutants of mGlu1. Besides, the results of free energy calculations indicate that secondary binding pocket is mainly formed by the residues Thr748, Cys746, Lys811 and Ser735 except for FITM-bound pocket in crystal structure. Our results can not only reveal the dimer packing and allosteric regulation mechanism, but also can supply useful information for the design of potential NAM of mGlu1.

  3. N-Aryl Piperazine Metabotropic Glutamate Receptor 5 Positive Allosteric Modulators Possess Efficacy in Preclinical Models of NMDA Hypofunction and Cognitive Enhancement

    PubMed Central

    Gregory, K.J.; Herman, E.J.; Ramsey, A.J.; Hammond, A.S.; Byun, N.E.; Stauffer, S.R.; Manka, J.T.; Jadhav, S.; Bridges, T.M.; Weaver, C.D.; Niswender, C.M.; Steckler, T.; Drinkenburg, W.H.; Ahnaou, A.; Lavreysen, H.; Macdonald, G.J.; Bartolomé, J.M.; Mackie, C.; Hrupka, B.J.; Caron, M.G.; Daigle, T.L.; Lindsley, C.W.; Conn, P.J.

    2013-01-01

    Impaired transmission through glutamatergic circuits has been postulated to play a role in the underlying pathophysiology of schizophrenia. Furthermore, inhibition of the N-methyl-d-aspartate (NMDA) subtype of ionotropic glutamate receptors (NMDAR) induces a syndrome that recapitulates many of the symptoms observed in patients with schizophrenia. Selective activation of metabotropic glutamate receptor subtype 5 (mGlu5) may provide a novel therapeutic approach for treatment of symptoms associated with schizophrenia through facilitation of transmission through central glutamatergic circuits. Here, we describe the characterization of two novel N-aryl piperazine mGlu5 positive allosteric modulators (PAMs): 2-(4-(2-(benzyloxy)acetyl)piperazin-1-yl)benzonitrile (VU0364289) and 1-(4-(2,4-difluorophenyl)piperazin-1-yl)-2-((4-fluorobenzyl)oxy)ethanone (DPFE). VU0364289 and DPFE induced robust leftward shifts in the glutamate concentration-response curves for Ca2+ mobilization and extracellular signal-regulated kinases 1 and 2 phosphorylation. Both PAMs displayed micromolar affinity for the common mGlu5 allosteric binding site and high selectivity for mGlu5. VU0364289 and DPFE possessed suitable pharmacokinetic properties for dosing in vivo and produced robust dose-related effects in reversing amphetamine-induced hyperlocomotion, a preclinical model predictive of antipsychotic-like activity. In addition, DPFE enhanced acquisition of contextual fear conditioning in rats and reversed behavioral deficits in a mouse model of NMDAR hypofunction. In contrast, DPFE had no effect on reversing apomorphine-induced disruptions of prepulse inhibition of the acoustic startle reflex. These mGlu5 PAMs also increased monoamine levels in the prefrontal cortex, enhanced performance in a hippocampal-mediated memory task, and elicited changes in electroencephalogram dynamics commensurate with procognitive effects. Collectively, these data support and extend the role for the development of novel

  4. THE ANTIPSYCHOTIC POTENTIAL OF MUSCARINIC ALLOSTERIC MODULATION

    PubMed Central

    Bridges, Thomas M.; LeBois, Evan P.; Hopkins, Corey R.; Wood, Michael R.; Jones, Carrie K.; Conn, P. Jeffrey; Lindsley, Craig W.

    2016-01-01

    SUMMARY The cholinergic hypothesis of schizophrenia emerged over 50 years ago based on clinical observations with both anticholinergics and pan-muscarinic agonists. Not until the 1990s did the cholinergic hypothesis of schizophrenia receive renewed enthusiasm based on clinical data with xanomeline, a muscarinic acetylcholine receptor M1/M4-preferring orthosteric agonist. In a clinical trial with Alzheimer’s patients, xanomeline not only improved cognitive performance, but also reduced psychotic behaviors. This encouraging data spurred a second clinical trial in schizophrenic patients, wherein xanomeline significantly improved the positive, negative and cognitive symptom clusters. However, the question remained: Was the antipsychotic efficacy due to activation of M1, M4 or both M1/M4? Classical orthosteric ligands lacked the muscarinic receptor subtype selectivity required to address this key question. More recently, functional assays have allowed for the discovery of ligands that bind at allosteric sites, binding sites distinct from the orthosteric (acetylcholine) site, which are structurally less conserved and thereby afford high levels of receptor subtype selectivity. Recently, allosteric ligands, with unprecedented selectivity for either M1 or M4, have been discovered and have demonstrated comparable efficacy to xanomeline in preclinical antipsychotic and cognition models. These data suggest that selective allosteric activation of either M1 or M4 has antipsychotic potential through distinct, yet complimentary mechanisms. PMID:20520852

  5. Discovery, synthesis, and molecular pharmacology of selective positive allosteric modulators of the δ-opioid receptor.

    PubMed

    Burford, Neil T; Livingston, Kathryn E; Canals, Meritxell; Ryan, Molly R; Budenholzer, Lauren M L; Han, Ying; Shang, Yi; Herbst, John J; O'Connell, Jonathan; Banks, Martyn; Zhang, Litao; Filizola, Marta; Bassoni, Daniel L; Wehrman, Tom S; Christopoulos, Arthur; Traynor, John R; Gerritz, Samuel W; Alt, Andrew

    2015-05-28

    Allosteric modulators of G protein-coupled receptors (GPCRs) have a number of potential advantages compared to agonists or antagonists that bind to the orthosteric site of the receptor. These include the potential for receptor selectivity, maintenance of the temporal and spatial fidelity of signaling in vivo, the ceiling effect of the allosteric cooperativity which may prevent overdose issues, and engendering bias by differentially modulating distinct signaling pathways. Here we describe the discovery, synthesis, and molecular pharmacology of δ-opioid receptor-selective positive allosteric modulators (δ PAMs). These δ PAMs increase the affinity and/or efficacy of the orthosteric agonists leu-enkephalin, SNC80 and TAN67, as measured by receptor binding, G protein activation, β-arrestin recruitment, adenylyl cyclase inhibition, and extracellular signal-regulated kinases (ERK) activation. As such, these compounds are useful pharmacological tools to probe the molecular pharmacology of the δ receptor and to explore the therapeutic potential of δ PAMs in diseases such as chronic pain and depression.

  6. Exploiting protein flexibility to predict the location of allosteric sites

    PubMed Central

    2012-01-01

    Background Allostery is one of the most powerful and common ways of regulation of protein activity. However, for most allosteric proteins identified to date the mechanistic details of allosteric modulation are not yet well understood. Uncovering common mechanistic patterns underlying allostery would allow not only a better academic understanding of the phenomena, but it would also streamline the design of novel therapeutic solutions. This relatively unexplored therapeutic potential and the putative advantages of allosteric drugs over classical active-site inhibitors fuel the attention allosteric-drug research is receiving at present. A first step to harness the regulatory potential and versatility of allosteric sites, in the context of drug-discovery and design, would be to detect or predict their presence and location. In this article, we describe a simple computational approach, based on the effect allosteric ligands exert on protein flexibility upon binding, to predict the existence and position of allosteric sites on a given protein structure. Results By querying the literature and a recently available database of allosteric sites, we gathered 213 allosteric proteins with structural information that we further filtered into a non-redundant set of 91 proteins. We performed normal-mode analysis and observed significant changes in protein flexibility upon allosteric-ligand binding in 70% of the cases. These results agree with the current view that allosteric mechanisms are in many cases governed by changes in protein dynamics caused by ligand binding. Furthermore, we implemented an approach that achieves 65% positive predictive value in identifying allosteric sites within the set of predicted cavities of a protein (stricter parameters set, 0.22 sensitivity), by combining the current analysis on dynamics with previous results on structural conservation of allosteric sites. We also analyzed four biological examples in detail, revealing that this simple coarse

  7. Full-duplex bidirectional transmission of 10-Gb/s millimeter-wave QPSK signal in E-band optical wireless link.

    PubMed

    Fang, Yuan; Yu, Jianjun; Chi, Nan; Xiao, Jiangnan

    2014-01-27

    We experimentally demonstrated full-duplex bidirectional transmission of 10-Gb/s millimeter-wave (mm-wave) quadrature phase shift keying (QPSK) signal in E-band (71-76 GHz and 81-86 GHz) optical wireless link. Single-mode fibers (SMF) are connected at both sides of the antenna for uplink and downlink which realize 40-km SMF and 2-m wireless link for bidirectional transmission simultaneously. We utilized multi-level modulation format and coherent detection in such E-band optical wireless link for the first time. Mm-wave QPSK signal is generated by photonic technique to increase spectrum efficiency and received signal is coherently detected to improve receiver sensitivity. After the coherent detection, digital signal processing is utilized to compensate impairments of devices and transmission link.

  8. The neurotoxic effects of N-methyl-N-nitrosourea on the electrophysiological property and visual signal transmission of rat's retina

    SciTech Connect

    Tao, Ye; Chen, Tao; Liu, Bei; Yang, Guo Qing; Peng, Guanghua; Zhang, Hua; Huang, Yi Fei

    2015-07-01

    The neurotoxic effects of N-methyl-N-nitrosourea (MNU) on the inner retinal neurons and related visual signal circuits have not been described in any animal models or human, despite ample morphological evidences about the MNU induced photoreceptor (PR) degeneration. With the helping of MEA (multielectrode array) recording system, we gained the opportunity to systemically explore the neural activities and visual signal pathways of MNU administrated rats. Our MEA research identified remarkable alterations in the electrophysiological properties and firstly provided instructive information about the neurotoxicity of MNU that affects the signal transmission in the inner retina. Moreover, the spatial electrophysiological functions of retina were monitored and found that the focal PRs had different vulnerabilities to the MNU. The MNU-induced PR dysfunction exhibited a distinct spatial- and time-dependent progression. In contrast, the spiking activities of both central and peripheral RGCs altered synchronously in response to the MNU administration. Pharmacological tests suggested that gap junctions played a pivotal role in this homogeneous response of RGCs. SNR analysis of MNU treated retina suggested that the signaling efficiency and fidelity of inner retinal circuits have been ruined by this toxicant, although the microstructure of the inner retina seemed relatively consolidated. The present study provided an appropriate example of MEA investigations on the toxicant induced pathological models and the effects of the pharmacological compounds on neuron activities. The positional MEA information would enrich our knowledge about the pathology of MNU induced RP models, and eventually be instrumental for elucidating the underlying mechanism of human RP. - Highlights: • We systemically explored the neural activities and visual signal pathways of MNU administrated retinas. • The focal photoreceptors had different vulnerabilities to the MNU administration.

  9. [The role of the ventral nucleus of the lateral lemniscus in sound signal processing and auditory ascending transmission].

    PubMed

    Liu, Hui-Hua; Luo, Feng; Wang, Xin

    2014-06-25

    The ventral nucleus of the lateral lemniscus (VNLL) is an important nucleus in the central auditory pathway which connects the lower brainstem and the midbrain inferior colliculus (IC). Previous studies have demonstrated that neurons in the VNLL could respond to sound signal parameters. Frequency tuning curves (FTCs) of VNLL neurons are generally wider than FTCs of IC neurons, suggesting that the VNLL does not enhance abilities of frequency discrimination and coding. Two types of rate-intensity functions (RIFs) are found in the VNLL: monotonic and non-monotonic RIFs. Intensity-tuning of VNLL neurons are affected by the temporal firing patterns during processing and encoding intensity. There are multiple temporal firing patterns in VNLL neurons. Onset pattern has a precise timing characteristic which is well suited to encode temporal features of stimuli, and also very important to animal behavior including bat's echolocation. The VNLL accepts inputs from lower nuclei, uploads glycine inhibitory outputs to IC, and modulates response characteristics generating and acoustic signal processing of IC neurons. Recent research suggests that fast inhibitory projection from the VNLL may delay the first spike latency of IC neurons, and the delayed inhibitory projection from the VNLL may mediate the temporal firing patterns of IC neurons. But how inhibitory inputs from the VNLL integrate in IC, and how inhibitory inputs from the VNLL enhance the ability of detecting sound signal of IC neurons are not very clear and need more direct evidence at the level of neurons. These questions will help further understand the role of upload during IC processes acoustic signal, which are our research target in the future. This article reviews the current literature regarding the roles of the VNLL in sound signal processing and the auditory ascending transmission, including advances in the relevant research in our laboratory.

  10. Using pulse width modulation for wireless transmission of neural signals in multichannel neural recording systems.

    PubMed

    Yin, Ming; Ghovanloo, Maysam

    2009-08-01

    We have used a well-known technique in wireless communication, pulse width modulation (PWM) of time division multiplexed (TDM) signals, within the architecture of a novel wireless integrated neural recording (WINeR) system. We have evaluated the performance of the PWM-based architecture and indicated its accuracy and potential sources of error through detailed theoretical analysis, simulations, and measurements on a setup consisting of a 15-channel WINeR prototype as the transmitter and two types of receivers; an Agilent 89600 vector signal analyzer and a custom wideband receiver, with 36 and 75 MHz of maximum bandwidth, respectively. Furthermore, we present simulation results from a realistic MATLAB-Simulink model of the entire WINeR system to observe the system behavior in response to changes in various parameters. We have concluded that the 15-ch WINeR prototype, which is fabricated in a 0.5- mum standard CMOS process and consumes 4.5 mW from +/-1.5 V supplies, can acquire and wirelessly transmit up to 320 k-samples/s to a 75-MHz receiver with 8.4 bits of resolution, which is equivalent to a wireless data rate of approximately 2.56 Mb/s.

  11. Transmission and pass-drop operations of mixed baudrate Nyquist OTDM-WDM signals for all-optical elastic network.

    PubMed

    Tan, Hung Nguyen; Inoue, Takashi; Kurosu, Takayuki; Namiki, Shu

    2013-08-26

    We propose the use of Nyquist OTDM-WDM signal for highly efficient, fully elastic all-optical networks. With the possibility of generation of ultra-coarse yet flexible granular channels, Nyquist OTDM-WDM can eliminate guard-bands in conventional WDM systems, and hence improves the spectral efficiency in network perspective. In this paper, transmission and pass-drop operations of mixed baudrate Nyquist OTDM-WDM channels from 43 Gbaud to dual-polarization 344 Gbaud are successfully demonstrated over 320 km fiber link with four FlexGrid-compatible WSS nodes. A stable clock recovery is also carried out for different baudrate Nyquist OTDMs by optical null-header insertion technique.

  12. InP-based photonic integrated circuits for optical performance surveillance, signal conditioning, and bandwidth management in DWDM transmission systems

    NASA Astrophysics Data System (ADS)

    Tolstikhin, Valery I.; Wu, Fang; Logvin, Yury; Densmore, Adam; Pimenov, Kirill; Grabtchak, Serge

    2004-11-01

    This paper reports the design of InP-based monolithic photonic integrated circuits for performance surveillance and bandwidth management in DWDM transmission systems. It is based on a building block approach, which allows a large variety of optical components to be built from a few monolithically integrable elements, by using only one-step epitaxial growth and standard semiconductor fabrication technologies. These include: (i) polarization-compensated echelle diffractive grating (de)multiplexer, along with the elements of passive waveguide circuitry for coupling the light to and directing it through the InP-based photonic chip, and (ii) single-mode vertically integrated waveguide active devices with detecting, attenuating and amplifying features, inserted in the (in)output channels of the planar (de)multiplexer. The paper presents the design and characterization examples of these elements and discusses the related integrated components for controlling / manipulating the DWDM optical signals on a per frequency basis.

  13. Transmissive grating-reflective mirror-based fiber optic accelerometer for stable signal acquisition in industrial applications

    NASA Astrophysics Data System (ADS)

    Lee, Yeon-Gwan; Kim, Dae-Hyun; Kim, Chun-Gon

    2012-05-01

    This paper discusses an applicable fiber-optic accelerometer composed of a transmissive grating panel, a reflection mirror, and two optical fibers with a separation of quarter grating pitch as transceivers that monitor the low-frequency accelerations of civil engineering structures. This sensor structure brings together the advantages of both a simple sensor structure, which leads to simplified cable design by 50% in comparison with the conventional transmission-type fiber optic accelerometer, and a stable reflected signals acquisition with repeatability in comparison to the researched grating-reflection type fiber optic accelerometer. The vibrating displacement and sinusoidal acceleration measured from the proposed fiber optic sensor demonstrated good agreement with those of a commercial laser displacement sensor and a MEMS accelerometer without electromagnetic interference. The developed fiber optic accelerometer can be used in frequency ranges below 4.0 Hz with a margin of error that is less than 5% and a high sensitivity of 5.06 rad/(m/s)2.

  14. Diffusible signal factor-repressed extracellular traits enable attachment of Xylella fastidiosa to insect vectors and transmission.

    PubMed

    Baccari, Clelia; Killiny, Nabil; Ionescu, Michael; Almeida, Rodrigo P P; Lindow, Steven E

    2014-01-01

    The hypothesis that a wild-type strain of Xylella fastidiosa would restore the ability of rpfF mutants blocked in diffusible signal factor production to be transmitted to new grape plants by the sharpshooter vector Graphocephala atropunctata was tested. While the rpfF mutant was very poorly transmitted by vectors irrespective of whether they had also fed on plants infected with the wild-type strain, wild-type strains were not efficiently transmitted if vectors had fed on plants infected with the rpfF mutant. About 100-fewer cells of a wild-type strain attached to wings of a vector when suspended in xylem sap from plants infected with an rpfF mutant than in sap from uninfected grapes. The frequency of transmission of cells suspended in sap from plants that were infected by the rpfF mutant was also reduced over threefold. Wild-type cells suspended in a culture supernatant of an rpfF mutant also exhibited 10-fold less adherence to wings than when suspended in uninoculated culture media. A factor released into the xylem by rpfF mutants, and to a lesser extent by the wild-type strain, thus inhibits their attachment to, and thus transmission by, sharpshooter vectors and may also enable them to move more readily through host plants.

  15. Heterodyne detection and transmission of 60-Gbaud PDM-QPSK signal with SE of 4b/s/Hz.

    PubMed

    Li, Xinying; Xiao, Jiangnan; Yu, Jianjun

    2014-04-21

    We experimentally demonstrate 8 × 240-Gb/s super-Nyquist wavelength-division-multiplexing (WDM) polarization-division-multiplexing quadrature-phase-shift-keying (PDM-QPSK) signal transmission on a 50-GHz grid with a net spectral efficiency (SE) of 4b/s/Hz adopting hardware-efficient simplified heterodyne detection. 9-ary quadrature-amplitude-modulation-like (9QAM-like) processing based on multi-modulus blind equalization (MMBE) is adopted to reduce analog-to-digital converter (ADC) bandwidth requirement and improve receiver sensitivity. The transmission distance at the soft-decision forward-error-correction (SD-FEC) threshold of 2 × 10(-2) is 2 × 420 km based on digital post filtering while largely extended to over 5 × 420 km based on 9QAM-like processing, which well illustrates 9QAM-like processing is more efficient for heterodyne coherent WDM system. Moreover, only two ADC channels are needed for simplified heterodyne detection of one 60-Gbaud PDM-QPSK WDM channel, and thus only one commercial oscilloscope (OSC) with two input ports can work well for each WDM channel.

  16. Transmission of 40-Gb/s QPSK upstream signal in RSOA-based coherent WDM PON using offset PDM technique.

    PubMed

    Shim, H K; Cho, K Y; Hong, U H; Chung, Y C

    2013-02-11

    We demonstrate the 40-Gb/s upstream transmission in the 60-km reach wavelength-division-multiplexed passive optical network (WDM PON) implemented by using directly modulated reflective semiconductor optical amplifiers (RSOAs) and self-homodyne receivers. It is difficult to operate the RSOA at 40 Gb/s due to its limited modulation bandwidth. To overcome this problem and generate 40-Gb/s upstream signal, we utilize the quadrature phase-shift-keying (QPSK) format and the offset polarization-division-multiplexing (PDM) technique. For this purpose, we install two RSOAs at each ONU and provide the seed light for these RSOAs by polarization-multiplexing the outputs of two lasers with a small frequency offset (20 GHz). This frequency offset is used to separate the polarization-multiplexed seed light by using a simple delay-line interferometer (DLI), instead of the polarization-beam splitter and polarization controller, at the ONU. The separated seed light is modulated by each RSOA at 20 Gb/s in the QPSK format, and then combined again by the DLI before sent back to the central office (CO). The results show that this WDM PON can support the transmission of 40-Gb/s channels spaced at 50 GHz over 60 km without using any remote optical amplifiers.

  17. Transmission of wireless neural signals through a 0.18 µm CMOS low-power amplifier.

    PubMed

    Gazziro, M; Braga, C F R; Moreira, D A; Carvalho, A C P L F; Rodrigues, J F; Navarro, J S; Ardila, J C M; Mioni, D P; Pessatti, M; Fabbro, P; Freewin, C; Saddow, S E

    2015-01-01

    In the field of Brain Machine Interfaces (BMI) researchers still are not able to produce clinically viable solutions that meet the requirements of long-term operation without the use of wires or batteries. Another problem is neural compatibility with the electrode probes. One of the possible ways of approaching these problems is the use of semiconductor biocompatible materials (silicon carbide) combined with an integrated circuit designed to operate with low power consumption. This paper describes a low-power neural signal amplifier chip, named Cortex, fabricated using 0.18 μm CMOS process technology with all electronics integrated in an area of 0.40 mm(2). The chip has 4 channels, total power consumption of only 144 μW, and is impedance matched to silicon carbide biocompatible electrodes.

  18. Maternal-Fetal Transmission of Zika Virus: Routes and Signals for Infection.

    PubMed

    Cao, Bin; Diamond, Michael S; Mysorekar, Indira U

    2017-04-12

    The emerging mosquito-borne virus, Zika virus (ZIKV), has been causally associated with adverse pregnancy and neonatal outcomes, including miscarriage, microcephaly, serious brain abnormalities, and other birth defects indicative of a congenital ZIKV syndrome. In this review, we highlight work from human and animal studies on routes of infection in pregnancy that lead to adverse fetal and neonatal outcomes. A number of innate and adaptive immune mechanisms and signaling molecules that may have key roles in ZIKV infection pathogenesis are discussed along with putative viral entry pathways. A more granular understanding of pathogenesis of ZIKV infection during pregnancy is critical for developing therapeutics and vaccines and mounting a global public health response to limit ZIKV infections. We also report on new therapeutic interventions that have shown success in preclinical studies.

  19. [Molecular mechanisms for signal-transmission of mechanical stress into bone-forming cells].

    PubMed

    Hakeda, Yoshiyuki; Kumegawa, Masayoshi

    2003-04-01

    One function of bone in organism is to mechanically support the body. The bone is always exposed to mechanical stress such as gravity and locomotion, and the shape of bone is adapted to the mechanical loading. Mechanical loading on bone generates extracellular deformation and fluid flow, and the mechanical stimuli are translated to mechanical signals such as mechanical strain and fluid shear stress. Bone-forming cells such as osteocytes and osteoblasts are mechanosensors. When these cells receive the mechanical stress, they stimulate the production of local regulators for bone metabolism such as prostaglandins, and various growth factors and cytokines. By the actions of these factors on bone-forming cells and bone-resorbing cells in bone microenvironment, the bone metabolism is turn over in conformity with the mechanical stress.

  20. Collective retention and transmission of chemical signals in a social insect

    NASA Astrophysics Data System (ADS)

    Gill, Katherine P.; van Wilgenburg, Ellen; Taylor, Peter; Elgar, Mark A.

    2012-03-01

    Social insect colonies exhibit highly coordinated responses to ecological challenges by acquiring information that is disseminated throughout the colony. Some responses are coordinated directly from the signals produced by individuals that acquired the information. Other responses may require information to be transferred indirectly through a third party, thereby requiring colony-wide retention of information. Social insects use colony signature odours to distinguish between nestmates and non-nestmates, and the level of aggression between non-nestmates typically varies according to the distance between colonies and thus their history of interactions. Such coordinated, colony-specific responses may require information about particular odours to be disseminated and retained across the colony. Our field experiments with weaver ants reveal colony-wide, indirect acquisition and retention of the signature odours of a different colony with which they had experienced aggression. These data highlight the significance of interaction history and suggest the presence of a collective memory.

  1. Allosteric ACTion: the varied ACT domains regulating enzymes of amino-acid metabolism.

    PubMed

    Lang, Eric J M; Cross, Penelope J; Mittelstädt, Gerd; Jameson, Geoffrey B; Parker, Emily J

    2014-12-01

    Allosteric regulation of enzyme activity plays important metabolic roles. Here we review the allostery of enzymes of amino-acid metabolism conferred by a discrete domain known as the ACT domain. This domain of 60-70 residues has a βαββαβ topology leading to a four-stranded β4β1β3β2 antiparallel sheet with two antiparallel helices on one face. Extensive sequence variation requires a combined sequence/structure/function analysis for identification of the ACT domain. Common features include highly varied modes of self-association of ACT domains, ligand binding at domain interfaces, and transmittal of allosteric signals through conformational changes and/or the manipulation of quaternary equilibria. A recent example illustrates the relatively facile adoption of this versatile module of allostery by gene fusion.

  2. Lipid-Mediated Regulation of Embedded Receptor Kinases via Parallel Allosteric Relays.

    PubMed

    Ghosh, Madhubrata; Wang, Loo Chien; Ramesh, Ranita; Morgan, Leslie K; Kenney, Linda J; Anand, Ganesh S

    2017-02-28

    Membrane-anchored receptors are essential cellular signaling elements for stimulus sensing, propagation, and transmission inside cells. However, the contributions of lipid interactions to the function and dynamics of embedded receptor kinases have not been described in detail. In this study, we used amide hydrogen/deuterium exchange mass spectrometry, a sensitive biophysical approach, to probe the dynamics of a membrane-embedded receptor kinase, EnvZ, together with functional assays to describe the role of lipids in receptor kinase function. Our results reveal that lipids play an important role in regulating receptor function through interactions with transmembrane segments, as well as through peripheral interactions with nonembedded domains. Specifically, the lipid membrane allosterically modulates the activity of the embedded kinase by altering the dynamics of a glycine-rich motif that is critical for phosphotransfer from ATP. This allostery in EnvZ is independent of membrane composition and involves direct interactions with transmembrane and periplasmic segments, as well as peripheral interactions with nonembedded domains of the protein. In the absence of the membrane-spanning regions, lipid allostery is propagated entirely through peripheral interactions. Whereas lipid allostery impacts the phosphotransferase function of the kinase, extracellular stimulus recognition is mediated via a four-helix bundle subdomain located in the cytoplasm, which functions as the osmosensing core through osmolality-dependent helical stabilization. Our findings emphasize the functional modularity in a membrane-embedded kinase, separated into membrane association, phosphotransferase function, and stimulus recognition. These components are integrated through long-range communication relays, with lipids playing an essential role in regulation.

  3. Transmission and full-band coherent detection of polarization-multiplexed all-optical Nyquist signals generated by Sinc-shaped Nyquist pulses.

    PubMed

    Zhang, Junwen; Yu, Jianjun; Chi, Nan

    2015-09-01

    All optical method is considered as a promising technique for high symbol rate Nyquist signal generation, which has attracted a lot of research interests for high spectral-efficiency and high-capacity optical communication system. In this paper, we extend our previous work and report the fully experimental demonstration of polarization-division multiplexed (PDM) all-optical Nyquist signal generation based on Sinc-shaped Nyquist pulse with advanced modulation formats, fiber-transmission and single-receiver full-band coherent detection. Using this scheme, we have successfully demonstrated the generation, fiber transmission and single-receiver full-band coherent detection of all-optical Nyquist PDM-QPSK and PDM-16QAM signals up to 125-GBaud. 1-Tb/s single-carrier PDM-16QAM signal generation and full-band coherent detection is realized, which shows the advantage and feasibility of the single-carrier all-optical Nyquist signals.

  4. Transmission and full-band coherent detection of polarization-multiplexed all-optical Nyquist signals generated by Sinc-shaped Nyquist pulses

    PubMed Central

    Zhang, Junwen; Yu, Jianjun; Chi, Nan

    2015-01-01

    All optical method is considered as a promising technique for high symbol rate Nyquist signal generation, which has attracted a lot of research interests for high spectral-efficiency and high-capacity optical communication system. In this paper, we extend our previous work and report the fully experimental demonstration of polarization-division multiplexed (PDM) all-optical Nyquist signal generation based on Sinc-shaped Nyquist pulse with advanced modulation formats, fiber-transmission and single-receiver full-band coherent detection. Using this scheme, we have successfully demonstrated the generation, fiber transmission and single-receiver full-band coherent detection of all-optical Nyquist PDM-QPSK and PDM-16QAM signals up to 125-GBaud. 1-Tb/s single-carrier PDM-16QAM signal generation and full-band coherent detection is realized, which shows the advantage and feasibility of the single-carrier all-optical Nyquist signals. PMID:26323238

  5. Transmission and full-band coherent detection of polarization-multiplexed all-optical Nyquist signals generated by Sinc-shaped Nyquist pulses

    NASA Astrophysics Data System (ADS)

    Zhang, Junwen; Yu, Jianjun; Chi, Nan

    2015-09-01

    All optical method is considered as a promising technique for high symbol rate Nyquist signal generation, which has attracted a lot of research interests for high spectral-efficiency and high-capacity optical communication system. In this paper, we extend our previous work and report the fully experimental demonstration of polarization-division multiplexed (PDM) all-optical Nyquist signal generation based on Sinc-shaped Nyquist pulse with advanced modulation formats, fiber-transmission and single-receiver full-band coherent detection. Using this scheme, we have successfully demonstrated the generation, fiber transmission and single-receiver full-band coherent detection of all-optical Nyquist PDM-QPSK and PDM-16QAM signals up to 125-GBaud. 1-Tb/s single-carrier PDM-16QAM signal generation and full-band coherent detection is realized, which shows the advantage and feasibility of the single-carrier all-optical Nyquist signals.

  6. Noise-driven signal transmission device using molecular dynamics of organic polymers

    NASA Astrophysics Data System (ADS)

    Asakawa, Naoki; Umemura, Koichiro; Fujise, Shinya; Yazawa, Koji; Shimizu, Tadashi; Tansho, Masataka; Kanki, Teruo; Tanaka, Hidekazu

    2014-01-01

    Stochastic threshold devices using a trap-filling transition (TFT) coupled with molecular dynamics in poly(3-alkylthiophene)s were fabricated as potential key devices for noise-driven bioinspired sensors and information processors. This article deals with variable-temperature direct current conductivity and alternating current impedance measurements for vertical-type device elements of Au/regioregular poly(3-decylthiophene) ((RR-P3DT) (thickness: 100 nm)/Au, which show multiple conducting states and quasi-stochastic transitions between these states. Noise measurements indicate the ω-2-type (if VVTFT) power spectral densities, where V and VTFT are an applied voltage and the voltage for TFT, respectively. The noise generation is due to the TFT associated with twist dynamics of π-conjugated polymers near the order-disorder phase transition (ODT). At 298 K, the quasi-stochastic behavior is more noticeable for RR-P3DT than poly(3-hexylthiophene). The quasi-stochastic property is employed to a stochastic one-directional signal transmitting device using optical-electric conversion. The dynamics of ODT for powder samples were also investigated by differential scanning calorimetry measurements and high-resolution solid-state C13 nuclear magnetic resonance spectroscopy, and the correlation of the molecular structure and dynamics with electric properties was discussed.

  7. Asthma families show transmission disequilibrium of gene variants in the vitamin D metabolism and signalling pathway

    PubMed Central

    Wjst, Matthias; Altmüller, Janine; Faus-Kessler, Theresia; Braig, Christine; Bahnweg, Margret; André, Elisabeth

    2006-01-01

    The vitamin D prophylaxis of rickets in pregnant women and newborns may play a role in early allergic sensitization. We now asked if an already diseased population may have inherited genetic variants in the vitamin D turnover or signalling pathway. Serum levels of calcidiol (25-OH-D3) and calcitriol (1,25-(OH)2-D3) were retrospectively assessed in 872 partipants of the German Asthma Family Study. 96 DNA single base variants in 13 different genes were genotyped with MALDI-TOF and a bead array system. At least one positive SNP with a TDT of p < 0.05 for asthma or total IgE and calcidiol or calcitriol was seen in IL10, GC, IL12B, CYP2R1, IL4R, and CYP24A1. Consistent strong genotypic association could not be observed. Haplotype association were found only for CYP24A1, the main calcidiol degrading enzyme, where a frequent 5-point-haplotype was associated with asthma (p = 0,00063), total IgE (p = 0,0014), calcidiol (p = 0,0043) and calcitriol (p = 0,0046). Genetic analysis of biological pathways seem to be a promising approach where this may be a first entry point into effects of a polygenic inherited vitamin D sensitivity that may affect also other metabolic, immunological and cancerous diseases. PMID:16600026

  8. Identification of Allosteric Disulfides from Prestress Analysis

    PubMed Central

    Zhou, Beifei; Baldus, Ilona B.; Li, Wenjin; Edwards, Scott A.; Gräter, Frauke

    2014-01-01

    Disulfide bonds serve to form physical cross-links between residues in protein structures, thereby stabilizing the protein fold. Apart from this purely structural role, they can also be chemically active, participating in redox reactions, and they may even potentially act as allosteric switches controlling protein functions. Specific types of disulfide bonds have been identified in static protein structures from their distinctive pattern of dihedral bond angles, and the allosteric function of such bonds is purported to be related to the torsional strain they store. Using all-atom molecular-dynamics simulations for ∼700 disulfide bonded proteins, we analyzed the intramolecular mechanical forces in 20 classes of disulfide bonds. We found that two particular classes, the −RHStaple and the −/+RHHook disulfides, are indeed more stressed than other disulfide bonds, but the stress is carried primarily by stretching of the S-S bond and bending of the neighboring bond angles, rather than by dihedral torsion. This stress corresponds to a tension force of magnitude ∼200 pN, which is balanced by repulsive van der Waals interactions between the cysteine Cα atoms. We confirm stretching of the S-S bond to be a general feature of the −RHStaples and the −/+RHHooks by analyzing ∼20,000 static protein structures. Given that forced stretching of S-S bonds is known to accelerate their cleavage, we propose that prestress of allosteric disulfide bonds has the potential to alter the reactivity of a disulfide, thereby allowing us to readily switch between functional states. PMID:25099806

  9. Optimization of wide-angle seismic signal-to-noise ratios and P-wave transmission in Kenya

    USGS Publications Warehouse

    Jacob, A.W.B.; Vees, R.; Braile, L.W.; Criley, E.

    1994-01-01

    In previous refraction and wide-angle reflection experiments in the Kenya Rift there were problems with poor signal-noise ratios which made good seismic interpretation difficult. Careful planning and preparation for KRISP 90 has substantially overcome these problems and produced excellent seismic sections in a difficult environment. Noise levels were minimized by working, as far as possible, at times of the day when conditions were quiet, while source signals were optimized by using dispersed charges in water where it was available and waterfilled boreholes in most cases where it was not. Seismic coupling at optimum depth in water has been found to be more than 100 times greater than it is in a borehole in dry loosely compacted material. Allowing for the source coupling, a very marked difference has been found between the observation ranges in the rift and those on the flanks, where the observation ranges are greater. These appear to indicate a significant difference in seismic transmission through the two types of crust. ?? 1994.

  10. Feasibility study of monitoring of plasma etching chamber conditions using superimposed high-frequency signals on rf power transmission line

    NASA Astrophysics Data System (ADS)

    Kasashima, Y.; Uesugi, F.

    2015-10-01

    An in situ monitoring system that can detect changes in the conditions of a plasma etching chamber has been developed. In the system, low-intensity high-frequency signals are superimposed on the rf power transmission line used for generating plasma. The system measures reflected high-frequency signals and detects the change in their frequency characteristics. The results indicate that the system detects the changes in the conditions in etching chambers caused by the changes in the electrode gap and the inner wall condition and demonstrate the effectiveness of the system. The system can easily be retrofitted to mass-production equipment and it can be used with or without plasma discharge. Therefore, our system is suitable for in situ monitoring of mass-production plasma etching chambers. The system is expected to contribute to development of predictive maintenance, which monitors films deposited on the inner wall of the chamber and prevents equipment faults caused by misalignment of chamber parts in mass-production equipment.

  11. Frequency-selective transmission of graded signals in large monopolar neurons of blowfly Calliphora vicina compound eye

    PubMed Central

    Weckström, Matti

    2016-01-01

    The functional roles of voltage-gated K+ (Kv) channels in visual system interneurons remain poorly studied. We have addressed this problem in the large monopolar cells (LMCs) of the blowfly Calliphora vicina, using intracellular recordings and mathematical modeling methods. Intracellular recordings were performed in two cellular compartments: the synaptic zone, which receives input from photoreceptors, and the axon, which provides graded potential output to the third-order visual neurons. Biophysical properties of Kv conductances in the physiological voltage range were examined in the dark with injections of current in the discontinuous current-clamp mode. Putative LMC types 1/2 and 3 (L1/2 and L3, respectively) had dissimilar Kv channelomes: L1/2 displayed a prominent inactivating Kv conductance in the axon, while L3 cells were characterized by a sustained delayed-rectifier Kv conductance. To study the propagation of voltage signals, the data were incorporated into the previously developed mathematical model. We demonstrate that the complex interaction between the passive membrane properties, Kv conductances, and the neuronal geometry leads to a resonance-like filtering of signals with peak frequencies of transmission near 15 and 40 Hz for L3 and L1/2, respectively. These results point to distinct physiological roles of different types of LMCs. PMID:26843598

  12. Retinal signal transmission in Duchenne muscular dystrophy: evidence for dysfunction in the photoreceptor/depolarizing bipolar cell pathway.

    PubMed Central

    Fitzgerald, K M; Cibis, G W; Giambrone, S A; Harris, D J

    1994-01-01

    There have been reports of abnormal retinal neurotransmission determined by electroretinography in boys with Duchenne and Becker muscular dystrophy. Dystrophin may play a role in transmitting signals between photoreceptors and the excitatory synapse of the ON-bipolar cell. These electroretinographic changes appeared to be limited to the rod ON-pathway but we felt there was also similar abnormality in the cone ON-pathway. We used long-duration stimuli to separate ON-(depolarizing bipolar cell) and OFF (hyperpolarizing bipolar cell) contributions to the cone-dominated ERG to better understand how the retina functions in boys with Duchenne muscular dystrophy. We recorded the electroretinograms of 11 boys with Duchenne muscular dystrophy and found abnormal signal transmission at the level of the photoreceptor and ON-bipolar cell in both the rod and cone generated responses. The OFF-bipolar cell that responds to the offset of the stimulus continues to function normally. The results support our hypothesis that retinal dystrophin plays a role in receptor function or controlling ion channels at the level of the photoreceptor and depolarizing bipolar cell. PMID:8200977

  13. Targeting allosteric disulphide bonds in cancer.

    PubMed

    Hogg, Philip J

    2013-06-01

    Protein action in nature is generally controlled by the amount of protein produced and by chemical modification of the protein, and both are often perturbed in cancer. The amino acid side chains and the peptide and disulphide bonds that bind the polypeptide backbone can be post-translationally modified. Post-translational cleavage or the formation of disulphide bonds are now being identified in cancer-related proteins and it is timely to consider how these allosteric bonds could be targeted for new therapies.

  14. Radiation quality dependence of signal transmission and bystander induced cell killing

    NASA Astrophysics Data System (ADS)

    Esposito, Giuseppe; Bertolotti, Alessia; Facoetti, Angelica; Grande, Sveva; Mariotti, Luca; Ottolenghi, Andrea; Ranza, Elena; Simone, Giustina; Sorrentino, Eugenio; Antonella Tabocchini, Maria

    at 20 hours post exposure, whereas IL-8 release was significantly increased at shorter times, i.e. 5-7 hours after irradiation. The expression of their receptors was modulated in both irradiated and bystander cells, although it is not apparently correlated with the relative interleukin re-lease. In order to investigate possible correlation between NRS and cytokines as early and late mediators in the signalling chain leading to bystander induced cell killing, experiments were performed using c-PTIO, a well known scavengers of RNS. In these experiments conditioned medium taken after 1h or 5h from α-particle irradiated cells was used. The results obtained after 5h in the absence of c-PTIO didn't show any further decrease in clonogenic survival of bystander cells. The presence of the scavenger seems to reduce, the bystander induced cell killing indicating that RNS are involved in the transduction of the bystander signal. Experi-ments, performed with both c-PTIO and DMSO, a scavenger of hydroxyl radicals, showed that RNS and ROS play a role in some cytokine pathways (interleukins release and their receptor expression) activated by irradiation. ACKNOWLEDGEMENTS This work was partially supported by the European Commission (EC Contract FP6-36465, "NOTE")

  15. Anharmonicity, neural-like lattices, and fast signal/electric transmission

    NASA Astrophysics Data System (ADS)

    Velarde, Manuel G.

    2007-02-01

    Anharmonic interactions in lattices may sustain robust oscillatory modes and (nonlinear) waves including solitons. This is illustrated here by using an exponentially repulsive interaction introduced by Toda. To cope with friction and dissipation -always present in real systems- and hence to make robust, e.g., solitons, following Lord Rayleigh, an appropriate input-output energy balance is added to the dynamics. Noise (and hence temperature) is also incorporated by embedding the system in a Gaussian, white noise environment (thermal bath). In the particular case of a lattice ring with six units it is shown how such a Toda-Rayleigh lattice can be used as a Central Pattern Generator of three different oscillatory modes. These three modes are shown to map three walking (metachronal/low speed, caterpillar/medium speed, and tripod/fast speed) gaits in insects (hexapods). An electronic implementation (diodes map easily exponential interactions) of the Toda-Rayleigh lattice ring is also discussed, including leg motor controls for an hexapod robot. Finally, the Toda-Rayleigh mechanical lattice is converted into an electromechanical wire-like, lattice electric conductor. This is done by considering the lattice units as positive ion cores and adding free electrons to the system. The coupling of Toda dynamics with Coulomb interactions yields remarkable current-field/voltage and current-temperature characteristics in the presence of an external electric field. An Ohmic-non Ohmic transition is possible in the lattice conductor. Such feature permits to consider it as a neural-like conveyor of subsonic (Ohmic) and fast supersonic (non-Ohmic) electric or other signals.

  16. 100.29-Gb/s direct detection optical OFDM/OQAM 32-QAM signal over 880  km SSMF transmission using a single photodiode.

    PubMed

    Li, Chao; Yang, Qi; Luo, Ming; He, Zhixue; Li, Haibo; Hu, Rong; Yu, Shaohua

    2015-04-01

    We propose a novel guard-band-shared direct-detection (GBS-DD) scheme for a 100-Gb/s single-photodiode direct-detection transmission system. The 100.29 Gb/s signal is successfully transmitted over 880 km standard single mode fiber (SSMF) with Raman amplification under the 20% forward error correction (FEC) threshold within 52 GHz optical bandwidth. The signal is modulated with orthogonal frequency-division multiplexing/offset quadrature-amplitude-modulation 32-ary QAM (OFDM/OQAM 32-QAM). The signal-to-signal beat interference (SSBI) terms fall and overlap in the same guard band. Adopting the proposed approach, the bandwidth usage efficiency of the photodiode is greatly enhanced, which brings benefits on the data rate and transmission performance.

  17. Pulse-grouping transmission of optical quadrature phase-shift keying signals with time diversity multiple-input and multiple-output detection and processing

    NASA Astrophysics Data System (ADS)

    Gao, Guanjun; Chen, Sai; Zhang, Jie

    2016-08-01

    We investigate an optical pulse-overlap transmission scheme where the orthogonal condition between neighbor pulses is violated. The interferences between the grouped optical pulses are mitigated at the optical coherent receiver with time diversity multiple-input and multiple-output-based digital signal processing. Numerical simulation investigates the performance of 50% return-to-zero (RZ)-quadrature phase-shift keying (QPSK) signals, where up to four pulses are overlapped and grouped for per pulse period. In the experiment demonstration, two 50% RZ-QPSK signals are combined with different time offset between neighbor pulses, and the Q-performance as a function of optical-signal-to-noise ratio (OSNR) is compared on each pulse channel basis, with minimum OSNR penalty of only 1-dB compared to the single pulse transmission.

  18. Biased allosteric modulation at the CaS receptor engendered by structurally diverse calcimimetics

    PubMed Central

    Cook, A E; Mistry, S N; Gregory, K J; Furness, S G B; Sexton, P M; Scammells, P J; Conigrave, A D; Christopoulos, A; Leach, K

    2015-01-01

    BACKGROUND AND PURPOSE Clinical use of cinacalcet in hyperparathyroidism is complicated by its tendency to induce hypocalcaemia, arising partly from activation of calcium-sensing receptors (CaS receptors) in the thyroid and stimulation of calcitonin release. CaS receptor allosteric modulators that selectively bias signalling towards pathways that mediate desired effects [e.g. parathyroid hormone (PTH) suppression] rather than those mediating undesirable effects (e.g. elevated serum calcitonin), may offer better therapies. EXPERIMENTAL APPROACH We characterized the ligand-biased profile of novel calcimimetics in HEK293 cells stably expressing human CaS receptors, by monitoring intracellular calcium (Ca2+i) mobilization, inositol phosphate (IP)1 accumulation, ERK1/2 phosphorylation (pERK1/2) and receptor expression. KEY RESULTS Phenylalkylamine calcimimetics were biased towards allosteric modulation of Ca2+i mobilization and IP1 accumulation. S,R-calcimimetic B was biased only towards IP1 accumulation. R,R-calcimimetic B and AC-265347 were biased towards IP1 accumulation and pERK1/2. Nor-calcimimetic B was unbiased. In contrast to phenylalkylamines and calcimimetic B analogues, AC-265347 did not promote trafficking of a loss-of-expression, naturally occurring, CaS receptor mutation (G670E). CONCLUSIONS AND IMPLICATIONS The ability of R,R-calcimimetic B and AC-265347 to bias signalling towards pERK1/2 and IP1 accumulation may explain their suppression of PTH levels in vivo at concentrations that have no effect on serum calcitonin levels. The demonstration that AC-265347 promotes CaS receptor receptor signalling, but not trafficking reveals a novel profile of ligand-biased modulation at CaS receptors The identification of allosteric modulators that bias CaS receptor signalling towards distinct intracellular pathways provides an opportunity to develop desirable biased signalling profiles in vivo for mediating selective physiological responses. PMID:25220431

  19. Ignavine: a novel allosteric modulator of the μ opioid receptor

    PubMed Central

    Ohbuchi, Katsuya; Miyagi, Chika; Suzuki, Yasuyuki; Mizuhara, Yasuharu; Mizuno, Keita; Omiya, Yuji; Yamamoto, Masahiro; Warabi, Eiji; Sudo, Yuka; Yokoyama, Akinobu; Miyano, Kanako; Hirokawa, Takatsugu; Uezono, Yasuhito

    2016-01-01

    Processed Aconiti tuber (PAT) is used to treat pain associated with various disorders. Although it has been demonstrated that the κ opioid receptor (KOR) signaling pathway is a mediator of the analgesic effect of PAT, active components affecting opioid signaling have not yet been identified. In this study, we explored candidate components of PAT by pharmacokinetic analysis and identified ignavine, which is a different structure from aconitine alkaloids. A receptor binding assay of opioid receptors showed that ignavine specifically binds the μ opioid receptor (MOR), not the KOR. Receptor internalization assay in MOR-expressing cell lines revealed that ignavine augmented the responses produced by D-Ala(2)-N-Me-Phe(4)-Gly-ol(5)-enkephalin (DAMGO), a representative MOR agonist, at a low concentration and inhibited it at a higher concentration. Ignavine also exerted positive modulatory activity for DAMGO, endomorphin-1 and morphine in cAMP assay. Additionally, ignavine alone showed an analgesic effect in vivo. In silico simulation analysis suggested that ignavine would induce a unique structural change distinguished from those induced by a representative MOR agonist and antagonist. These data collectively suggest the possibility that ignavine could be a novel allosteric modulator of the MOR. The present results may open the way for the development of a novel pain management strategy. PMID:27530869

  20. Are AMPA Receptor Positive Allosteric Modulators Potential Pharmacotherapeutics for Addiction?

    PubMed Central

    Watterson, Lucas R.; Olive, M. Foster

    2013-01-01

    Positive allosteric modulators (PAMs) of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors are a diverse class of compounds that increase fast excitatory transmission in the brain. AMPA PAMs have been shown to facilitate long-term potentiation, strengthen communication between various cortical and subcortical regions, and some of these compounds increase the production and release of brain-derived neurotrophic factor (BDNF) in an activity-dependent manner. Through these mechanisms, AMPA PAMs have shown promise as broad spectrum pharmacotherapeutics in preclinical and clinical studies for various neurodegenerative and psychiatric disorders. In recent years, a small collection of preclinical animal studies has also shown that AMPA PAMs may have potential as pharmacotherapeutic adjuncts to extinction-based or cue-exposure therapies for the treatment of drug addiction. The present paper will review this preclinical literature, discuss novel data collected in our laboratory, and recommend future research directions for the possible development of AMPA PAMs as anti-addiction medications. PMID:24380895

  1. The N-terminal Domain Allosterically Regulates Cleavage and Activation of the Epithelial Sodium Channel*

    PubMed Central

    Kota, Pradeep; Buchner, Ginka; Chakraborty, Hirak; Dang, Yan L.; He, Hong; Garcia, Guilherme J. M.; Kubelka, Jan; Gentzsch, Martina; Stutts, M. Jackson; Dokholyan, Nikolay V.

    2014-01-01

    The epithelial sodium channel (ENaC) is activated upon endoproteolytic cleavage of specific segments in the extracellular domains of the α- and γ-subunits. Cleavage is accomplished by intracellular proteases prior to membrane insertion and by surface-expressed or extracellular soluble proteases once ENaC resides at the cell surface. These cleavage events are partially regulated by intracellular signaling through an unknown allosteric mechanism. Here, using a combination of computational and experimental techniques, we show that the intracellular N terminus of γ-ENaC undergoes secondary structural transitions upon interaction with phosphoinositides. From ab initio folding simulations of the N termini in the presence and absence of phosphatidylinositol 4,5-bisphosphate (PIP2), we found that PIP2 increases α-helical propensity in the N terminus of γ-ENaC. Electrophysiology and mutation experiments revealed that a highly conserved cluster of lysines in the γ-ENaC N terminus regulates accessibility of extracellular cleavage sites in γ-ENaC. We also show that conditions that decrease PIP2 or enhance ubiquitination sharply limit access of the γ-ENaC extracellular domain to proteases. Further, the efficiency of allosteric control of ENaC proteolysis is dependent on Tyr370 in γ-ENaC. Our findings provide an allosteric mechanism for ENaC activation regulated by the N termini and sheds light on a potential general mechanism of channel and receptor activation. PMID:24973914

  2. MCPath: Monte Carlo path generation approach to predict likely allosteric pathways and functional residues

    PubMed Central

    Kaya, Cihan; Armutlulu, Andac; Ekesan, Solen; Haliloglu, Turkan

    2013-01-01

    Allosteric mechanism of proteins is essential in biomolecular signaling. An important aspect underlying this mechanism is the communication pathways connecting functional residues. Here, a Monte Carlo (MC) path generation approach is proposed and implemented to define likely allosteric pathways through generating an ensemble of maximum probability paths. The protein structure is considered as a network of amino acid residues, and inter-residue interactions are described by an atomistic potential function. PDZ domain structures are presented as case studies. The analysis for bovine rhodopsin and three myosin structures are also provided as supplementary case studies. The suggested pathways and the residues constituting the pathways are maximally probable and mostly agree with the previous studies. Overall, it is demonstrated that the communication pathways could be multiple and intrinsically disposed, and the MC path generation approach provides an effective tool for the prediction of key residues that mediate the allosteric communication in an ensemble of pathways and functionally plausible residues. The MCPath server is available at http://safir.prc.boun.edu.tr/clbet_server. PMID:23742907

  3. Emerging Computational Methods for the Rational Discovery of Allosteric Drugs

    PubMed Central

    2016-01-01

    Allosteric drug development holds promise for delivering medicines that are more selective and less toxic than those that target orthosteric sites. To date, the discovery of allosteric binding sites and lead compounds has been mostly serendipitous, achieved through high-throughput screening. Over the past decade, structural data has become more readily available for larger protein systems and more membrane protein classes (e.g., GPCRs and ion channels), which are common allosteric drug targets. In parallel, improved simulation methods now provide better atomistic understanding of the protein dynamics and cooperative motions that are critical to allosteric mechanisms. As a result of these advances, the field of predictive allosteric drug development is now on the cusp of a new era of rational structure-based computational methods. Here, we review algorithms that predict allosteric sites based on sequence data and molecular dynamics simulations, describe tools that assess the druggability of these pockets, and discuss how Markov state models and topology analyses provide insight into the relationship between protein dynamics and allosteric drug binding. In each section, we first provide an overview of the various method classes before describing relevant algorithms and software packages. PMID:27074285

  4. New paradigm for allosteric regulation of Escherichia coli aspartate transcarbamoylase.

    PubMed

    Cockrell, Gregory M; Zheng, Yunan; Guo, Wenyue; Peterson, Alexis W; Truong, Jennifer K; Kantrowitz, Evan R

    2013-11-12

    For nearly 60 years, the ATP activation and the CTP inhibition of Escherichia coli aspartate transcarbamoylase (ATCase) has been the textbook example of allosteric regulation. We present kinetic data and five X-ray structures determined in the absence and presence of a Mg(2+) concentration within the physiological range. In the presence of 2 mM divalent cations (Mg(2+), Ca(2+), Zn(2+)), CTP does not significantly inhibit the enzyme, while the allosteric activation by ATP is enhanced. The data suggest that the actual allosteric inhibitor of ATCase in vivo is the combination of CTP, UTP, and a divalent cation, and the actual allosteric activator is a divalent cation with ATP or ATP and GTP. The structural data reveals that two NTPs can bind to each allosteric site with a divalent cation acting as a bridge between the triphosphates. Thus, the regulation of ATCase is far more complex than previously believed and calls many previous studies into question. The X-ray structures reveal that the catalytic chains undergo essentially no alternations; however, several regions of the regulatory chains undergo significant structural changes. Most significant is that the N-terminal region of the regulatory chains exists in different conformations in the allosterically activated and inhibited forms of the enzyme. Here, a new model of allosteric regulation is proposed.

  5. Allosteric "beta-blocker" isolated from a DNA-encoded small molecule library.

    PubMed

    Ahn, Seungkirl; Kahsai, Alem W; Pani, Biswaranjan; Wang, Qin-Ting; Zhao, Shuai; Wall, Alissa L; Strachan, Ryan T; Staus, Dean P; Wingler, Laura M; Sun, Lillian D; Sinnaeve, Justine; Choi, Minjung; Cho, Ted; Xu, Thomas T; Hansen, Gwenn M; Burnett, Michael B; Lamerdin, Jane E; Bassoni, Daniel L; Gavino, Bryant J; Husemoen, Gitte; Olsen, Eva K; Franch, Thomas; Costanzi, Stefano; Chen, Xin; Lefkowitz, Robert J

    2017-02-14

    The β2-adrenergic receptor (β2AR) has been a model system for understanding regulatory mechanisms of G-protein-coupled receptor (GPCR) actions and plays a significant role in cardiovascular and pulmonary diseases. Because all known β-adrenergic receptor drugs target the orthosteric binding site of the receptor, we set out to isolate allosteric ligands for this receptor by panning DNA-encoded small-molecule libraries comprising 190 million distinct compounds against purified human β2AR. Here, we report the discovery of a small-molecule negative allosteric modulator (antagonist), compound 15 [([4-((2S)-3-(((S)-3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)amino)-2-(2-cyclohexyl-2-phenylacetamido)-3-oxopropyl)benzamide], exhibiting a unique chemotype and low micromolar affinity for the β2AR. Binding of 15 to the receptor cooperatively enhances orthosteric inverse agonist binding while negatively modulating binding of orthosteric agonists. Studies with a specific antibody that binds to an intracellular region of the β2AR suggest that 15 binds in proximity to the G-protein binding site on the cytosolic surface of the β2AR. In cell-signaling studies, 15 inhibits cAMP production through the β2AR, but not that mediated by other Gs-coupled receptors. Compound 15 also similarly inhibits β-arrestin recruitment to the activated β2AR. This study presents an allosteric small-molecule ligand for the β2AR and introduces a broadly applicable method for screening DNA-encoded small-molecule libraries against purified GPCR targets. Importantly, such an approach could facilitate the discovery of GPCR drugs with tailored allosteric effects.

  6. Allosteric “beta-blocker” isolated from a DNA-encoded small molecule library

    PubMed Central

    Ahn, Seungkirl; Kahsai, Alem W.; Pani, Biswaranjan; Wang, Qin-Ting; Zhao, Shuai; Wall, Alissa L.; Strachan, Ryan T.; Staus, Dean P.; Wingler, Laura M.; Sun, Lillian D.; Sinnaeve, Justine; Choi, Minjung; Cho, Ted; Xu, Thomas T.; Hansen, Gwenn M.; Burnett, Michael B.; Lamerdin, Jane E.; Bassoni, Daniel L.; Gavino, Bryant J.; Husemoen, Gitte; Olsen, Eva K.; Franch, Thomas; Costanzi, Stefano; Chen, Xin; Lefkowitz, Robert J.

    2017-01-01

    The β2-adrenergic receptor (β2AR) has been a model system for understanding regulatory mechanisms of G-protein–coupled receptor (GPCR) actions and plays a significant role in cardiovascular and pulmonary diseases. Because all known β-adrenergic receptor drugs target the orthosteric binding site of the receptor, we set out to isolate allosteric ligands for this receptor by panning DNA-encoded small-molecule libraries comprising 190 million distinct compounds against purified human β2AR. Here, we report the discovery of a small-molecule negative allosteric modulator (antagonist), compound 15 [([4-((2S)-3-(((S)-3-(3-bromophenyl)-1-(methylamino)-1-oxopropan-2-yl)amino)-2-(2-cyclohexyl-2-phenylacetamido)-3-oxopropyl)benzamide], exhibiting a unique chemotype and low micromolar affinity for the β2AR. Binding of 15 to the receptor cooperatively enhances orthosteric inverse agonist binding while negatively modulating binding of orthosteric agonists. Studies with a specific antibody that binds to an intracellular region of the β2AR suggest that 15 binds in proximity to the G-protein binding site on the cytosolic surface of the β2AR. In cell-signaling studies, 15 inhibits cAMP production through the β2AR, but not that mediated by other Gs-coupled receptors. Compound 15 also similarly inhibits β-arrestin recruitment to the activated β2AR. This study presents an allosteric small-molecule ligand for the β2AR and introduces a broadly applicable method for screening DNA-encoded small-molecule libraries against purified GPCR targets. Importantly, such an approach could facilitate the discovery of GPCR drugs with tailored allosteric effects. PMID:28130548

  7. Coupled Dynamics and Entropic Contribution to the Allosteric Mechanism of Pin1.

    PubMed

    Barman, Arghya; Hamelberg, Donald

    2016-08-25

    Allosteric communication in proteins regulates a plethora of downstream processes in subcellular signaling pathways. Describing the effects of cooperative ligand binding on the atomic level is a key to understanding many regulatory processes involving biomolecules. Here, we use microsecond-long molecular dynamics simulations to investigate the allosteric mechanism of Pin1, a potential therapeutic target and a phosphorylated-Ser/Thr dependent peptidyl-prolyl cis-trans isomerase that regulates several subcellular processes and has been implicated in many diseases, including cancer and Alzheimer's. Experimental studies suggest that the catalytic domain and the noncatalytic WW domain are allosterically coupled; however, an atomic level description of the dynamics associated with the interdomain communication is lacking. We show that binding of the substrate to the WW domain is directly coupled to the dynamics of the catalytic domain, causing rearrangement of the residue-residue contact dynamics from the WW domain to the catalytic domain. The binding affinity of the substrate in the catalytic domain is also enhanced upon binding of the substrate to the WW domain. Modulation of the dynamics of the catalytic domain upon binding of the substrate to the WW domain leads to prepayment of the entropic cost of binding the substrate to the catalytic domain. This study shows that Ile 28 at the interfacial region between the catalytic and WW domains is certainly one of the residues responsible for bridging the communication between the two domains. The results complement previous experiments and provide valuable atomistic insights into the role of dynamics and possible entropic contribution to the allosteric mechanism of proteins.

  8. Structural dynamics and energetics underlying allosteric inactivation of the cannabinoid receptor CB1

    PubMed Central

    Fay, Jonathan F.; Farrens, David L.

    2015-01-01

    G protein-coupled receptors (GPCRs) are surprisingly flexible molecules that can do much more than simply turn on G proteins. Some even exhibit biased signaling, wherein the same receptor preferentially activates different G-protein or arrestin signaling pathways depending on the type of ligand bound. Why this behavior occurs is still unclear, but it can happen with both traditional ligands and ligands that bind allosterically outside the orthosteric receptor binding pocket. Here, we looked for structural mechanisms underlying these phenomena in the marijuana receptor CB1. Our work focused on the allosteric ligand Org 27569, which has an unusual effect on CB1—it simultaneously increases agonist binding, decreases G-protein activation, and induces biased signaling. Using classical pharmacological binding studies, we find that Org 27569 binds to a unique allosteric site on CB1 and show that it can act alone (without need for agonist cobinding). Through mutagenesis studies, we find that the ability of Org 27569 to bind is related to how much receptor is in an active conformation that can couple with G protein. Using these data, we estimated the energy differences between the inactive and active states. Finally, site-directed fluorescence labeling studies show the CB1 structure stabilized by Org 27569 is different and unique from that stabilized by antagonist or agonist. Specifically, transmembrane helix 6 (TM6) movements associated with G-protein activation are blocked, but at the same time, helix 8/TM7 movements are enhanced, suggesting a possible mechanism for the ability of Org 27569 to induce biased signaling. PMID:26100912

  9. Multi-slice parallel transmission three-dimensional tailored RF (PTX 3DTRF) pulse design for signal recovery in ultra high field functional MRI

    NASA Astrophysics Data System (ADS)

    Zheng, Hai; Zhao, Tiejun; Qian, Yongxian; Schirda, Claudiu; Ibrahim, Tamer S.; Boada, Fernando E.

    2013-03-01

    T2∗ weighted fMRI at high and ultra high field (UHF) is often hampered by susceptibility-induced, through-plane, signal loss. Three-dimensional tailored RF (3DTRF) pulses have been shown to be an effective approach for mitigating through-plane signal loss at UHF. However, the required RF pulse lengths are too long for practical applications. Recently, parallel transmission (PTX) has emerged as a very effective means for shortening the RF pulse duration for 3DTRF without sacrificing the excitation performance. In this article, we demonstrate a RF pulse design strategy for 3DTRF based on the use of multi-slice PTX 3DTRF to simultaneously and precisely recover signal with whole-brain coverage. Phantom and human experiments are used to demonstrate the effectiveness and robustness of the proposed method on three subjects using an eight-channel whole body parallel transmission system.

  10. Ligand Binding to Macromolecules: Allosteric and Sequential Models of Cooperativity.

    ERIC Educational Resources Information Center

    Hess, V. L.; Szabo, Attila

    1979-01-01

    A simple model is described for the binding of ligands to macromolecules. The model is applied to the cooperative binding by hemoglobin and aspartate transcarbamylase. The sequential and allosteric models of cooperative binding are considered. (BB)

  11. Transmission performance of the single-sideband optical OFDM signal with reduced guard-band based on beat interference cancellation receiver

    NASA Astrophysics Data System (ADS)

    Ma, Jianxin; Zheng, Guoli; Zhou, Wei

    2015-12-01

    We have investigated the fiber transmission performance of the single-sideband optical orthogonal frequency division multiplexing (SSB-OOFDM) signal with reduced guard band (GB) received by the beat interference cancellation receiver based on balanced detection (ICRBD). The simulation results show that the error vector magnitude (EVM) of the 40 Gb/s 16-QAM SSB-OOFDM with 2 GHz GB remains below the forward error correction (FEC) limit even after 5×100 km standard single-mode fiber (SSMF) transmission. The link has stable tolerance of both the laser linewidth and the linear channel imperfections.

  12. Inhibition of tumor angiogenesis and growth by a small-molecule multi-FGF receptor blocker with allosteric properties.

    PubMed

    Bono, Françoise; De Smet, Frederik; Herbert, Corentin; De Bock, Katrien; Georgiadou, Maria; Fons, Pierre; Tjwa, Marc; Alcouffe, Chantal; Ny, Annelii; Bianciotto, Marc; Jonckx, Bart; Murakami, Masahiro; Lanahan, Anthony A; Michielsen, Christof; Sibrac, David; Dol-Gleizes, Frédérique; Mazzone, Massimiliano; Zacchigna, Serena; Herault, Jean-Pascal; Fischer, Christian; Rigon, Patrice; Ruiz de Almodovar, Carmen; Claes, Filip; Blanc, Isabelle; Poesen, Koen; Zhang, Jie; Segura, Inmaculada; Gueguen, Geneviève; Bordes, Marie-Françoise; Lambrechts, Diether; Broussy, Roselyne; van de Wouwer, Marlies; Michaux, Corinne; Shimada, Toru; Jean, Isabelle; Blacher, Silvia; Noel, Agnès; Motte, Patrick; Rom, Eran; Rakic, Jean-Marie; Katsuma, Susumu; Schaeffer, Paul; Yayon, Avner; Van Schepdael, Ann; Schwalbe, Harald; Gervasio, Francesco Luigi; Carmeliet, Geert; Rozensky, Jef; Dewerchin, Mieke; Simons, Michael; Christopoulos, Arthur; Herbert, Jean-Marc; Carmeliet, Peter

    2013-04-15

    Receptor tyrosine kinases (RTK) are targets for anticancer drug development. To date, only RTK inhibitors that block orthosteric binding of ligands and substrates have been developed. Here, we report the pharmacologic characterization of the chemical SSR128129E (SSR), which inhibits fibroblast growth factor receptor (FGFR) signaling by binding to the extracellular FGFR domain without affecting orthosteric FGF binding. SSR exhibits allosteric properties, including probe dependence, signaling bias, and ceiling effects. Inhibition by SSR is highly conserved throughout the animal kingdom. Oral delivery of SSR inhibits arthritis and tumors that are relatively refractory to anti-vascular endothelial growth factor receptor-2 antibodies. Thus, orally-active extracellularly acting small-molecule modulators of RTKs with allosteric properties can be developed and may offer opportunities to improve anticancer treatment.

  13. Fading-free transmission of 124-Gb/s PDM-DMT signal over 100-km SSMF using digital carrier regeneration.

    PubMed

    Li, Cai; Hu, Rong; Yang, Qi; Luo, Ming; Li, Wei; Yu, Shaohua

    2016-01-25

    The coherent reception of intensity modulated signal has been recently widely investigated, in which the signal is recovered by the envelop detection. High linewidth tolerance is achieved with such scheme. However, strong optical carrier exists during the transmission, which degrades the optical power efficiency. In this paper, an efficient modulation scheme for discrete multi-tone (DMT) signal is proposed based on the Mach-Zehnder modulator (MZM). Different from the traditional intensity modulation, the proposed method employs both intensity and phase domain. Thus, the optical carrier power can be greatly reduced by adjusting the bias of MZM around the null point. By employing coherent detection and digital carrier regeneration (DCR), the carrier suppressed DMT signal can be recovered using envelop detection. No carrier frequency or phase estimation is required. Numerical investigations are made to demonstrate the feasibility, in which significant improvements are found for the proposed DCR method, showing great tolerance against laser linewidth and carrier power reduction. Finally, a 124-Gb/s transmission of polarization-division multiplexed DMT (PDM-DMT) signal is demonstrated over 100-km SSMF, with only -8 dB optical carrier to signal power ratio (CSPR).

  14. Transmission of electric and magnetic foetal cardiac signals in a case of ectopia cordis: the dominant role of the vernix. caseosa.

    PubMed

    Wakai, R T; Lengle, J M; Leuthold, A C

    2000-07-01

    Foetal electrocardiograms (fECGs) and foetal magnetocardiograms (fMCGs) were recorded in the 26th, 29th and 31st weeks of gestation from a foetus with ectopia cordis-a rare condition in which the heart lies outside the chest wall. This provided an opportunity to study foetal cardiograms uninfluenced by the insulating effects of the foetal skin and vernix caseosa. The fECG of the ectopia cordis foetus was striking. Unlike recordings from age-matched normal foetuses, recordings from this subject had very high signal-to-noise ratio and showed no anomalous signal transmission properties. In contrast, fMCGs recorded from the ectopia cordis foetus and normal foetuses were largely similar. Both showed high signal-to-noise ratio and signal transmission properties consistent with volume conduction. The findings corroborate the hypothesis that high foetal skin resistance due primarily to the vernix caseosa is responsible for the low amplitude and anomalous transmission properties of the normal fECG, and demonstrate that the fMCG is relatively insensitive to conductivity inhomogeneities.

  15. Design consideration of a 77-channel 64-QAM signal transmission system using a light-injected MQW-SOA as an in-line amplifier

    NASA Astrophysics Data System (ADS)

    Tzeng, SohnLing; Tai, Chien; Chang, Hung-Chun

    1998-06-01

    In the future digital video transmission, a multichannel subcarrier multiplexed system with M-ary quadrature- amplitude-modulation (M-QAM) signals over fiber-optic transmission is an excellent candidate. A system using 1.3 micrometers semiconductor optical amplifiers (SOAs) for increasing the span length or for power splitting is very economical at the situations of the penetration of 1.3-micrometers single-mode fiber with the having installed 1.3-micrometers optical transmitters. However, the significant intermodulation distortions (IMDs) resulted from the signal-induced carrier density modulation of SOA limits the system capability, including channel capacity and power budget. A method using the external light injection to the SOA used in a 77-channel 64-QAM signal transmission system as an in-line amplifier is capable of increasing system power budget. From the viewpoint of system design, two approaches are addressed, one is the optimization of the in-line SOA used the external light injection, the other is the consideration of the optical modulation index per channel (OMI/ch) applied to transmitted. From the experimental results, the pumping wavelength and the pump power in approach one are mainly decided by the requirement of BER and the dynamic range of SOA, in addition, choosing the pump resulting in a smaller accompanied signal gain reduction can enlarge the system power budget. In approach, the optimum position of the in- line SOA incorporated is determined by the value of applied OMI/ch.

  16. Positive Allosteric Modulators of Metabotropic Glutamate 2 Receptors in Schizophrenia Treatment

    PubMed Central

    Ellaithy, Amr; Younkin, Jason; Gonzalez-Maeso, Javier; Logothetis, Diomedes E.

    2015-01-01

    The last two decades have witnessed a rise in the “NMDA receptor hypofunction” hypothesis for schizophrenia, a devastating disorder that affects around 1% of the population worldwide. A variety of presynaptic, postsynaptic and regulatory proteins involved in glutamatergic signaling have thus been proposed as potential therapeutic targets. This Review focuses on positive allosteric modulation of metabotropic glutamate 2 receptors (mGlu2Rs) and discusses how recent preclinical epigenetic data may provide a molecular explanation for the discrepant results of clinical studies, further stimulating the field to exploit the promise of mGlu2R as a target for schizophrenia treatment. PMID:26148747

  17. Discovery of Novel Allosteric Effectors Based on the Predicted Allosteric Sites for Escherichia coli D-3-Phosphoglycerate Dehydrogenase

    PubMed Central

    Wang, Qian; Qi, Yifei; Yin, Ning; Lai, Luhua

    2014-01-01

    D-3-phosphoglycerate dehydrogenase (PGDH) from Escherichia coli catalyzes the first critical step in serine biosynthesis, and can be allosterically inhibited by serine. In a previous study, we developed a computational method for allosteric site prediction using a coarse-grained two-state Gō Model and perturbation. Two potential allosteric sites were predicted for E. coli PGDH, one close to the active site and the nucleotide binding site (Site I) and the other near the regulatory domain (Site II). In the present study, we discovered allosteric inhibitors and activators based on site I, using a high-throughput virtual screen, and followed by using surface plasmon resonance (SPR) to eliminate false positives. Compounds 1 and 2 demonstrated a low-concentration activation and high-concentration inhibition phenomenon, with IC50 values of 34.8 and 58.0 µM in enzymatic bioassays, respectively, comparable to that of the endogenous allosteric effector, L-serine. For its activation activity, compound 2 exhibited an AC50 value of 34.7 nM. The novel allosteric site discovered in PGDH was L-serine- and substrate-independent. Enzyme kinetics studies showed that these compounds influenced Km, kcat, and kcat/Km. We have also performed structure-activity relationship studies to discover high potency allosteric effectors. Compound 2-2, an analog of compound 2, showed the best in vitro activity with an IC50 of 22.3 µM. Compounds targeting this site can be used as new chemical probes to study metabolic regulation in E. coli. Our study not only identified a novel allosteric site and effectors for PGDH, but also provided a general strategy for designing new regulators for metabolic enzymes. PMID:24733054

  18. Ryanodine Receptor Allosteric Coupling and the Dynamics of Calcium Sparks

    PubMed Central

    Groff, Jeffrey R.; Smith, Gregory D.

    2008-01-01

    Puffs and sparks are localized intracellular Ca2+ elevations that arise from the cooperative activity of Ca2+-regulated inositol 1,4,5-trisphosphate receptors and ryanodine receptors clustered at Ca2+ release sites on the surface of the endoplasmic reticulum or the sarcoplasmic reticulum. While the synchronous gating of Ca2+-regulated Ca2+ channels can be mediated entirely though the buffered diffusion of intracellular Ca2+, interprotein allosteric interactions also contribute to the dynamics of ryanodine receptor (RyR) gating and Ca2+ sparks. In this article, Markov chain models of Ca2+ release sites are used to investigate how the statistics of Ca2+ spark generation and termination are related to the coupling of RyRs via local [Ca2+] changes and allosteric interactions. Allosteric interactions are included in a manner that promotes the synchronous gating of channels by stabilizing neighboring closed-closed and/or open-open channel pairs. When the strength of Ca2+-mediated channel coupling is systematically varied (e.g., by changing the Ca2+ buffer concentration), simulations that include synchronizing allosteric interactions often exhibit more robust Ca2+ sparks; however, for some Ca2+ coupling strengths the sparks are less robust. We find no evidence that the distribution of spark durations can be used to distinguish between allosteric interactions that stabilize closed channel pairs, open channel pairs, or both in a balanced fashion. On the other hand, the changes in spark duration, interspark interval, and frequency observed when allosteric interactions that stabilize closed channel pairs are gradually removed from simulations are qualitatively different than the changes observed when open or both closed and open channel pairs are stabilized. Thus, our simulations clarify how changes in spark statistics due to pharmacological washout of the accessory proteins mediating allosteric coupling may indicate the type of synchronizing allosteric interactions exhibited

  19. Rafoxanide and Closantel Inhibit SPAK and OSR1 Kinases by Binding to a Highly Conserved Allosteric Site on Their C-terminal Domains.

    PubMed

    AlAmri, Mubarak A; Kadri, Hachemi; Alderwick, Luke J; Simpkins, Nigel S; Mehellou, Youcef

    2017-03-31

    SPAK and OSR1 are two protein kinases that have emerged as attractive targets in the discovery of novel antihypertensive agents due to their role in regulating electrolyte balance in vivo. Herein we report the identification of an allosteric pocket on the highly conserved C-terminal domains of these two kinases, which influences their activity. We also show that some known WNK signaling inhibitors bind to this allosteric site. Using in silico screening, we identified the antiparasitic agent rafoxanide as a novel allosteric inhibitor of SPAK and OSR1. Collectively, this work will facilitate the rational design of novel SPAK and OSR1 kinase inhibitors that could be useful antihypertensive agents.

  20. Allosteric Voltage Gating of Potassium Channels I

    PubMed Central

    Horrigan, Frank T.; Cui, Jianmin; Aldrich, Richard W.

    1999-01-01

    Activation of large conductance Ca2+-activated K+ channels is controlled by both cytoplasmic Ca2+ and membrane potential. To study the mechanism of voltage-dependent gating, we examined mSlo Ca2+-activated K+ currents in excised macropatches from Xenopus oocytes in the virtual absence of Ca2+ (<1 nM). In response to a voltage step, IK activates with an exponential time course, following a brief delay. The delay suggests that rapid transitions precede channel opening. The later exponential time course suggests that activation also involves a slower rate-limiting step. However, the time constant of IK relaxation [τ(IK)] exhibits a complex voltage dependence that is inconsistent with models that contain a single rate limiting step. τ(IK) increases weakly with voltage from −500 to −20 mV, with an equivalent charge (z) of only 0.14 e, and displays a stronger voltage dependence from +30 to +140 mV (z = 0.49 e), which then decreases from +180 to +240 mV (z = −0.29 e). Similarly, the steady state GK–V relationship exhibits a maximum voltage dependence (z = 2 e) from 0 to +100 mV, and is weakly voltage dependent (z ≅ 0.4 e) at more negative voltages, where Po = 10−5–10−6. These results can be understood in terms of a gating scheme where a central transition between a closed and an open conformation is allosterically regulated by the state of four independent and identical voltage sensors. In the absence of Ca2+, this allosteric mechanism results in a gating scheme with five closed (C) and five open (O) states, where the majority of the channel's voltage dependence results from rapid C–C and O–O transitions, whereas the C–O transitions are rate limiting and weakly voltage dependent. These conclusions not only provide a framework for interpreting studies of large conductance Ca2+-activated K+ channel voltage gating, but also have important implications for understanding the mechanism of Ca2+ sensitivity. PMID:10436003

  1. Demonstration of 48-Gb/s 16-QAM signal transmission using half cycle sub-carrier modulation in intensity modulation/direct detection system

    NASA Astrophysics Data System (ADS)

    Tang, Jin; He, Jing; Chen, Ming; Li, Danyu; Chen, Lin

    2015-01-01

    A simple spectral-efficiency intensity modulation/direct detection (IM/DD) system based on half cycle sub-carrier modulation (SCM) signal is proposed for short reach fiber communications in this paper. The signal impairment of frequency selective fading due to fiber chromatics dispersion (CD) is mathematically analyzed. To reduce the performance deterioration caused by the non-flat transfer function, digital pre- and post-equalization is applied in the system. The peak to average power ratio (PAPR) of the signal is also discussed in comparison with that of orthogonal frequency division multiplexing (OFDM). The transmission of 16-QAM half cycle SCM signal with a sub-carrier frequency of half the symbol rate and Nyquist pulse shaping is experimentally demonstrated. The bit-error rate (BER) of 48 Gb/s polarization multiplexing division (PDM) 16 QAM half cycle SCM signal is less than 7% forward-error-correction (FEC) threshold of 3.8 ×10-3 after transmission over 83 km standard single-mode fiber (SSMF).

  2. Allosteric regulation of rhomboid intramembrane proteolysis.

    PubMed

    Arutyunova, Elena; Panwar, Pankaj; Skiba, Pauline M; Gale, Nicola; Mak, Michelle W; Lemieux, M Joanne

    2014-09-01

    Proteolysis within the lipid bilayer is poorly understood, in particular the regulation of substrate cleavage. Rhomboids are a family of ubiquitous intramembrane serine proteases that harbour a buried active site and are known to cleave transmembrane substrates with broad specificity. In vitro gel and Förster resonance energy transfer (FRET)-based kinetic assays were developed to analyse cleavage of the transmembrane substrate psTatA (TatA from Providencia stuartii). We demonstrate significant differences in catalytic efficiency (kcat/K0.5) values for transmembrane substrate psTatA (TatA from Providencia stuartii) cleavage for three rhomboids: AarA from P. stuartii, ecGlpG from Escherichia coli and hiGlpG from Haemophilus influenzae demonstrating that rhomboids specifically recognize this substrate. Furthermore, binding of psTatA occurs with positive cooperativity. Competitive binding studies reveal an exosite-mediated mode of substrate binding, indicating allostery plays a role in substrate catalysis. We reveal that exosite formation is dependent on the oligomeric state of rhomboids, and when dimers are dissociated, allosteric substrate activation is not observed. We present a novel mechanism for specific substrate cleavage involving several dynamic processes including positive cooperativity and homotropic allostery for this interesting class of intramembrane proteases.

  3. Allosteric pathway identification through network analysis: from molecular dynamics simulations to interactive 2D and 3D graphs.

    PubMed

    Allain, Ariane; Chauvot de Beauchêne, Isaure; Langenfeld, Florent; Guarracino, Yann; Laine, Elodie; Tchertanov, Luba

    2014-01-01

    Allostery is a universal phenomenon that couples the information induced by a local perturbation (effector) in a protein to spatially distant regulated sites. Such an event can be described in terms of a large scale transmission of information (communication) through a dynamic coupling between structurally rigid (minimally frustrated) and plastic (locally frustrated) clusters of residues. To elaborate a rational description of allosteric coupling, we propose an original approach - MOdular NETwork Analysis (MONETA) - based on the analysis of inter-residue dynamical correlations to localize the propagation of both structural and dynamical effects of a perturbation throughout a protein structure. MONETA uses inter-residue cross-correlations and commute times computed from molecular dynamics simulations and a topological description of a protein to build a modular network representation composed of clusters of residues (dynamic segments) linked together by chains of residues (communication pathways). MONETA provides a brand new direct and simple visualization of protein allosteric communication. A GEPHI module implemented in the MONETA package allows the generation of 2D graphs of the communication network. An interactive PyMOL plugin permits drawing of the communication pathways between chosen protein fragments or residues on a 3D representation. MONETA is a powerful tool for on-the-fly display of communication networks in proteins. We applied MONETA for the analysis of communication pathways (i) between the main regulatory fragments of receptors tyrosine kinases (RTKs), KIT and CSF-1R, in the native and mutated states and (ii) in proteins STAT5 (STAT5a and STAT5b) in the phosphorylated and the unphosphorylated forms. The description of the physical support for allosteric coupling by MONETA allowed a comparison of the mechanisms of (a) constitutive activation induced by equivalent mutations in two RTKs and (b) allosteric regulation in the activated and non

  4. Colorless WDM-PON based on a Fabry-Pérot laser diode and reflective semiconductor optical amplifiers for simultaneous transmission of bidirectional gigabit baseband signals and broadcasting signal.

    PubMed

    Pham, Thang Tien; Kim, Hyun-Seung; Won, Yong-Yuk; Han, Sang-Kook

    2009-09-14

    A novel WDM-PON system delivering bidirectional baseband data and broadcasting data is proposed and demonstrated. A subcarrier multiplexing signal is broadcasted to all users by modulating a broadband optical source based on a Fabry-Pérot laser diode. Reflective semiconductor optical amplifiers are used as colorless modulators for the baseband data at both optical line terminal and remote optical network units. Transmission performance including bit error rate of bidirectional gigabit data and error vector magnitude of broadcasting data of many optical channels is investigated. Additionally, the data rate for the broadcasting signal was improved by using an external modulator.

  5. Transmission of 112(4×28)-Gb/s PAM-4 Signal over 48.6-km SSMF within only 50-GHz Grid

    NASA Astrophysics Data System (ADS)

    Dong, Jian; Hu, Rong

    2016-12-01

    In this paper, a transmission of 112(4×28)-Gb/s PAM-4 signal is experimentally demonstrated within only 50-GHz grid, achieving an optical spectral efficiency (SE) of 2.24 b/s/Hz. For the intensity modulation and direct detection (IM/DD) based PAM-4 transmission, it is the first time to the best of our knowledge that 112-Gb/s PAM-4 signal has been transmitted over 48.6-km standard single mode fiber (SSMF), which is compatible with 50-GHz standard gridding. By employing digital pre-equalization, duobinary encoding/decoding and 7-level based training sequence aided least-mean square (TS-LMS) algorithm, each lane of the 28-Gb/s PAM-4 signal occupies less than 11-GHz optical spectral (3-dB bandwidth), resulting in negligible inter-channel interference for the 4-lane 112-Gb/s PAM-4 signals within 50-GHz grid. The proposed method is bandwidth and computationally efficient, which is thought feasible in the low-cost short reach optical networks.

  6. Activation of nanoscale allosteric protein domain motion revealed by neutron spin echo spectroscopy

    NASA Astrophysics Data System (ADS)

    Bu, Zimei; Farago, Bela; Callaway, David

    2012-02-01

    NHERF1 is a multi-domain scaffolding protein that assembles the signaling complexes, and regulates the cell surface expression and endocytic recycling of a variety of membrane proteins. The ability of the two PDZ domains in NHERF1 to assemble protein complexes is allosterically modulated by a membrane-cytoskeleton linker protein ezrin, whose binding site is located as far as 110 angstroms away from the PDZ domains. Here, using neutron spin echo (NSE) spectroscopy, selective deuterium labeling, and theoretical analyses, we reveal the activation of interdomain motion in NHERF1 on nanometer length scales and on sub-microsecond time scales upon forming a complex with ezrin. We show that a much simplified coarse-grained model is sufficient to describe inter-domain motion of a multi-domain protein or protein complex. We expect that future NSE experiments will benefit by exploiting our approach of selective deuteration to resolve the specific domain motions of interest from a plethora of global translational and rotational motions. The results demonstrate that propagation of allosteric signals to distal sites involves the activation of long-range coupled domain motions on submicrosecond time scales, and that these coupled motions can be distinguished and characterized by NSE.

  7. Continuous Allosteric Regulation of a Viral Packaging Motor by a Sensor that Detects the Density and Conformation of Packaged DNA

    PubMed Central

    Berndsen, Zachary T.; Keller, Nicholas; Smith, Douglas E.

    2015-01-01

    We report evidence for an unconventional type of allosteric regulation of a biomotor. We show that the genome-packaging motor of phage ϕ29 is regulated by a sensor that detects the density and conformation of the DNA packaged inside the viral capsid, and slows the motor by a mechanism distinct from the effect of a direct load force on the motor. Specifically, we show that motor-ATP interactions are regulated by a signal that is propagated allosterically from inside the viral shell to the motor mounted on the outside. This signal continuously regulates the motor speed and pausing in response to changes in either density or conformation of the packaged DNA, and slows the motor before the buildup of large forces resisting DNA confinement. Analysis of motor slipping reveals that the force resisting packaging remains low (<1 pN) until ∼70% and then rises sharply to ∼23 pN at high filling, which is a several-fold lower value than was previously estimated under the assumption that force alone slows the motor. These findings are consistent with recent studies of the stepping kinetics of the motor. The allosteric regulatory mechanism we report allows double-stranded DNA viruses to achieve rapid, high-density packing of their genomes by limiting the buildup of nonequilibrium load forces on the motor. PMID:25606680

  8. Seven Transmembrane Receptors as Shapeshifting Proteins: The Impact of Allosteric Modulation and Functional Selectivity on New Drug Discovery

    PubMed Central

    Miller, Laurence J.

    2010-01-01

    It is useful to consider seven transmembrane receptors (7TMRs) as disordered proteins able to allosterically respond to a number of binding partners. Considering 7TMRs as allosteric systems, affinity and efficacy can be thought of in terms of energy flow between a modulator, conduit (the receptor protein), and a number of guests. These guests can be other molecules, receptors, membrane-bound proteins, or signaling proteins in the cytosol. These vectorial flows of energy can yield standard canonical guest allostery (allosteric modification of drug effect), effects along the plane of the cell membrane (receptor oligomerization), or effects directed into the cytosol (differential signaling as functional selectivity). This review discusses these apparently diverse pharmacological effects in terms of molecular dynamics and protein ensemble theory, which tends to unify 7TMR behavior toward cells. Special consideration will be given to functional selectivity (biased agonism and biased antagonism) in terms of mechanism of action and potential therapeutic application. The explosion of technology that has enabled observation of diverse 7TMR behavior has also shown how drugs can have multiple (pluridimensional) efficacies and how this can cause paradoxical drug classification and nomenclatures. PMID:20392808

  9. Structure, Dynamics, and Allosteric Potential of Ionotropic Glutamate Receptor N-Terminal Domains

    PubMed Central

    Krieger, James; Bahar, Ivet; Greger, Ingo H.

    2015-01-01

    Ionotropic glutamate receptors (iGluRs) are tetrameric cation channels that mediate synaptic transmission and plasticity. They have a unique modular architecture with four domains: the intracellular C-terminal domain (CTD) that is involved in synaptic targeting, the transmembrane domain (TMD) that forms the ion channel, the membrane-proximal ligand-binding domain (LBD) that binds agonists such as L-glutamate, and the distal N-terminal domain (NTD), whose function is the least clear. The extracellular portion, comprised of the LBD and NTD, is loosely arranged, mediating complex allosteric regulation and providing a rich target for drug development. Here, we briefly review recent work on iGluR NTD structure and dynamics, and further explore the allosteric potential for the NTD in AMPA-type iGluRs using coarse-grained simulations. We also investigate mechanisms underlying the established NTD allostery in NMDA-type iGluRs, as well as the fold-related metabotropic glutamate and GABAB receptors. We show that the clamshell motions intrinsically favored by the NTD bilobate fold are coupled to dimeric and higher-order rearrangements that impact the iGluR LBD and ultimately the TMD. Finally, we explore the dynamics of intact iGluRs and describe how it might affect receptor operation in a synaptic environment. PMID:26255587

  10. An allosteric inhibitor of substrate recognition by the SCF[superscript Cdc4] ubiquitin ligase

    SciTech Connect

    Orlicky, Stephen; Tang, Xiaojing; Neduva, Victor; Elowe, Nadine; Brown, Eric D.; Sicheri, Frank; Tyers, Mike

    2010-09-17

    The specificity of SCF ubiquitin ligase-mediated protein degradation is determined by F-box proteins. We identified a biplanar dicarboxylic acid compound, called SCF-I2, as an inhibitor of substrate recognition by the yeast F-box protein Cdc4 using a fluorescence polarization screen to monitor the displacement of a fluorescein-labeled phosphodegron peptide. SCF-I2 inhibits the binding and ubiquitination of full-length phosphorylated substrates by SCF{sup Cdc4}. A co-crystal structure reveals that SCF-I2 inserts itself between the {beta}-strands of blades 5 and 6 of the WD40 propeller domain of Cdc4 at a site that is 25 {angstrom} away from the substrate binding site. Long-range transmission of SCF-I2 interactions distorts the substrate binding pocket and impedes recognition of key determinants in the Cdc4 phosphodegron. Mutation of the SCF-I2 binding site abrogates its inhibitory effect and explains specificity in the allosteric inhibition mechanism. Mammalian WD40 domain proteins may exhibit similar allosteric responsiveness and hence represent an extensive class of druggable target.

  11. 400Gb/s (4 x 100Gb/s) orthogonal PDM-RZ-QPSK DWDM signal transmission over 1040km SMF-28.

    PubMed

    Yu, Jianjun; Zhou, Xiang; Huang, Ming-Fang; Qian, Dayou; Ji, Philip N; Wang, Ting; Magill, Peter

    2009-09-28

    We have generated 4 x 100-Gb/s orthogonal WDM optical signal by employing polarization-division-multiplexed (PDM) return-to-zero (RZ) QPSK modulation format and tight optical filtering technique. The required optical signal-to-noise ratio (OSNR) at bit error ratio (BER) of 2 x 10(-3) for the 400 Gb/s orthogonal DWDM signal is measured to be approximately 22.8 dB/0.1 nm. After transmission over 1040-km standard single mode fiber (EDFA-only amplification, 80-km amplifier span and fully receiver-side electrical dispersion compensation), the measured BER for all the four orthogonal subchannels are smaller than 2 x 10(-3).

  12. Kerr nonlinearity mitigation in 5 × 28-GBd PDM 16-QAM signal transmission over a dispersion-uncompensated link with backward-pumped distributed Raman amplification.

    PubMed

    Sackey, Isaac; Da Ros, Francesco; Jazayerifar, Mahmoud; Richter, Thomas; Meuer, Christian; Nölle, Markus; Molle, Lutz; Peucheret, Christophe; Petermann, Klaus; Schubert, Colja

    2014-11-03

    We present experimental and numerical investigations of Kerr nonlinearity compensation in a 400-km standard single-mode fiber link with distributed Raman amplification with backward pumping. A dual-pump polarization-independent fiber-based optical parametric amplifier is used for mid-link spectral inversion of 5 × 28-GBd polarization-multiplexed 16-QAM signals. Signal quality factor (Q-factor) improvements of 1.1 dB and 0.8 dB were obtained in the cases of a single-channel and a five-channel wavelength-division multiplexing (WDM) system, respectively. The experimental results are compared to numerical simulations with good agreement. It is also shown with simulations that a maximum transmission reach of 2400 km enabled by the optical phase conjugator is possible for the WDM signal.

  13. Evidence of the Role of R-Spondin 1 and Its Receptor Lgr4 in the Transmission of Mechanical Stimuli to Biological Signals for Bone Formation

    PubMed Central

    Shi, Gui-Xun; Zheng, Xin-Feng; Zhu, Chao; Li, Bo; Wang, Yu-Ren; Jiang, Sheng-Dan; Jiang, Lei-Sheng

    2017-01-01

    The bone can adjust its mass and architecture to mechanical stimuli via a series of molecular cascades, which have been not yet fully elucidated. Emerging evidence indicated that R-spondins (Rspos), a family of secreted agonists of the Wnt/β-catenin signaling pathway, had important roles in osteoblastic differentiation and bone formation. However, the role of Rspo proteins in mechanical loading-influenced bone metabolism has never been investigated. In this study, we found that Rspo1 was a mechanosensitive protein for bone formation. Continuous cyclic mechanical stretch (CMS) upregulated the expression of Rspo1 in mouse bone marrow mesenchymal stem cells (BMSCs), while the expression of Rspo1 in BMSCs in vivo was downregulated in the bones of a mechanical unloading mouse model (tail suspension (TS)). On the other hand, Rspo1 could promote osteogenesis of BMSCs under CMS through activating the Wnt/β-catenin signaling pathway and could rescue the bone loss induced by mechanical unloading in the TS mice. Specifically, our results suggested that Rspo1 and its receptor of leucine-rich repeat containing G-protein-coupled receptor 4 (Lgr4) should be a novel molecular signal in the transmission of mechanical stimuli to biological signal in the bone, and this signal should be in the upstream of Wnt/β-catenin signaling for bone formation. Rspo1/Lgr4 could be a new potential target for the prevention and treatment of disuse osteoporosis in the future. PMID:28272338

  14. Identification of an allosteric modulator of the serotonin transporter with novel mechanism of action.

    PubMed

    Kortagere, Sandhya; Fontana, Andreia Cristina Karklin; Rose, Deja Renée; Mortensen, Ole Valente

    2013-09-01

    Serotonin transporters (SERTs) play an essential role in the termination and regulation of serotonin signaling in the brain. SERT is also the target of antidepressants and psychostimulants. Molecules with novel activities and modes of interaction with regard to SERT function are of great scientific and clinical interest. We explored structural regions outside the putative serotonin translocation pathway to identify potential binding sites for allosteric transporter modulators (ATMs). Mutational studies revealed a pocket of amino acids outside the orthosteric substrate binding sites located in the interface between extracellular loops 1 and 3 that when mutated affect transporter function. Using the structure of the bacterial transporter homolog leucine transporter as a template, we developed a structural model of SERT. We performed molecular dynamics simulations to further characterize the allosteric pocket that was identified by site-directed mutagenesis studies and employed this pocket in a virtual screen for small-molecule modulators of SERT function. In functional transport assays, we found that one of the identified molecules, ATM7, increased the reuptake of serotonin, possibly by facilitating the interaction of serotonin with transport-ready conformations of SERT when concentrations of serotonin were low and rate limiting. In addition, ATM7 potentiates 3,4-methylenedioxy-N-methylamphetamine (MDMA, "Ecstasy")-induced reversed transport by SERT. Taking advantage of a conformationally sensitive residue in transmembrane domain 6, we demonstrate that ATM7 mechanistically stabilizes an outward-facing conformation of SERT. Taken together these observations demonstrate that ATM7 acts through a novel mechanism that involves allosteric modulation of SERT function.

  15. Allosteric dynamics of SAMHD1 studied by molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Patra, K. K.; Bhattacharya, A.; Bhattacharya, S.

    2016-10-01

    SAMHD1 is a human cellular enzyme that blocks HIV-1 infection in myeloid cells and non-cycling CD4+T cells. The enzyme is an allosterically regulated triphosphohydrolase that modulates the level of cellular dNTP. The virus restriction is attributed to the lowering of the pool of dNTP in the cell to a point where reverse-transcription is impaired. Mutations in SAMHD1 are also implicated in Aicardi-Goutieres syndrome. A mechanistic understanding of the allosteric activation of the enzyme is still elusive. We have performed molecular dynamics simulations to examine the allosteric site dynamics of the protein and to examine the connection between the stability of the tetrameric complex and the Allosite occupancy.

  16. Experimental demonstration of 608Gbit/s short reach transmission employing half-cycle 16QAM Nyquist-SCM signal and direct detection with 25Gbps EML.

    PubMed

    Zhong, Kangping; Zhou, Xian; Wang, Yiguang; Wang, Liang; Yuan, Jinhui; Yu, Changyuan; Lau, Alan Pak Tao; Lu, Chao

    2016-10-31

    In this paper, we experimentally demonstrated an IM/DD short reach transmission system with a total capacity of 608Gbit/s (net capacity of 565.4Gbit/s exclude 7% FEC overhead) employing half-cycle 16QAM Nyquist-SCM signal and 25Gbps EML at O band. Direct detection-faster than Nyquist (DD-FTN) technique was employed to compensate channel impairments. Number of taps of DD-LMS and tap coefficient of post filter in DD-FTN were experimentally studied for different baud rates. Single-lane 152Gbit/s transmission over 10km of SSMF was experimentally demonstrated. Employing a 4-lanes LAN-WDM architecture, a total capacity of 608Gbit/s transmission over 2km was successfully achieved with a receiver sensitivity lower than -4dBm. To the best of authors' knowledge, this is the highest reported baud rate of half-cycle 16QAM Nyquist-SCM signal and the highest bit rate employing IM/DD and 25Gbps EML in a four lanes LAN-WDM architecture for short reach systems in the O band.

  17. RNA-mediated gene silencing signals are not graft transmissible from the rootstock to the scion in greenhouse-grown apple plants Malus sp.

    PubMed

    Flachowsky, Henryk; Tränkner, Conny; Szankowski, Iris; Waidmann, Sascha; Hanke, Magda-Viola; Treutter, Dieter; Fischer, Thilo C

    2012-01-01

    RNA silencing describes the sequence specific degradation of RNA targets. Silencing is a non-cell autonomous event that is graft transmissible in different plant species. The present study is the first report on systemic acquired dsRNA-mediated gene silencing of transgenic and endogenous gene sequences in a woody plant like apple. Transgenic apple plants overexpressing a hairpin gene construct of the gusA reporter gene were produced. These plants were used as rootstocks and grafted with scions of the gusA overexpressing transgenic apple clone T355. After grafting, we observed a reduction of the gusA gene expression in T355 scions in vitro, but not in T355 scions grown in the greenhouse. Similar results were obtained after silencing of the endogenous Mdans gene in apple that is responsible for anthocyanin biosynthesis. Subsequently, we performed grafting experiments with Mdans silenced rootstocks and red leaf scions of TNR31-35 in order to evaluate graft transmitted silencing of the endogenous Mdans. The results obtained suggested a graft transmission of silencing signals in in vitro shoots. In contrast, no graft transmission of dsRNA-mediated gene silencing signals was detectable in greenhouse-grown plants and in plants grown in an insect protection tent.

  18. Space-division-multiplexed transmission of 3x3 multiple-input multiple-output wireless signals over conventional graded-index multimode fiber.

    PubMed

    Lei, Yi; Li, Jianqiang; Fan, Yuting; Yu, Dawei; Fu, Songnian; Yin, Feifei; Dai, Yitang; Xu, Kun

    2016-12-12

    In this paper, we experimentally demonstrate space-division-multiplexed (SDM) transmission of IEEE 802.11ac-compliant 3-spatial-stream WLAN signals over 3 spatial modes of conventional 50um graded-index (GI) multimode fiber (MMF) employing non-mode-selective 3D-waveguide photonic lantern. Two kinds of scenarios, including fiber-only transmission and fiber-wireless hybrid transmission, were investigated by measuring error vector magnitude (EVM) performance for each stream and condition number (CN) of the channel matrix. The experimental results show that, SDM-based MMF link could offer a CN< 20dB well-conditioned MIMO channel over up to 1km fiber length within 0-6GHz, achieving as low as 2.38%, 2.97% and 2.11% EVM performance for 1km MMF link at 2.4GHz, 5.8GHz, and 200m MMF link followed by 1m air distance at 2.7GHz, respectively. These results indicate the possibility to distribute wireless MIMO signals over existing in-building commercially-available MMFs with enormous cost-saving.

  19. A multi-channel low-power system-on-chip for single-unit recording and narrowband wireless transmission of neural signal.

    PubMed

    Bonfanti, A; Ceravolo, M; Zambra, G; Gusmeroli, R; Spinelli, A S; Lacaita, A L; Angotzi, G N; Baranauskas, G; Fadiga, L

    2010-01-01

    This paper reports a multi-channel neural recording system-on-chip (SoC) with digital data compression and wireless telemetry. The circuit consists of a 16 amplifiers, an analog time division multiplexer, an 8-bit SAR AD converter, a digital signal processor (DSP) and a wireless narrowband 400-MHz binary FSK transmitter. Even though only 16 amplifiers are present in our current die version, the whole system is designed to work with 64 channels demonstrating the feasibility of a digital processing and narrowband wireless transmission of 64 neural recording channels. A digital data compression, based on the detection of action potentials and storage of correspondent waveforms, allows the use of a 1.25-Mbit/s binary FSK wireless transmission. This moderate bit-rate and a low frequency deviation, Manchester-coded modulation are crucial for exploiting a narrowband wireless link and an efficient embeddable antenna. The chip is realized in a 0.35- εm CMOS process with a power consumption of 105 εW per channel (269 εW per channel with an extended transmission range of 4 m) and an area of 3.1 × 2.7 mm(2). The transmitted signal is captured by a digital TV tuner and demodulated by a wideband phase-locked loop (PLL), and then sent to a PC via an FPGA module. The system has been tested for electrical specifications and its functionality verified in in-vivo neural recording experiments.

  20. Discovery of a novel allosteric inhibitor-binding site in ERK5: comparison with the canonical kinase hinge ATP-binding site

    PubMed Central

    Chen, Hongming; Tucker, Julie; Wang, Xiaotao; Gavine, Paul R.; Phillips, Chris; Augustin, Martin A.; Schreiner, Patrick; Steinbacher, Stefan; Preston, Marian; Ogg, Derek

    2016-01-01

    MAP kinases act as an integration point for multiple biochemical signals and are involved in a wide variety of cellular processes such as proliferation, differentiation, regulation of transcription and development. As a member of the MAP kinase family, ERK5 (MAPK7) is involved in the downstream signalling pathways of various cell-surface receptors, including receptor tyrosine kinases and G protein-coupled receptors. In the current study, five structures of the ERK5 kinase domain co-crystallized with ERK5 inhibitors are reported. Interestingly, three of the compounds bind at a novel allosteric binding site in ERK5, while the other two bind at the typical ATP-binding site. Binding of inhibitors at the allosteric site is accompanied by displacement of the P-loop into the ATP-binding site and is shown to be ATP-competitive in an enzymatic assay of ERK5 kinase activity. Kinase selectivity data show that the most potent allosteric inhibitor exhibits superior kinase selectivity compared with the two inhibitors that bind at the canonical ATP-binding site. An analysis of these structures and comparison with both a previously published ERK5–inhibitor complex structure (PDB entry 4b99) and the structures of three other kinases (CDK2, ITK and MEK) in complex with allosteric inhibitors are presented. PMID:27139631

  1. Allosteric Nanobodies Reveal the Dynamic Range and Diverse Mechanisms of GPCR Activation

    PubMed Central

    Staus, Dean P; Strachan, Ryan T; Manglik, Aashish; Pani, Biswaranjan; Kahsai, Alem W; Kim, Tae Hun; Wingler, Laura M; Ahn, Seungkirl; Chatterjee, Arnab; Masoudi, Ali; Kruse, Andrew C; Pardon, Els; Steyaert, Jan; Weis, William I; Prosser, R. Scott; Kobilka, Brian K; Costa, Tommaso; Lefkowitz, Robert J

    2016-01-01

    G-protein coupled receptors (GPCRs) modulate many physiological processes by transducing a variety of extracellular cues into intracellular responses. Ligand binding to an extracellular orthosteric pocket propagates conformational change to the receptor cytosolic region to promote binding and activation of downstream signaling effectors such as G proteins and β-arrestins. It is widely appreciated that different agonists can share the same binding pocket but evoke unique receptor conformations leading to a wide range of downstream responses (i.e., ‘efficacy’)1. Furthermore, mounting biophysical evidence, primarily using the β-adrenergic receptor (β2AR) as a model system, supports the existence of multiple active and inactive conformational states2–5. However, how agonists with varying efficacy modulate these receptor states to initiate cellular responses is not well understood. Here we report stabilization of two distinct β2AR conformations using single domain camelid antibodies (nanobodies): a previously described positive allosteric nanobody (Nb80) and a newly identified negative allosteric nanobody (Nb60)6,7. We show that Nb60 stabilizes a previously unappreciated low affinity receptor state which corresponds to one of two inactive receptor conformations as delineated by X-ray crystallography and NMR spectroscopy. We find that the agonist isoproterenol has a 15,000-fold higher affinity for the β2AR in the presence of Nb80 compared to Nb60, highlighting the full allosteric range of a GPCR. Assessing the binding of 17 ligands of varying efficacy to the β2AR in the absence and presence of Nb60 or Nb80 reveals large ligand-specific effects that can only be explained using an allosteric model which assumes equilibrium amongst at least three receptor states. Agonists generally exert efficacy by stabilizing the active Nb80-stabilized receptor state (R80). In contrast, for a number of partial agonists, both stabilization of R80 and destabilization of the

  2. Evidence for two different mechanisms triggering the change in quaternary structure of the allosteric enzyme, glucosamine-6-phosphate deaminase.

    PubMed

    Bustos-Jaimes, Ismael; Ramírez-Costa, Montserrat; De Anda-Aguilar, Lorena; Hinojosa-Ocaña, Pilar; Calcagno, Mario L

    2005-02-01

    The generation and propagation of conformational changes associated with ligand binding in the allosteric enzyme glucosamine-6-phosphate deaminase (GlcN6P deaminase, EC 3.5.99.6) from Escherichia coli were analyzed by fluorescence measurements. Single-tryptophan mutant forms of the enzyme were constructed on the basis of previous structural and functional evidence and used as structural-change probes. The reporter residues were placed in the active-site lid (position 174) and in the allosteric site (254 and 234); in addition, signals from the natural Trp residues (15 and 224) were also studied as structural probes. The structural changes produced by the occupation of either the allosteric or the active site by site-specific ligands were monitored through changes in the spectral center of mass (SCM) of their steady-state emission fluorescence spectra. Binding of the allosteric activator produces only minimal signals in titration experiments. In contrast, measurable spectral signals were found when the active site was occupied by a dead-end inhibitor. The results reveal that the two binary complexes, enzyme-activator (R(A)) and enzyme-inhibitor (R(S)) complexes, have structural differences and that they also differ from the ternary complex (R(AS)). The mobility of the active-site lid motif is shown to be independent of the allosteric transition. The active-site ligand induces cooperative SCM changes even in the enzyme-activator complex, indicating that the propagation pathway of the conformational relaxation triggered from the active site is different from that involved in the heterotropic activation. Analysis of the complete set of mutants shows that the occupation of the active site generates structural perturbations, which are propagated to the whole of the monomer and extend to the other subunits. The accumulative effect of these propagated changes should be responsible for the change in the sign of the DeltaG degrees ' of the T to R transition associated with

  3. Suppression of XPM and XPM-induced nonlinear phase noise for RZ-DPSK signals in 40 Gbit/s WDM transmission systems with optimum dispersion mapping.

    PubMed

    Li, Xiaolin; Zhang, Fan; Chen, Zhangyuan; Xu, Anshi

    2007-12-24

    We numerically investigate XPM effect and XPM-induced nonlinear phase noise in both RZ-DPSK and multi-format (RZ-DPSK and RZ-OOK) WDM systems operating at 40 Gbit/s with different dispersion maps. The relative strength of XPM effect and XPM-induced nonlinear phase noise is discussed for both RZ-DPSK and multi-format WDM transmission. With optimum dispersion mapping, XPM and XPM-induced nonlinear phase noise from neighboring channels carrying either OOK or DPSK signals can both be effectively suppressed.

  4. A Novel Allosteric Activator of Free Fatty Acid 2 Receptor Displays Unique Gi-functional Bias*

    PubMed Central

    Bolognini, Daniele; Moss, Catherine E.; Nilsson, Karolina; Petersson, Annika U.; Donnelly, Iona; Sergeev, Eugenia; König, Gabriele M.; Kostenis, Evi; Kurowska-Stolarska, Mariola; Miller, Ashley; Dekker, Niek; Tobin, Andrew B.

    2016-01-01

    The short chain fatty acid receptor FFA2 is able to stimulate signaling via both Gi- and Gq/G11-promoted pathways. These pathways are believed to control distinct physiological end points but FFA2 receptor ligands appropriate to test this hypothesis have been lacking. Herein, we characterize AZ1729, a novel FFA2 regulator that acts as a direct allosteric agonist and as a positive allosteric modulator, increasing the activity of the endogenously produced short chain fatty acid propionate in Gi-mediated pathways, but not at those transduced by Gq/G11. Using AZ1729 in combination with direct inhibitors of Gi and Gq/G11 family G proteins demonstrated that although both arms contribute to propionate-mediated regulation of phospho-ERK1/2 MAP kinase signaling in FFA2-expressing 293 cells, the Gq/G11-mediated pathway is predominant. We extend these studies by employing AZ1729 to dissect physiological FFA2 signaling pathways. The capacity of AZ1729 to act at FFA2 receptors to inhibit β-adrenoreceptor agonist-promoted lipolysis in primary mouse adipocytes and to promote chemotaxis of isolated human neutrophils confirmed these as FFA2 processes mediated by Gi signaling, whereas, in concert with blockade by the Gq/G11 inhibitor FR900359, the inability of AZ1729 to mimic or regulate propionate-mediated release of GLP-1 from mouse colonic preparations defined this physiological response as an end point transduced via activation of Gq/G11. PMID:27385588

  5. Molecular mechanism of the allosteric enhancement of the umami taste sensation.

    PubMed

    Mouritsen, Ole G; Khandelia, Himanshu

    2012-09-01

    The fifth taste quality, umami, arises from binding of glutamate to the umami receptor T1R1/T1R3. The umami taste is enhanced several-fold upon addition of free nucleotides such as guanosine-5'-monophosphate (GMP) to glutamate-containing food. GMP may operate via binding to the ligand-binding domain of the T1R1 part of the umami receptor at an allosteric site. Using molecular dynamics simulations, we show that GMP can stabilize the closed (active) state of T1R1 by binding to the outer vestibule of the so-called Venus flytrap domain of the receptor. The transition between the closed and open conformations was accessed in the simulations. Using principal component analysis, we show that the dynamics of the Venus flytrap domain along the hinge-bending motion that activates signaling is dampened significantly upon binding of glutamate, and further slows down upon binding of GMP at an allosteric site, thus suggesting a molecular mechanism of cooperativity between GMP and glutamate.

  6. Unexpected Allosteric Network Contributes to LRH-1 Co-regulator Selectivity*

    PubMed Central

    Musille, Paul M.; Kossmann, Bradley R.; Kohn, Jeffrey A.; Ivanov, Ivaylo; Ortlund, Eric A.

    2016-01-01

    Phospholipids (PLs) are unusual signaling hormones sensed by the nuclear receptor liver receptor homolog-1 (LRH-1), which has evolved a novel allosteric pathway to support appropriate interaction with co-regulators depending on ligand status. LRH-1 plays an important role in controlling lipid and cholesterol homeostasis and is a potential target for the treatment of metabolic and neoplastic diseases. Although the prospect of modulating LRH-1 via small molecules is exciting, the molecular mechanism linking PL structure to transcriptional co-regulator preference is unknown. Previous studies showed that binding to an activating PL ligand, such as dilauroylphosphatidylcholine, favors LRH-1's interaction with transcriptional co-activators to up-regulate gene expression. Both crystallographic and solution-based structural studies showed that dilauroylphosphatidylcholine binding drives unanticipated structural fluctuations outside of the canonical activation surface in an alternate activation function (AF) region, encompassing the β-sheet-H6 region of the protein. However, the mechanism by which dynamics in the alternate AF influences co-regulator selectivity remains elusive. Here, we pair x-ray crystallography with molecular modeling to identify an unexpected allosteric network that traverses the protein ligand binding pocket and links these two elements to dictate selectivity. We show that communication between the alternate AF region and classical AF2 is correlated with the strength of the co-regulator interaction. This work offers the first glimpse into the conformational dynamics that drive this unusual PL-mediated nuclear hormone receptor activation. PMID:26553876

  7. Allosteric Activation of a G Protein-coupled Receptor with Cell-penetrating Receptor Mimetics*

    PubMed Central

    Zhang, Ping; Leger, Andrew J.; Baleja, James D.; Rana, Rajashree; Corlin, Tiffany; Nguyen, Nga; Koukos, Georgios; Bohm, Andrew; Covic, Lidija; Kuliopulos, Athan

    2015-01-01

    G protein-coupled receptors (GPCRs) are remarkably versatile signaling systems that are activated by a large number of different agonists on the outside of the cell. However, the inside surface of the receptors that couple to G proteins has not yet been effectively modulated for activity or treatment of diseases. Pepducins are cell-penetrating lipopeptides that have enabled chemical and physical access to the intracellular face of GPCRs. The structure of a third intracellular (i3) loop agonist, pepducin, based on protease-activated receptor-1 (PAR1) was solved by NMR and found to closely resemble the i3 loop structure predicted for the intact receptor in the on-state. Mechanistic studies revealed that the pepducin directly interacts with the intracellular H8 helix region of PAR1 and allosterically activates the receptor through the adjacent (D/N)PXXYYY motif through a dimer-like mechanism. The i3 pepducin enhances PAR1/Gα subunit interactions and induces a conformational change in fluorescently labeled PAR1 in a very similar manner to that induced by thrombin. As pepducins can potentially be made to target any GPCR, these data provide insight into the identification of allosteric modulators to this major drug target class. PMID:25934391

  8. Allosteric activation of M4 muscarinic receptors improve behavioral and physiological alterations in early symptomatic YAC128 mice

    PubMed Central

    Pancani, Tristano; Foster, Daniel J.; Moehle, Mark S.; Bichell, Terry Jo; Bradley, Emma; Bridges, Thomas M.; Klar, Rebecca; Poslusney, Mike; Rook, Jerri M.; Daniels, J. Scott; Niswender, Colleen M.; Jones, Carrie K.; Wood, Michael R.; Bowman, Aaron B.; Lindsley, Craig W.; Xiang, Zixiu; Conn, P. Jeffrey

    2015-01-01

    Mutations that lead to Huntington’s disease (HD) result in increased transmission at glutamatergic corticostriatal synapses at early presymptomatic stages that have been postulated to set the stage for pathological changes and symptoms that are observed at later ages. Based on this, pharmacological interventions that reverse excessive corticostriatal transmission may provide a novel approach for reducing early physiological changes and motor symptoms observed in HD. We report that activation of the M4 subtype of muscarinic acetylcholine receptor reduces transmission at corticostriatal synapses and that this effect is dramatically enhanced in presymptomatic YAC128 HD and BACHD relative to wild-type mice. Furthermore, chronic administration of a novel highly selective M4 positive allosteric modulator (PAM) beginning at presymptomatic ages improves motor and synaptic deficits in 5-mo-old YAC128 mice. These data raise the exciting possibility that selective M4 PAMs could provide a therapeutic strategy for the treatment of HD. PMID:26508634

  9. NMR reveals a dynamic allosteric pathway in thrombin

    PubMed Central

    Handley, Lindsey D.; Fuglestad, Brian; Stearns, Kyle; Tonelli, Marco; Fenwick, R. Bryn; Markwick, Phineus R. L.; Komives, Elizabeth A.

    2017-01-01

    Although serine proteases are found ubiquitously in both eukaryotes and prokaryotes, and they comprise the largest of all of the peptidase families, their dynamic motions remain obscure. The backbone dynamics of the coagulation serine protease, apo-thrombin (S195M-thrombin), were compared to the substrate-bound form (PPACK-thrombin). R1, R2, 15N-{1H}NOEs, and relaxation dispersion NMR experiments were measured to capture motions across the ps to ms timescale. The ps-ns motions were not significantly altered upon substrate binding. The relaxation dispersion data revealed that apo-thrombin is highly dynamic, with μs-ms motions throughout the molecule. The region around the N-terminus of the heavy chain, the Na+-binding loop, and the 170 s loop, all of which are implicated in allosteric coupling between effector binding sites and the active site, were dynamic primarily in the apo-form. Most of the loops surrounding the active site become more ordered upon PPACK-binding, but residues in the N-terminal part of the heavy chain, the γ-loop, and anion-binding exosite 1, the main allosteric binding site, retain μs-ms motions. These residues form a dynamic allosteric pathway connecting the active site to the main allosteric site that remains in the substrate-bound form. PMID:28059082

  10. Rational Design of Potent, Small, Synthetic Allosteric Inhibitors of Thrombin

    PubMed Central

    Sidhu, Preetpal Singh; Liang, Aiye; Mehta, Akul Y.; Abdel Aziz, May H.; Zhou, Qibing; Desai, Umesh R.

    2011-01-01

    Thrombin is a key enzyme targeted by the majority of current anticoagulants that are direct inhibitors. Allosteric inhibition of thrombin may offer a major advantage of finely tuned regulation. We present here sulfated benzofurans as the first examples of potent, small allosteric inhibitors of thrombin. A sulfated benzofuran library of 15 sulfated monomers and 13 sulfated dimers with different charged, polar and hydrophobic substituents was studied in this work. Synthesis of the sulfated benzofurans was achieved through a multiple step, highly branched strategy, which culminated with microwave-assisted chemical sulfation. Of the 28 potential inhibitors, eleven exhibited reasonable inhibition of human α-thrombin at pH 7.4. Structure activity relationship analysis indicated that sulfation at the 5-position of the benzofuran scaffold was essential for targeting thrombin. A t-butyl 5-sulfated benzofuran derivative was found to be the most potent thrombin inhibitor with an IC50 of 7.3 μM under physiologically relevant conditions. Michaelis-Menten studies showed an allosteric inhibition phenomenon. Plasma clotting assays indicate that the sulfated benzofurans prolong both the activated partial thromboplastin time and prothrombin time. Overall, this work puts forward sulfated benzofurans as the first small, synthetic molecules as powerful lead compounds for the design of a new class of allosteric inhibitors of thrombin. PMID:21714536

  11. Identification and Quantification of a New Family of Peptide Endocannabinoids (Pepcans) Showing Negative Allosteric Modulation at CB1 Receptors*

    PubMed Central

    Bauer, Mark; Chicca, Andrea; Tamborrini, Marco; Eisen, David; Lerner, Raissa; Lutz, Beat; Poetz, Oliver; Pluschke, Gerd; Gertsch, Jürg

    2012-01-01

    The α-hemoglobin-derived dodecapeptide RVD-hemopressin (RVDPVNFKLLSH) has been proposed to be an endogenous agonist for the cannabinoid receptor type 1 (CB1). To study this peptide, we have raised mAbs against its C-terminal part. Using an immunoaffinity mass spectrometry approach, a whole family of N-terminally extended peptides in addition to RVD-Hpα were identified in rodent brain extracts and human and mouse plasma. We designated these peptides Pepcan-12 (RVDPVNFKLLSH) to Pepcan-23 (SALSDLHAHKLRVDPVNFKLLSH), referring to peptide length. The most abundant Pepcans found in the brain were tested for CB1 receptor binding. In the classical radioligand displacement assay, Pepcan-12 was the most efficacious ligand but only partially displaced both [3H]CP55,940 and [3H]WIN55,212-2. The data were fitted with the allosteric ternary complex model, revealing a cooperativity factor value α < 1, thus indicating a negative allosteric modulation. Dissociation kinetic studies of [3H]CP55,940 in the absence and presence of Pepcan-12 confirmed these results by showing increased dissociation rate constants induced by Pepcan-12. A fluorescently labeled Pepcan-12 analog was synthesized to investigate the binding to CB1 receptors. Competition binding studies revealed Ki values of several Pepcans in the nanomolar range. Accordingly, using competitive ELISA, we found low nanomolar concentrations of Pepcans in human plasma and ∼100 pmol/g in mouse brain. Surprisingly, Pepcan-12 exhibited potent negative allosteric modulation of the orthosteric agonist-induced cAMP accumulation, [35S]GTPγS binding, and CB1 receptor internalization. Pepcans are the first endogenous allosteric modulators identified for CB1 receptors. Given their abundance in the brain, Pepcans could play an important physiological role in modulating endocannabinoid signaling. PMID:22952224

  12. A novel optical single-sideband frequency translation technique for transmission of wireless MIMO signals over fiber-wireless system

    NASA Astrophysics Data System (ADS)

    Shaddad, Redhwan Q.; Mohammad, Abu Bakar; Al-Hetar, Abdulaziz M.; Al-Gailani, Samir A.

    2013-04-01

    The fiber-wireless (FiWi) access network is a powerful hybrid architecture of optical backhaul and wireless front-end to support high data rates and throughput with minimal time delay. By using radio over fiber (ROF) technique, the optical fiber is well adapted to propagate multiple wireless services having different carrier frequencies. However, multiple wireless signals which have the same carrier frequency cannot propagate over a single optical fiber on the same wavelength, such as multi-input multi-output (MIMO) signals. A novel optical single-sideband frequency translation technique is designed and simulated to solve this problem. 240 Mb/s 802.11n MIMO signals are proposed to transport over FiWi system using the proposed approach at 2.4 GHz and 5.0 GHz carrier frequencies. The crosstalk between MIMO signals with the same carrier frequency is excluded, since each MIMO signal is carried on a specific optical wavelength. Error vector magnitude (EVM) values of -29.83 dB (for 2.4 GHz) and -28.41 dB (for 5.0 GHz) have been achieved for bit error rate (BER) 10-5 in the proposed FiWi system.

  13. Porphyrin-encapsulated metal-organic frameworks as mimetic catalysts for electrochemical DNA sensing via allosteric switch of hairpin DNA.

    PubMed

    Ling, Pinghua; Lei, Jianping; Zhang, Lei; Ju, Huangxian

    2015-04-07

    A sensitive electrochemical sensor is designed for DNA detection based on mimetic catalysis of metal-organic framework (MOF) and allosteric switch of hairpin DNA. The functional MOFs are synthesized as signal probes by a one-pot encapsulation of iron(III) meso-5,10,15,20-tetrakis(4-carboxyphenyl) porphyrin chloride (FeTCPP) into a prototypal MOF, HKUST-1(Cu), and sequentially conjugated with streptavidin (SA) as a recognition element. The resulting FeTCPP@MOF composites can mimetically catalyze the oxidation of o-phenylenediamine (o-PD) to 2,2'-diaminoazobenzene, which is a good electrochemical indicator for signal readout. The presence of target DNA introduces the allosteric switch of hairpin DNA to form SA aptamer, and thus, FeTCPP@MOF-SA probe is brought on the electrode surface via the specific recognition between SA and the corresponding aptamer, resulting in the enhancement of electrochemical signal. The "signal-on" electrochemical sensor can detect target DNA down to 0.48 fM with the linear range of 10 fM to 10 nM. Moreover, the MOF-based electrochemical sensor exhibits acceptable selectivity against even a single mismatched DNA and good feasibility in complex serum matrixes. This strategy opens up a new direction of porphyrin-functionalized MOF for signal transduction in electrochemical biosensing.

  14. Allosteric mechanisms can be distinguished using structural mass spectrometry

    PubMed Central

    Dyachenko, Andrey; Gruber, Ranit; Shimon, Liat; Horovitz, Amnon; Sharon, Michal

    2013-01-01

    The activity of many proteins, including metabolic enzymes, molecular machines, and ion channels, is often regulated by conformational changes that are induced or stabilized by ligand binding. In cases of multimeric proteins, such allosteric regulation has often been described by the concerted Monod–Wyman–Changeux and sequential Koshland–Némethy–Filmer classic models of cooperativity. Despite the important functional implications of the mechanism of cooperativity, it has been impossible in many cases to distinguish between these various allosteric models using ensemble measurements of ligand binding in bulk protein solutions. Here, we demonstrate that structural MS offers a way to break this impasse by providing the full distribution of ligand-bound states of a protein complex. Given this distribution, it is possible to determine all the binding constants of a ligand to a highly multimeric cooperative system, and thereby infer its allosteric mechanism. Our approach to the dissection of allosteric mechanisms relies on advances in MS—which provide the required resolution of ligand-bound states—and in data analysis. We validated our approach using the well-characterized Escherichia coli chaperone GroEL, a double-heptameric ring containing 14 ATP binding sites, which has become a paradigm for molecular machines. The values of the 14 binding constants of ATP to GroEL were determined, and the ATP-loading pathway of the chaperone was characterized. The methodology and analyses presented here are directly applicable to numerous other cooperative systems and are therefore expected to promote further research on allosteric systems. PMID:23589876

  15. Maturation of glutamatergic transmission in the vestibulo-olivary pathway impacts on the registration of head rotational signals in the brainstem of rats.

    PubMed

    Lai, Chun-Hong; Ma, Chun-Wai; Lai, Suk-King; Han, Lei; Wong, Hoi-Man; Yeung, Kelvin Wai-Kwok; Shum, Daisy Kwok-Yan; Chan, Ying-Shing

    2016-01-01

    The recognition of head orientation in the adult involves multi-level integration of inputs within the central vestibular circuitry. How the different inputs are recruited during postnatal development remains unclear. We hypothesize that glutamatergic transmission at the vestibular nucleus contributes to developmental registration of head orientations along the vestibulo-olivary pathway. To investigate the maturation profile by which head rotational signals are registered in the brainstem, we used sinusoidal rotations on the orthogonal planes of the three pairs of semicircular canals. Fos expression was used as readout of neurons responsive to the rotational stimulus. Neurons in the vestibular nucleus and prepositus hypoglossal nucleus responded to all rotations as early as P4 and reached adult numbers by P21. In the reticular formation and inferior olive, neurons also responded to horizontal rotations as early as P4 but to vertical rotations not until P21 and P25, respectively. Neuronal subpopulations that distinguish between rotations activating the orthogonally oriented vertical canals were identifiable in the medial and spinal vestibular nuclei by P14 and in the inferior olivary subnuclei IOβ and IOK by P25. Neonatal perturbation of glutamate transmission in the vestibular nucleus was sufficient to derange formation of this distribution in the inferior olive. This is the first demonstration that developmental refinement of glutamatergic synapses in the central vestibular circuitry is essential for developmental registration of head rotational signals in the brainstem.

  16. Transmission of 107-Gb/s mode and polarization multiplexed CO-OFDM signal over a two-mode fiber.

    PubMed

    Li, An; Al Amin, Abdullah; Chen, Xi; Shieh, William

    2011-04-25

    In addition to the dimensions of time, frequency, complex constellation, and polarization, spatial mode can be the fifth dimension to be explored for modulation and multiplexing in optical fiber communications. In this paper, we demonstrate successful transmission of 107-Gb/s dual-mode and dual-polarization coherent optical orthogonal frequency-division multiplexing (CO-OFDM) over a 4.5-km two-mode fiber. A mechanically-induced LP01/LP11 mode converter is used as the mode selective element in a spatial-mode multiplexed system.

  17. A unique method to study acoustic transmission through ducts using signal synthesis and averaging of acoustic pulses

    NASA Technical Reports Server (NTRS)

    Salikuddin, M.; Ramakrishnan, R.; Ahuja, K. K.; Brown, W. H.

    1981-01-01

    An acoustic impulse technique using a loudspeaker driver is developed to measure the acoustic properties of a duct/nozzle system. A signal synthesis method is used to generate a desired single pulse with a flat spectrum. The convolution of the desired signal and the inverse Fourier transform of the reciprocal of the driver's response are then fed to the driver. A signal averaging process eliminates the jet mixing noise from the mixture of jet noise and the internal noise, thereby allowing very low intensity signals to be measured accurately, even for high velocity jets. A theoretical analysis is carried out to predict the incident sound field; this is used to help determine the number and locations of the induct measurement points to account for the contributions due to higher order modes present in the incident tube method. The impulse technique is validated by comparing experimentally determined acoustic characteristics of a duct-nozzle system with similar results obtained by the impedance tube method. Absolute agreement in the comparisons was poor, but the overall shapes of the time histories and spectral distributions were much alike.

  18. Improvement of Information Transmission of Suprathreshold Input Signal with Stochastic Resonance in Hippocampal CA1 Neuron Network

    NASA Astrophysics Data System (ADS)

    Kawaguchi, Minato; Mino, Hiroyuki; Momose, Keiko; Durand, Dominique M.

    We investigate if and how SR (stochastic resonance) can be shown in the presence of supra-threshold signals (SSR) in physiologically realistic neural networks. The mutual information was maximized at a specific amplitude of noise in larger neural networks, implying SSR.

  19. Involvement of BDNF signaling transmission from basolateral amygdala to infralimbic prefrontal cortex in conditioned taste aversion extinction.

    PubMed

    Xin, Jian; Ma, Ling; Zhang, Tian-Yi; Yu, Hui; Wang, Yue; Kong, Liang; Chen, Zhe-Yu

    2014-05-21

    Brain-derived neurotrophic factor (BDNF) and its receptor, tropomyosin-related kinase receptor B (TrkB), play a critical role in memory extinction. However, the detailed role of BDNF in memory extinction on the basis of neural circuit has not been fully understood. Here, we aim to investigate the role of BDNF signaling circuit in mediating conditioned taste aversion (CTA) memory extinction of the rats. We found region-specific changes in BDNF gene expression during CTA extinction. CTA extinction led to increased BDNF gene expression in the basolateral amygdala (BLA) and infralimbic prefrontal cortex (IL) but not in the central amygdaloid nucleus (CeA) and hippocampus (HIP). Moreover, blocking BDNF signaling or exogenous microinjection of BDNF into the BLA or IL could disrupt or enhance CTA extinction, which suggested that BDNF signaling in the BLA and IL is necessary and sufficient for CTA extinction. Interestingly, we found that microinjection of BDNF-neutralizing antibody into the BLA could abolish the extinction training-induced BDNF mRNA level increase in the IL, but not vice versa, demonstrating that BDNF signaling is transmitted from the BLA to IL during extinction. Finally, the accelerated extinction learning by infusion of exogenous BDNF in the BLA could also be blocked by IL infusion of BDNF-neutralizing antibody rather than vice versa, indicating that the IL, but not BLA, is the primary action site of BDNF in CTA extinction. Together, these data suggest that BLA-IL circuit regulates CTA memory extinction by identifying BDNF as a key regulator.

  20. Local neuropeptide signaling modulates serotonergic transmission to shape the temporal organization of C. elegans egg-laying behavior.

    PubMed

    Banerjee, Navonil; Bhattacharya, Raja; Gorczyca, Michael; Collins, Kevin M; Francis, Michael M

    2017-04-06

    Animal behaviors are often composed of distinct alternating behavioral states. Neuromodulatory signals are thought to be critical for establishing stable behavioral states and for orchestrating transitions between them. However, we have only a limited understanding of how neuromodulatory systems act in vivo to alter circuit performance and shape behavior. To address these questions, we have investigated neuromodulatory signaling in the context of Caenorhabditis elegans egg-laying. Egg-laying activity cycles between discrete states-short bursts of egg deposition (active phases) that alternate with prolonged quiescent periods (inactive phases). Here using genetic, pharmacological and optogenetic approaches for cell-specific activation and inhibition, we show that a group of neurosecretory cells (uv1) located in close spatial proximity to the egg-laying neuromusculature direct the temporal organization of egg-laying by prolonging the duration of inactive phases. We demonstrate that the modulatory effects of the uv1 cells are mediated by peptides encoded by the nlp-7 and flp-11 genes that act locally to inhibit circuit activity, primarily by inhibiting vesicular release of serotonin from HSN motor neurons. This peptidergic inhibition is achieved, at least in part, by reducing synaptic vesicle abundance in the HSN motor neurons. By linking the in vivo actions of specific neuropeptide signaling systems with the generation of stable behavioral outcomes, our study reveals how cycles of neuromodulation emanating from non-neuronal cells can fundamentally shape the organization of a behavioral program.

  1. Glucocorticoids Regulate Glutamate and GABA Synapse-Specific Retrograde Transmission via Divergent Non-Genomic Signaling Pathways

    PubMed Central

    Di, Shi; Maxson, Marc M.; Franco, Alier; Tasker, Jeffrey G.

    2009-01-01

    Glucocorticoids exert an opposing rapid regulation of glutamate and GABA synaptic inputs to hypothalamic magnocellular neurons via the activation of postsynaptic membrane-associated receptors and the release of retrograde messengers. Glucocorticoids suppress synaptic glutamate release via the retrograde release of endocannabinoids and facilitate synaptic GABA release via an unknown retrograde messenger. Here, we show that the glucocorticoid facilitation of GABA inputs is due to the retrograde release of neuronal nitric oxide, and that glucocorticoid-induced endocannabinoid synthesis and nitric oxide synthesis are mediated by divergent G protein signaling mechanisms. While the glucocorticoid-induced, endocannabinoid-mediated suppression of glutamate release is dependent on activation of the Gαs G protein subunit and cAMP-PKA activation, the nitric oxide facilitation of GABA release is mediated by Gβγ signaling that leads to activation of neuronal nitric oxide synthase. Our findings indicate, therefore, that glucocorticoids exert opposing rapid actions on glutamate and GABA release by activating divergent G protein signaling pathways that trigger the synthesis of, and glutamate and GABA synapse-specific retrograde actions of, endocannabinoids and nitric oxide, respectively. The simultaneous rapid stimulation of nitric oxide and endocannabinoid synthesis by glucocorticoids has important implications for the impact of stress on the brain as well as on neural-immune interactions in the hypothalamus. PMID:19144839

  2. Roles of gap junctions and hemichannels in bone cell functions and in signal transmission of mechanical stress

    PubMed Central

    Jiang, Jean Xin; Siller-Jackson, Arlene Janel; Burra, Sirisha

    2007-01-01

    Gap junctions formed by connexins (Cx) play an important role in transmitting signals between bone cells such as osteoblasts and osteoclasts, cells responsible for bone formation and bone remodeling, respectively. Gap junction intercellular communication (GJIC) has been demonstrated to mediate the process of osteoblast differentiation and bone formation. Furthermore, GJIC propagates Ca2+ signaling, conveys anabolic effects of hormones and growth factors, and regulates gene transcription of osteoblast differentiation markers. GJIC is also implicated to regulate osteoclast formation, survival and apoptosis. Compared with other bone cells, the most abundant type are osteocytes, which express large amounts of connexins. Mechanosensing osteocytes connect and form gap junctions with themselves and other cells only through the tips of their dendritic processes, a relatively small percent of the total cell surface area compared to other cells. Recent studies show that in addition to gap junctions, osteoblasts and osteocytes express functional hemichannels, the un-opposed halves of gap junction channels. Hemichannels are localized at the cell surface and function independently of gap junctions. Hemichannels in osteocytes mediate the immediate release of prostaglandins in response to mechanical stress. The major challenges remaining in the field are how the functions of these two types of channels are coordinated in bone cells and what the asserted, distinct effects of these channels are on bone formation and remodeling processes, and on conveying signals elicited by mechanical loading. PMID:17127393

  3. Roles of gap junctions and hemichannels in bone cell functions and in signal transmission of mechanical stress.

    PubMed

    Jiang, Jean Xin; Siller-Jackson, Arlene Janel; Burra, Sirisha

    2007-01-01

    Gap junctions formed by connexins (Cx) play an important role in transmitting signals between bone cells such as osteoblasts and osteoclasts, cells responsible for bone formation and bone remodeling, respectively. Gap junction intercellular communication (GJIC) has been demonstrated to mediate the process of osteoblast differentiation and bone formation. Furthermore, GJIC propagates Ca2+ signaling, conveys anabolic effects of hormones and growth factors, and regulates gene transcription of osteoblast differentiation markers. GJIC is also implicated to regulate osteoclast formation, survival and apoptosis. Compared with other bone cells, the most abundant type are osteocytes, which express large amounts of connexins. Mechanosensing osteocytes connect and form gap junctions with themselves and other cells only through the tips of their dendritic processes, a relatively small percent of the total cell surface area compared to other cells. Recent studies show that in addition to gap junctions, osteoblasts and osteocytes express functional hemichannels, the un-opposed halves of gap junction channels. Hemichannels are localized at the cell surface and function independently of gap junctions. Hemichannels in osteocytes mediate the immediate release of prostaglandins in response to mechanical stress. The major challenges remaining in the field are how the functions of these two types of channels are coordinated in bone cells and what the asserted, distinct effects of these channels are on bone formation and remodeling processes, and on conveying signals elicited by mechanical loading.

  4. Characterization of the PAS domain in the sensor-kinase BvgS: mechanical role in signal transmission

    PubMed Central

    2013-01-01

    Background In bacteria, signal-transduction two-component systems are major players for adaptation to environmental stimuli. The perception of a chemical or physical signal by a sensor-kinase triggers its autophosphorylation. The phosphoryl group is then transferred to the cognate response regulator, which mediates the appropriate adaptive response. Virulence of the whooping cough agent Bordetella pertussis is controlled by the two-component system BvgAS. Atypically, the sensor-kinase BvgS is active without specific stimuli at 37°C in laboratory conditions and is inactivated by the addition of negative chemical modulators. The structure of BvgS is complex, with two tandem periplasmic Venus flytrap domains and a cytoplasmic PAS domain that precedes the kinase domain, which is followed by additional phosphotransfer domains. PAS domains are small, ubiquitous sensing or regulatory domains. The function of the PAS domain in BvgS remains unknown. Results We showed that recombinant BvgS PAS proteins form dimers that are stabilized by α helical regions flanking the PAS core. A structural model of the PAS domain dimer was built and probed by site-directed mutagenesis and by biochemical and functional analyses. Although we found no ligands for the PAS domain cavity, its integrity is required for signaling. We also showed that the structural stability of the PAS core and its proper coupling to its flanking N- and C-terminal α helices are crucial for BvgS activity. Conclusions We propose that a major function of the BvgS PAS domain is to maintain conformational signals arising from mechanical strain generated by the periplasmic domain. The tight structure of the PAS core and its connections with the upstream and downstream helices ensure signaling to the kinase domain, which determines BvgS activity. Many mild substitutions that map to the PAS domain keep BvgS active but make it unresponsive to negative modulators, supporting that modulation increases conformational strain

  5. Investigation of in-band transmission of both spectral amplitude coding/optical code division multiple-access and wavelength division multiplexing signals

    NASA Astrophysics Data System (ADS)

    Ashour, Isaac A. M.; Shaari, Sahbudin; Shalaby, Hossam M. H.; Menon, P. Susthitha

    2011-06-01

    The transmission of both optical code division multiple-access (OCDMA) and wavelength division multiplexing (WDM) users on the same band is investigated. Code pulses of spectral amplitude coding (SAC)/optical code division multiple-access (CDMA) are overlaid onto a multichannel WDM system. Notch filters are utilized in order to suppress the WDM interference signals for detection of optical broadband CDMA signals. Modified quadratic congruence (MQC) codes are used as the signature codes for the SAC/OCDMA system. The proposed system is simulated and its performance in terms of both the bit-error rate and Q-factor are determined. In addition, eavesdropper probability of error-free code detection is evaluated. Our results are compared to traditional nonhybrid systems. It is concluded that the proposed hybrid scheme still achieves acceptable performance. In addition, it provides enhanced data confidentiality as compared to the scheme with SAC/OCDMA only. It is also shown that the performance of the proposed system is limited by the interference of the WDM signals. Furthermore, the simulation illustrates the tradeoff between the performance and confidentiality for authorized users.

  6. Molecular mechanism of allosteric substrate activation in a thiamine diphosphate-dependent decarboxylase.

    PubMed

    Versées, Wim; Spaepen, Stijn; Wood, Martin D H; Leeper, Finian J; Vanderleyden, Jos; Steyaert, Jan

    2007-11-30

    Thiamine diphosphate-dependent enzymes are involved in a wide variety of metabolic pathways. The molecular mechanism behind active site communication and substrate activation, observed in some of these enzymes, has since long been an area of debate. Here, we report the crystal structures of a phenylpyruvate decarboxylase in complex with its substrates and a covalent reaction intermediate analogue. These structures reveal the regulatory site and unveil the mechanism of allosteric substrate activation. This signal transduction relies on quaternary structure reorganizations, domain rotations, and a pathway of local conformational changes that are relayed from the regulatory site to the active site. The current findings thus uncover the molecular mechanism by which the binding of a substrate in the regulatory site is linked to the mounting of the catalytic machinery in the active site in this thiamine diphosphate-dependent enzyme.

  7. Evidence for allosteric interactions of antagonist binding to the smoothened receptor.

    PubMed

    Rominger, Cynthia M; Bee, Wei-Lin Tiger; Copeland, Robert A; Davenport, Elizabeth A; Gilmartin, Aidan; Gontarek, Richard; Hornberger, Keith R; Kallal, Lorena A; Lai, Zhihong; Lawrie, Kenneth; Lu, Quinn; McMillan, Lynette; Truong, Maggie; Tummino, Peter J; Turunen, Brandon; Will, Matthew; Zuercher, William J; Rominger, David H

    2009-06-01

    The Smoothened receptor (Smo) mediates hedgehog (Hh) signaling critical for development, cell growth, and migration, as well as stem cell maintenance. Aberrant Hh signaling pathway activation has been implicated in a variety of cancers, and small-molecule antagonists of Smo have entered human clinical trials for the treatment of cancer. Here, we report the biochemical characterization of allosteric interactions of agonists and antagonists for Smo. Binding of two radioligands, [(3)H]3-chloro-N-[trans-4-(methylamino)cyclohexyl]-N-{[3-(4-pyridinyl)-phenyl]methyl}-1-benzothiophene-2-carboxamide (SAG-1.3) (agonist) and [(3)H]cyclopamine (antagonist), was characterized using human Smo expressed in human embryonic kidney 293F membranes. We observed full displacement of [(3)H]cyclopamine by all Smo agonist and antagonist ligands examined. N-[(1E)-(3,5-Dimethyl-1-phenyl-1H-pyrazol-4-yl)methylidene]-4-(phenylmethyl)-1-piperazinamine (SANT-1), an antagonist, did not fully inhibit the binding of [(3)H]SAG-1.3. In a functional cell-based beta-lactamase reporter gene assay, SANT-1 and N-[3-(1H-benzimidazol-2-yl)-4-chlorophenyl]-3,4,5-tris(ethyloxy)-benzamide (SANT-2) fully inhibited 3-chloro-4,7-difluoro-N-[trans-4-(methylamino)cyclohexyl]-N-{[3-(4-pyridinyl)phenyl]methyl}-1-benzothiophene-2-carboxamide (SAG-1.5)-induced Hh pathway activation. Detailed "Schild-type" radioligand binding analysis with [(3)H]SAG-1.3 revealed that two structurally distinct Smoothened receptor antagonists, SANT-1 and SANT-2, bound in a manner consistent with that of allosteric modulation. Our mechanism of action characterization of radioligand binding to Smo combined with functional data provides a better understanding of small-molecule interactions with Smo and their influence on the Hh pathway.

  8. Identification of an antithrombotic allosteric modulator that acts through helix 8 of PAR1

    PubMed Central

    Dowal, Louisa; Sim, Derek S.; Dilks, James R.; Blair, Price; Beaudry, Sarah; Denker, Bradley M.; Koukos, Georgios; Kuliopulos, Athan; Flaumenhaft, Robert

    2011-01-01

    G protein-coupled receptors (GPCRs) can assume multiple conformations and possess multiple binding sites. Whereas endogenous agonists acting at the orthosteric binding site stabilize the active receptor conformation, small molecules that act at nonorthosteric sites can stabilize alternative conformations. The large majority of these allosteric modulators associate with extracellular loops of GPCRs. The role of intracellular domains in mediating allosteric modulation is largely unknown. In screening a small-molecule library for inhibitors of platelet activation, we identified a family of compounds that modified PAR1-mediated granule secretion. The most potent inhibitory compound, termed JF5, also demonstrated noncompetitive inhibition of the α2A-adrenergic receptor. Aggregation studies using a battery of platelet GPCR agonists demonstrated that sensitivity to JF5 was limited to GPCRs that possessed a constrained eighth helix, as defined by a C-terminal palmitoylation site and interactions with TM7 and the i1 loop. Inhibition by JF5 was overcome in a PAR1 mutant in which the eighth helix was deleted, confirming a role for helix 8 in JF5 activity. Evaluation of downstream signaling showed that JF5 was selective with regard to G protein coupling, blocking signaling mediated by Gαq but not Gα12. The compound inhibited thrombus formation in vivo following vascular injury with an IC50 of ∼1 mg/kg. These results indicate a role for helix 8 in conferring sensitivity to small molecules, and show that this sensitivity can be exploited to control platelet activation during thrombus formation. PMID:21282664

  9. Allosteric Modulation of Hormone Release from Thyroxine and Corticosteroid-binding Globulins*

    PubMed Central

    Qi, Xiaoqiang; Loiseau, François; Chan, Wee Lee; Yan, Yahui; Wei, Zhenquan; Milroy, Lech-Gustav; Myers, Rebecca M.; Ley, Steven V.; Read, Randy J.; Carrell, Robin W.; Zhou, Aiwu

    2011-01-01

    The release of hormones from thyroxine-binding globulin (TBG) and corticosteroid-binding globulin (CBG) is regulated by movement of the reactive center loop in and out of the β-sheet A of the molecule. To investigate how these changes are transmitted to the hormone-binding site, we developed a sensitive assay using a synthesized thyroxine fluorophore and solved the crystal structures of reactive loop cleaved TBG together with its complexes with thyroxine, the thyroxine fluorophores, furosemide, and mefenamic acid. Cleavage of the reactive loop results in its complete insertion into the β-sheet A and a substantial but incomplete decrease in binding affinity in both TBG and CBG. We show here that the direct interaction between residue Thr342 of the reactive loop and Tyr241 of the hormone binding site contributes to thyroxine binding and release following reactive loop insertion. However, a much larger effect occurs allosterically due to stretching of the connecting loop to the top of the D helix (hD), as confirmed in TBG with shortening of the loop by three residues, making it insensitive to the S-to-R transition. The transmission of the changes in the hD loop to the binding pocket is seen to involve coherent movements in the s2/3B loop linked to the hD loop by Lys243, which is, in turn, linked to the s4/5B loop, flanking the thyroxine-binding site, by Arg378. Overall, the coordinated movements of the reactive loop, hD, and the hormone binding site allow the allosteric regulation of hormone release, as with the modulation demonstrated here in response to changes in temperature. PMID:21325280

  10. Allosteric Pathways in the PPARγ-RXRα nuclear receptor complex

    NASA Astrophysics Data System (ADS)

    Ricci, Clarisse G.; Silveira, Rodrigo L.; Rivalta, Ivan; Batista, Victor S.; Skaf, Munir S.

    2016-01-01

    Understanding the nature of allostery in DNA-nuclear receptor (NR) complexes is of fundamental importance for drug development since NRs regulate the transcription of a myriad of genes in humans and other metazoans. Here, we investigate allostery in the peroxisome proliferator-activated/retinoid X receptor heterodimer. This important NR complex is a target for antidiabetic drugs since it binds to DNA and functions as a transcription factor essential for insulin sensitization and lipid metabolism. We find evidence of interdependent motions of Ω-loops and PPARγ-DNA binding domain with contacts susceptible to conformational changes and mutations, critical for regulating transcriptional functions in response to sequence-dependent DNA dynamics. Statistical network analysis of the correlated motions, observed in molecular dynamics simulations, shows preferential allosteric pathways with convergence centers comprised of polar amino acid residues. These findings are particularly relevant for the design of allosteric modulators of ligand-dependent transcription factors.

  11. Allosterism and Structure in Thermally Activated Transient Receptor Potential Channels.

    PubMed

    Diaz-Franulic, Ignacio; Poblete, Horacio; Miño-Galaz, Germán; González, Carlos; Latorre, Ramón

    2016-07-05

    The molecular sensors that mediate temperature changes in living organisms are a large family of proteins known as thermosensitive transient receptor potential (TRP) ion channels. These membrane proteins are polymodal receptors that can be activated by cold or hot temperatures, depending on the channel subtype, voltage, and ligands. The stimuli sensors are allosterically coupled to a pore domain, increasing the probability of finding the channel in its ion conductive conformation. In this review we first discuss the allosteric coupling between the temperature and voltage sensor modules and the pore domain, and then discuss the thermodynamic foundations of thermo-TRP channel activation. We provide a structural overview of the molecular determinants of temperature sensing. We also posit an anisotropic thermal diffusion model that may explain the large temperature sensitivity of TRP channels. Additionally, we examine the effect of several ligands on TRP channel function and the evidence regarding their mechanisms of action.

  12. Allosteric Pathways in the PPARγ-RXRα nuclear receptor complex

    PubMed Central

    Ricci, Clarisse G.; Silveira, Rodrigo L.; Rivalta, Ivan; Batista, Victor S.; Skaf, Munir S.

    2016-01-01

    Understanding the nature of allostery in DNA-nuclear receptor (NR) complexes is of fundamental importance for drug development since NRs regulate the transcription of a myriad of genes in humans and other metazoans. Here, we investigate allostery in the peroxisome proliferator-activated/retinoid X receptor heterodimer. This important NR complex is a target for antidiabetic drugs since it binds to DNA and functions as a transcription factor essential for insulin sensitization and lipid metabolism. We find evidence of interdependent motions of Ω-loops and PPARγ-DNA binding domain with contacts susceptible to conformational changes and mutations, critical for regulating transcriptional functions in response to sequence-dependent DNA dynamics. Statistical network analysis of the correlated motions, observed in molecular dynamics simulations, shows preferential allosteric pathways with convergence centers comprised of polar amino acid residues. These findings are particularly relevant for the design of allosteric modulators of ligand-dependent transcription factors. PMID:26823026

  13. Allosteric mechanisms of nuclear receptors: insights from computational simulations.

    PubMed

    Mackinnon, Jonathan A G; Gallastegui, Nerea; Osguthorpe, David J; Hagler, Arnold T; Estébanez-Perpiñá, Eva

    2014-08-05

    The traditional structural view of allostery defines this key regulatory mechanism as the ability of one conformational event (allosteric site) to initiate another in a separate location (active site). In recent years computational simulations conducted to understand how this phenomenon occurs in nuclear receptors (NRs) has gained significant traction. These results have yield insights into allosteric changes and communication mechanisms that underpin ligand binding, coactivator binding site formation, post-translational modifications, and oncogenic mutations. Moreover, substantial efforts have been made in understanding the dynamic processes involved in ligand binding and coregulator recruitment to different NR conformations in order to predict cell/tissue-selective pharmacological outcomes of drugs. They also have improved the accuracy of in silico screening protocols so that nowadays they are becoming part of optimisation protocols for novel therapeutics. Here we summarise the important contributions that computational simulations have made towards understanding the structure/function relationships of NRs and how these can be exploited for rational drug design.

  14. Ensemble Properties of Network Rigidity Reveal Allosteric Mechanisms

    PubMed Central

    Jacobs, Donald J.; Livesay, Dennis R.; Mottonen, James M.; Vorov, Oleg K.; Istomin, Andrei Y.; Verma, Deeptak

    2015-01-01

    The distance constraint model (DCM) is a unique computational modeling paradigm that integrates mechanical and thermodynamic descriptions of macromolecular structure. That is, network rigidity calculations are used to account for nonadditivity within entropy components, thus restoring the utility of free energy decomposition. The DCM outputs a large number of structural characterizations that collectively allow for quantified stability/flexibility relationships (QSFR) to be identified. In this review, we describe the theoretical underpinnings of the DCM and introduce several common QSFR metrics. Application of the DCM across protein families highlights the sensitivity within the set of protein structure residue-to-residue couplings. Further, we have developed a perturbation method to identify putative allosteric sites, where large changes in QSFR upon rigidification (mimicking ligand-binding) detect sites likely to invoke allosteric changes. PMID:22052496

  15. OPTICAL RECORDING AND COMMUNICATION: Mismatch of the pump parameters of coupled chaotic lasers and the signal recovery errors in the data transmission scheme based on these lasers

    NASA Astrophysics Data System (ADS)

    Ledenev, V. I.

    2004-08-01

    The pulse characteristics of a periodically pumped chaotic CO2 laser are calculated numerically. The errors of the encoding-function recovery in the data transmission link based on two optically coupled chaotic CO2 lasers are studied depending on the amplitude mismatch and pump modulation frequencies. It is shown that the recovery error is minimal when the pump parameters of a transmitter and a receiver are identical. It is also shown that the mismatch of the pump amplitudes of the receiver and transmitter affects the recovery error much weaker than the mismatch of their periods. The dependences of the recovery error on the signal modulation degree are found. It is found that the shape of the recovered encoding function is determined by the type of a deviated parameter.

  16. Single-lane 180  Gb/s DB-PAM-4-signal transmission over an 80  km DCF-free SSMF link.

    PubMed

    Zhang, Qiang; Stojanovic, Nebojsa; Wei, Jinlong; Xie, Changsong

    2017-02-15

    We experimentally demonstrate the generation and successful transmission of a single-lane 180 Gb/s (90 GBd) duo-binary four-level pulse-amplitude modulation (DB-PAM-4) signal over an 80 km standard single-mode fiber with a net data-rate of 150 Gb/s, enabled by a dual-drive Mach-Zehnder modulator-aided dispersion pre-compensation. A net data rate of 168 Gb/s is also achievable, for ranges up to 40 km. To mitigate bandwidth limitations and error spreading, pre-coded DB-PAM-4 is used; the pre-coding provides a gain of approximately 1 dB. A Volterra filter and a maximum likelihood sequence estimator are used at the receiver side to reduce the linear and nonlinear distortions.

  17. Novel bivalent positive allosteric modulators of AMPA receptor.

    PubMed

    Lavrov, M I; Grigor'ev, V V; Bachurin, S O; Palyulin, V A; Zefirov, N S

    2015-01-01

    A positive allosteric modulator of AMPA receptors has been designed using computer-aided molecular modeling techniques. It possessed a record high experimentally confirmed potency in the picomolar concentration range and belongs to a new type of bivalent AMPA receptor ligands containing bicyclo[3.3.1]nonane scaffold. The suggested structure could serve as a basis for further optimization and development of drugs for the treatment of neurodegenerative diseases, cognition enhancement, and improvement of memory.

  18. Micro- and nano-structural details of a spider's filter for substrate vibrations: relevance for low-frequency signal transmission

    PubMed Central

    Erko, Maxim; Younes-Metzler, Osnat; Rack, Alexander; Zaslansky, Paul; Young, Seth L.; Milliron, Garrett; Chyasnavichyus, Marius; Barth, Friedrich G.; Fratzl, Peter; Tsukruk, Vladimir; Zlotnikov, Igor; Politi, Yael

    2015-01-01

    The metatarsal lyriform organ of the Central American wandering spider Cupiennius salei is its most sensitive vibration detector. It is able to sense a wide range of vibration stimuli over four orders of magnitude in frequency between at least as low as 0.1 Hz and several kilohertz. Transmission of the vibrations to the slit organ is controlled by a cuticular pad in front of it. While the mechanism of high-frequency stimulus transfer (above ca 40 Hz) is well understood and related to the viscoelastic properties of the pad's epicuticle, it is not yet clear how low-frequency stimuli (less than 40 Hz) are transmitted. Here, we study how the pad material affects the pad's mechanical properties and thus its role in the transfer of the stimulus, using a variety of experimental techniques, such as X-ray micro-computed tomography for three-dimensional imaging, X-ray scattering for structural analysis, and atomic force microscopy and scanning electron microscopy for surface imaging. The mechanical properties were investigated using scanning acoustic microscopy and nanoindentation. We show that large tarsal deflections cause large deformation in the distal highly hydrated part of the pad. Beyond this region, a sclerotized region serves as a supporting frame which resists the deformation and is displaced to push against the slits, with displacement values considerably scaled down to only a few micrometres. Unravelling the structural arrangement in such specialized structures may provide conceptual ideas for the design of new materials capable of controlling a technical sensor's specificity and selectivity, which is so typical of biological sensors. PMID:25631567

  19. Discovery of a Negative Allosteric Modulator of GABAB Receptors

    PubMed Central

    2014-01-01

    Initialized from the scaffold of CGP7930, an allosteric agonist of GABAB receptors, a series of noncompetitive antagonists were discovered. Among these compounds, compounds 3, 6, and 14 decreased agonist GABA-induced maximal effect of IP3 production in HEK293 cells overexpressing GABAB receptors and Gqi9 proteins without changing the EC50. Compounds 3, 6, and 14 not only inhibited agonist baclofen-induced ERK1/2 phosphorylation but also blocked CGP7930-induced ERK1/2 phosphorylation in HEK293 cells overexpressing GABAB receptors. The results suggested that compounds 3, 6, and 14 are negative allosteric modulators of GABAB receptors. The representative compound 14 decreased GABA-induced IP3 production with IC50 of 37.9 μM and had no effect on other GPCR Class C members such as mGluR1, mGluR2, and mGluR5. Finally, we showed that compound 14 did not bind to the orthosteric binding sites of GABAB receptors, demonstrating that compound 14 negatively modulated GABAB receptors activity as a negative allosteric modulator. PMID:25050158

  20. Zinc as Allosteric Ion Channel Modulator: Ionotropic Receptors as Metalloproteins.

    PubMed

    Peralta, Francisco Andrés; Huidobro-Toro, Juan Pablo

    2016-07-02

    Zinc is an essential metal to life. This transition metal is a structural component of many proteins and is actively involved in the catalytic activity of cell enzymes. In either case, these zinc-containing proteins are metalloproteins. However, the amino acid residues that serve as ligands for metal coordination are not necessarily the same in structural proteins compared to enzymes. While crystals of structural proteins that bind zinc reveal a higher preference for cysteine sulfhydryls rather than histidine imidazole rings, catalytic enzymes reveal the opposite, i.e., a greater preference for the histidines over cysteines for catalysis, plus the influence of carboxylic acids. Based on this paradigm, we reviewed the putative ligands of zinc in ionotropic receptors, where zinc has been described as an allosteric modulator of channel receptors. Although these receptors do not strictly qualify as metalloproteins since they do not normally bind zinc in structural domains, they do transitorily bind zinc at allosteric sites, modifying transiently the receptor channel's ion permeability. The present contribution summarizes current information showing that zinc allosteric modulation of receptor channels occurs by the preferential metal coordination to imidazole rings as well as to the sulfhydryl groups of cysteine in addition to the carboxyl group of acid residues, as with enzymes and catalysis. It is remarkable that most channels, either voltage-sensitive or transmitter-gated receptor channels, are susceptible to zinc modulation either as positive or negative regulators.

  1. Zinc as Allosteric Ion Channel Modulator: Ionotropic Receptors as Metalloproteins

    PubMed Central

    Peralta, Francisco Andrés; Huidobro-Toro, Juan Pablo

    2016-01-01

    Zinc is an essential metal to life. This transition metal is a structural component of many proteins and is actively involved in the catalytic activity of cell enzymes. In either case, these zinc-containing proteins are metalloproteins. However, the amino acid residues that serve as ligands for metal coordination are not necessarily the same in structural proteins compared to enzymes. While crystals of structural proteins that bind zinc reveal a higher preference for cysteine sulfhydryls rather than histidine imidazole rings, catalytic enzymes reveal the opposite, i.e., a greater preference for the histidines over cysteines for catalysis, plus the influence of carboxylic acids. Based on this paradigm, we reviewed the putative ligands of zinc in ionotropic receptors, where zinc has been described as an allosteric modulator of channel receptors. Although these receptors do not strictly qualify as metalloproteins since they do not normally bind zinc in structural domains, they do transitorily bind zinc at allosteric sites, modifying transiently the receptor channel’s ion permeability. The present contribution summarizes current information showing that zinc allosteric modulation of receptor channels occurs by the preferential metal coordination to imidazole rings as well as to the sulfhydryl groups of cysteine in addition to the carboxyl group of acid residues, as with enzymes and catalysis. It is remarkable that most channels, either voltage-sensitive or transmitter-gated receptor channels, are susceptible to zinc modulation either as positive or negative regulators. PMID:27384555

  2. Use of binding enthalpy to drive an allosteric transition.

    PubMed

    Brown, Patrick H; Beckett, Dorothy

    2005-03-01

    The Escherichia coli biotin repressor is an allosteric DNA binding protein and is activated by the small molecule bio-5'-AMP. Binding of this small molecule promotes transcription repression complex assembly between the repressor and the biotin operator of the biotin biosynthetic operon. The ability of the adenylate to activate the assembly process reflects its effect on biotin repressor dimerization. Thus concomitant with small molecule binding the free energy of repressor dimerization becomes more favorable by approximately -4 kcal/mol. The structural, dynamic, and energetic changes in the repressor monomer that accompany allosteric activation are not known. In this work the thermodynamics of binding of four allosteric activators to the repressor have been characterized by isothermal titration calorimetry. While binding of two of the effectors results in relatively modest activation of the dimerization process, binding of the other two small molecules, including the physiological effector, leads to large changes in repressor dimerization energetics. Results of the calorimetric measurements indicate that strong effector binding is accompanied by an enthalpically costly transition in the protein. This transition is "paid for" by the enthalpy that would have otherwise been realized from the formation of noncovalent bonds between the ligand and repressor monomer.

  3. Dynamics of allosteric transitions in GroEL

    PubMed Central

    Hyeon, Changbong; Lorimer, George H.; Thirumalai, D.

    2006-01-01

    The chaperonin GroEL-GroES, a machine that helps proteins to fold, cycles through a number of allosteric states, the T state, with high affinity for substrate proteins, the ATP-bound R state, and the R″ (GroEL–ADP–GroES) complex. Here, we use a self-organized polymer model for the GroEL allosteric states and a general structure-based technique to simulate the dynamics of allosteric transitions in two subunits of GroEL and the heptamer. The T → R transition, in which the apical domains undergo counterclockwise motion, is mediated by a multiple salt-bridge switch mechanism, in which a series of salt-bridges break and form. The initial event in the R → R″ transition, during which GroEL rotates clockwise, involves a spectacular outside-in movement of helices K and L that results in K80-D359 salt-bridge formation. In both the transitions there is considerable heterogeneity in the transition pathways. The transition state ensembles (TSEs) connecting the T, R, and R″ states are broad with the TSE for the T → R transition being more plastic than the R → R″ TSE. PMID:17135353

  4. Identification of the Allosteric Regulatory Site of Insulysin

    SciTech Connect

    Noinaj, Nicholas; Bhasin, Sonia K.; Song, Eun Suk; Scoggin, Kirsten E.; Juliano, Maria A.; Juliano, Luiz; Hersh, Louis B.; Rodgers, David W.

    2012-05-25

    Insulin degrading enzyme (IDE) is responsible for the metabolism of insulin and plays a role in clearance of the A{beta} peptide associated with Alzheimer's disease. Unlike most proteolytic enzymes, IDE, which consists of four structurally related domains and exists primarily as a dimer, exhibits allosteric kinetics, being activated by both small substrate peptides and polyphosphates such as ATP. The crystal structure of a catalytically compromised mutant of IDE has electron density for peptide ligands bound at the active site in domain 1 and a distal site in domain 2. Mutating residues in the distal site eliminates allosteric kinetics and activation by a small peptide, as well as greatly reducing activation by ATP, demonstrating that this site plays a key role in allostery. Comparison of the peptide bound IDE structure (using a low activity E111F IDE mutant) with unliganded wild type IDE shows a change in the interface between two halves of the clamshell-like molecule, which may enhance enzyme activity by altering the equilibrium between closed and open conformations. In addition, changes in the dimer interface suggest a basis for communication between subunits. Our findings indicate that a region remote from the active site mediates allosteric activation of insulysin by peptides. Activation may involve a small conformational change that weakens the interface between two halves of the enzyme.

  5. Identification of the Allosteric Regulatory Site of Insulysin

    SciTech Connect

    Noinaj, Nicholas; Bhasin, Sonia K.; Song, Eun Suk; Scoggin, Kirsten E.; Juliano, Maria A.; Juliano, Luiz; Hersh, Louis B.; Rodgers, David W.; Gerrard, Juliet Ann

    2011-06-24

    Background Insulin degrading enzyme (IDE) is responsible for the metabolism of insulin and plays a role in clearance of the Aβ peptide associated with Alzheimer's disease. Unlike most proteolytic enzymes, IDE, which consists of four structurally related domains and exists primarily as a dimer, exhibits allosteric kinetics, being activated by both small substrate peptides and polyphosphates such as ATP. Principal Findings The crystal structure of a catalytically compromised mutant of IDE has electron density for peptide ligands bound at the active site in domain 1 and a distal site in domain 2. Mutating residues in the distal site eliminates allosteric kinetics and activation by a small peptide, as well as greatly reducing activation by ATP, demonstrating that this site plays a key role in allostery. Comparison of the peptide bound IDE structure (using a low activity E111F IDE mutant) with unliganded wild type IDE shows a change in the interface between two halves of the clamshell-like molecule, which may enhance enzyme activity by altering the equilibrium between closed and open conformations. In addition, changes in the dimer interface suggest a basis for communication between subunits. Conclusions/Significance Our findings indicate that a region remote from the active site mediates allosteric activation of insulysin by peptides. Activation may involve a small conformational change that weakens the interface between two halves of the enzyme.

  6. NbIT - A New Information Theory-Based Analysis of Allosteric Mechanisms Reveals Residues that Underlie Function in the Leucine Transporter LeuT

    PubMed Central

    LeVine, Michael V.; Weinstein, Harel

    2014-01-01

    Complex networks of interacting residues and microdomains in the structures of biomolecular systems underlie the reliable propagation of information from an input signal, such as the concentration of a ligand, to sites that generate the appropriate output signal, such as enzymatic activity. This information transduction often carries the signal across relatively large distances at the molecular scale in a form of allostery that is essential for the physiological functions performed by biomolecules. While allosteric behaviors have been documented from experiments and computation, the mechanism of this form of allostery proved difficult to identify at the molecular level. Here, we introduce a novel analysis framework, called N-body Information Theory (NbIT) analysis, which is based on information theory and uses measures of configurational entropy in a biomolecular system to identify microdomains and individual residues that act as (i)-channels for long-distance information sharing between functional sites, and (ii)-coordinators that organize dynamics within functional sites. Application of the new method to molecular dynamics (MD) trajectories of the occluded state of the bacterial leucine transporter LeuT identifies a channel of allosteric coupling between the functionally important intracellular gate and the substrate binding sites known to modulate it. NbIT analysis is shown also to differentiate residues involved primarily in stabilizing the functional sites, from those that contribute to allosteric couplings between sites. NbIT analysis of MD data thus reveals rigorous mechanistic elements of allostery underlying the dynamics of biomolecular systems. PMID:24785005

  7. NbIT--a new information theory-based analysis of allosteric mechanisms reveals residues that underlie function in the leucine transporter LeuT.

    PubMed

    LeVine, Michael V; Weinstein, Harel

    2014-05-01

    Complex networks of interacting residues and microdomains in the structures of biomolecular systems underlie the reliable propagation of information from an input signal, such as the concentration of a ligand, to sites that generate the appropriate output signal, such as enzymatic activity. This information transduction often carries the signal across relatively large distances at the molecular scale in a form of allostery that is essential for the physiological functions performed by biomolecules. While allosteric behaviors have been documented from experiments and computation, the mechanism of this form of allostery proved difficult to identify at the molecular level. Here, we introduce a novel analysis framework, called N-body Information Theory (NbIT) analysis, which is based on information theory and uses measures of configurational entropy in a biomolecular system to identify microdomains and individual residues that act as (i)-channels for long-distance information sharing between functional sites, and (ii)-coordinators that organize dynamics within functional sites. Application of the new method to molecular dynamics (MD) trajectories of the occluded state of the bacterial leucine transporter LeuT identifies a channel of allosteric coupling between the functionally important intracellular gate and the substrate binding sites known to modulate it. NbIT analysis is shown also to differentiate residues involved primarily in stabilizing the functional sites, from those that contribute to allosteric couplings between sites. NbIT analysis of MD data thus reveals rigorous mechanistic elements of allostery underlying the dynamics of biomolecular systems.

  8. Prediction of allosteric sites on protein surfaces with an elastic-network-model-based thermodynamic method

    NASA Astrophysics Data System (ADS)

    Su, Ji Guo; Qi, Li Sheng; Li, Chun Hua; Zhu, Yan Ying; Du, Hui Jing; Hou, Yan Xue; Hao, Rui; Wang, Ji Hua

    2014-08-01

    Allostery is a rapid and efficient way in many biological processes to regulate protein functions, where binding of an effector at the allosteric site alters the activity and function at a distant active site. Allosteric regulation of protein biological functions provides a promising strategy for novel drug design. However, how to effectively identify the allosteric sites remains one of the major challenges for allosteric drug design. In the present work, a thermodynamic method based on the elastic network model was proposed to predict the allosteric sites on the protein surface. In our method, the thermodynamic coupling between the allosteric and active sites was considered, and then the allosteric sites were identified as those where the binding of an effector molecule induces a large change in the binding free energy of the protein with its ligand. Using the proposed method, two proteins, i.e., the 70 kD heat shock protein (Hsp70) and GluA2 alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor, were studied and the allosteric sites on the protein surface were successfully identified. The predicted results are consistent with the available experimental data, which indicates that our method is a simple yet effective approach for the identification of allosteric sites on proteins.

  9. Implementation of orthogonal frequency division multiplexing (OFDM) and advanced signal processing for elastic optical networking in accordance with networking and transmission constraints

    NASA Astrophysics Data System (ADS)

    Johnson, Stanley

    An increasing adoption of digital signal processing (DSP) in optical fiber telecommunication has brought to the fore several interesting DSP enabled modulation formats. One such format is orthogonal frequency division multiplexing (OFDM), which has seen great success in wireless and wired RF applications, and is being actively investigated by several research groups for use in optical fiber telecom. In this dissertation, I present three implementations of OFDM for elastic optical networking and distributed network control. The first is a field programmable gate array (FPGA) based real-time implementation of a version of OFDM conventionally known as intensity modulation and direct detection (IMDD) OFDM. I experimentally demonstrate the ability of this transmission system to dynamically adjust bandwidth and modulation format to meet networking constraints in an automated manner. To the best of my knowledge, this is the first real-time software defined networking (SDN) based control of an OFDM system. In the second OFDM implementation, I experimentally demonstrate a novel OFDM transmission scheme that supports both direct detection and coherent detection receivers simultaneously using the same OFDM transmitter. This interchangeable receiver solution enables a trade-off between bit rate and equipment cost in network deployment and upgrades. I show that the proposed transmission scheme can provide a receiver sensitivity improvement of up to 1.73 dB as compared to IMDD OFDM. I also present two novel polarization analyzer based detection schemes, and study their performance using experiment and simulation. In the third implementation, I present an OFDM pilot-tone based scheme for distributed network control. The first instance of an SDN-based OFDM elastic optical network with pilot-tone assisted distributed control is demonstrated. An improvement in spectral efficiency and a fast reconfiguration time of 30 ms have been achieved in this experiment. Finally, I

  10. Extracellular Calcium Modulates Actions of Orthosteric and Allosteric Ligands on Metabotropic Glutamate Receptor 1α*

    PubMed Central

    Jiang, Jason Y.; Nagaraju, Mulpuri; Meyer, Rebecca C.; Zhang, Li; Hamelberg, Donald; Hall, Randy A.; Brown, Edward M.; Conn, P. Jeffrey; Yang, Jenny J.

    2014-01-01

    Metabotropic glutamate receptor 1α (mGluR1α), a member of the family C G protein-coupled receptors, is emerging as a potential drug target for various disorders, including chronic neuronal degenerative diseases. In addition to being activated by glutamate, mGluR1α is also modulated by extracellular Ca2+. However, the underlying mechanism is unknown. Moreover, it has long been challenging to develop receptor-specific agonists due to homologies within the mGluR family, and the Ca2+-binding site(s) on mGluR1α may provide an opportunity for receptor-selective targeting by therapeutics. In the present study, we show that our previously predicted Ca2+-binding site in the hinge region of mGluR1α is adjacent to the site where orthosteric agonists and antagonists bind on the extracellular domain of the receptor. Moreover, we found that extracellular Ca2+ enhanced mGluR1α-mediated intracellular Ca2+ responses evoked by the orthosteric agonist l-quisqualate. Conversely, extracellular Ca2+ diminished the inhibitory effect of the mGluR1α orthosteric antagonist (S)-α-methyl-4-carboxyphenylglycine. In addition, selective positive (Ro 67-4853) and negative (7-(hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester) allosteric modulators of mGluR1α potentiated and inhibited responses to extracellular Ca2+, respectively, in a manner similar to their effects on the response of mGluR1α to glutamate. Mutations at residues predicted to be involved in Ca2+ binding, including E325I, had significant effects on the modulation of responses to the orthosteric agonist l-quisqualate and the allosteric modulator Ro 67-4853 by extracellular Ca2+. These studies reveal that binding of extracellular Ca2+ to the predicted Ca2+-binding site in the extracellular domain of mGluR1α modulates not only glutamate-evoked signaling but also the actions of both orthosteric ligands and allosteric modulators on mGluR1α. PMID:24280223

  11. Illumination distribution and signal transmission for indoor visible light communication with different light-emitting diode arrays and pre-equality circuits

    NASA Astrophysics Data System (ADS)

    Chen, Hsi-Chao; Liou, Cheng-Jyun; Siao, Syuan-Ruei

    2015-11-01

    The purpose of this study was to seek the optimal design for light-emitting diode (LED) arrays and pre-equality circuits in indoor visible lighting illumination combined with communication. The optical and communicational properties of illumination distribution and signal transmission were investigated. These illumination distributions of array sources were derivate and simulated and actually can be used in free-space communication. Simulated results show the total flux size was rectangle>radial>circlearray, and real measurements also showed the total flux was rectangle>radial>circlearray. The simulated and measured results have a similarity of over 98% by normalized cross correlation. In addition, when the distance of the installed lamp from the wall was 1 m, the rectangular array had the best illumination uniformity of 77.24%, and the size of uniformity was the rectangle>radial≈circle array. Finally, the gain and constant-current pre-equality circuits were used in free-space communication with a carrier frequency from 1 KHz to 1 MHz at a distance of 1.8 m. Both the received signal intensity and divergence angle were rectangle>radial>circle array. The constant-current pre-equality circuit could add the divergence angle from ±18.6 deg to ±36.68 deg in the rectangle array at a carrier frequency of 1 MHz.

  12. Reviews Book: SEP Communications: Transmitting and Receiving Signals Book: Gliding for Gold Book: Radioactivity: A History of a Mysterious Science Book: The New Quantum Age Books: The Art of Science and The Oxford Book of Modern Science Writing Equipment: SEP Analogue/digital transmission unit Equipment: SEP Optical signal transmission set Book: Stars and their Spectra Book: Voicebox: The Physics and Evolution of Speech Web Watch

    NASA Astrophysics Data System (ADS)

    2012-03-01

    WE RECOMMEND Transmitting and Receiving Signals SEP booklet transmits knowledge The New Quantum Age Understanding modern quantum theory The Art of Science and The Oxford Book of Modern Science Writing Anthologies bring science to life SEP Analogue/digital transmission unit Kit transmits signal between two points SEP Optical signal transmission set Optical kit shows light transmission Stars and their Spectra New book for teaching astrophysics WORTH A LOOK Gliding for Gold Take a journey through the physics of winter sports Radioactivity: A History of a Mysterious Science Book looks at history of radioactivity Voicebox: The Physics and Evolution of Speech TExploring the evolution of the voice WEB WATCH An interactive program with promise?

  13. Robust Stimulation of W1282X-CFTR Channel Activity by a Combination of Allosteric Modulators

    PubMed Central

    Wang, Wei; Hong, Jeong S.; Rab, Andras; Sorscher, Eric J.; Kirk, Kevin L.

    2016-01-01

    W1282X is a common nonsense mutation among cystic fibrosis patients that results in the production of a truncated Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel. Here we show that the channel activity of the W1282X-CFTR polypeptide is exceptionally low in excised membrane patches at normally saturating doses of ATP and PKA (single channel open probability (PO) < 0.01). However, W1282X-CFTR channels were stimulated by two CFTR modulators, the FDA-approved VX-770 and the dietary compound curcumin. Each of these compounds is an allosteric modulator of CFTR gating that promotes channel activity in the absence of the native ligand, ATP. Although W1282X-CFTR channels were stimulated by VX-770 in the absence of ATP their activities remained dependent on PKA phosphorylation. Thus, activated W1282X-CFTR channels should remain under physiologic control by cyclic nucleotide signaling pathways in vivo. VX-770 and curcumin exerted additive effects on W1282X-CFTR channel gating (opening/closing) in excised patches such that the Po of the truncated channel approached unity (> 0.9) when treated with both modulators. VX-770 and curcumin also additively stimulated W1282X-CFTR mediated currents in polarized FRT epithelial monolayers. In this setting, however, the stimulated W1282X-CFTR currents were smaller than those mediated by wild type CFTR (3–5%) due presumably to lower expression levels or cell surface targeting of the truncated protein. Combining allosteric modulators of different mechanistic classes is worth considering as a treatment option for W1282X CF patients perhaps when coupled with maneuvers to increase expression of the truncated protein. PMID:27007499

  14. Insights into the allosteric regulation of Syk association with receptor ITAM, a multi-state equilibrium.

    PubMed

    Feng, Chao; Post, Carol Beth

    2016-02-17

    The phosphorylation of interdomain A (IA), a linker region between tandem SH2 domains of Syk tyrosine kinase, regulates the binding affinity for association of Syk with doubly-phosphorylated ITAM regions of the B cell receptor. The mechanism of this allosteric regulation has been suggested to be a switch from the high-affinity bifunctional binding, mediated through both SH2 domains binding two phosphotyrosine residues of ITAM, to a substantially lower-affinity binding of only one SH2 domain. IA phosphorylation triggers the switch by inducing disorder in IA and weakening the SH2-SH2 interaction. The postulated switch to a single-SH2-domain binding mode is examined using NMR to monitor site-specific binding to each SH2 domain of Syk variants engineered to have IA regions that differ in conformational flexibility. The combined analysis of titration curves and NMR line-shapes provides sufficient information to determine the energetics of inter-molecular binding at each SH2 site along with an intra-molecular binding or isomerization step. A less favorable isomerization equilibrium associated with the changes in the SH2-SH2 conformational ensemble and IA flexibility accounts for the inhibition of Syk association with membrane ITAM regions when IA is phosphorylated, and refutes the proposed switch to single-SH2-domain binding. Syk localizes in the cell through its SH2 interactions, and this basis for allosteric regulation of ITAM association proposes for the first time a phosphorylation-dependent model to regulate Syk binding to alternate receptors and other signaling proteins that differ either in the number of residues separating ITAM phosphotyrosines or by having only one phosphotyrosine, a half ITAM.

  15. Robust Stimulation of W1282X-CFTR Channel Activity by a Combination of Allosteric Modulators.

    PubMed

    Wang, Wei; Hong, Jeong S; Rab, Andras; Sorscher, Eric J; Kirk, Kevin L

    2016-01-01

    W1282X is a common nonsense mutation among cystic fibrosis patients that results in the production of a truncated Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) channel. Here we show that the channel activity of the W1282X-CFTR polypeptide is exceptionally low in excised membrane patches at normally saturating doses of ATP and PKA (single channel open probability (PO) < 0.01). However, W1282X-CFTR channels were stimulated by two CFTR modulators, the FDA-approved VX-770 and the dietary compound curcumin. Each of these compounds is an allosteric modulator of CFTR gating that promotes channel activity in the absence of the native ligand, ATP. Although W1282X-CFTR channels were stimulated by VX-770 in the absence of ATP their activities remained dependent on PKA phosphorylation. Thus, activated W1282X-CFTR channels should remain under physiologic control by cyclic nucleotide signaling pathways in vivo. VX-770 and curcumin exerted additive effects on W1282X-CFTR channel gating (opening/closing) in excised patches such that the Po of the truncated channel approached unity (> 0.9) when treated with both modulators. VX-770 and curcumin also additively stimulated W1282X-CFTR mediated currents in polarized FRT epithelial monolayers. In this setting, however, the stimulated W1282X-CFTR currents were smaller than those mediated by wild type CFTR (3-5%) due presumably to lower expression levels or cell surface targeting of the truncated protein. Combining allosteric modulators of different mechanistic classes is worth considering as a treatment option for W1282X CF patients perhaps when coupled with maneuvers to increase expression of the truncated protein.

  16. Allosteric Inhibition of the Neuropeptidase Neurolysin*

    PubMed Central

    Hines, Christina S.; Ray, Kallol; Schmidt, Jack J.; Xiong, Fei; Feenstra, Rolf W.; Pras-Raves, Mia; de Moes, Jan Peter; Lange, Jos H. M.; Melikishvili, Manana; Fried, Michael G.; Mortenson, Paul; Charlton, Michael; Patel, Yogendra; Courtney, Stephen M.; Kruse, Chris G.; Rodgers, David W.

    2014-01-01

    Neuropeptidases specialize in the hydrolysis of the small bioactive peptides that play a variety of signaling roles in the nervous and endocrine systems. One neuropeptidase, neurolysin, helps control the levels of the dopaminergic circuit modulator neurotensin and is a member of a fold group that includes the antihypertensive target angiotensin converting enzyme. We report the discovery of a potent inhibitor that, unexpectedly, binds away from the enzyme catalytic site. The location of the bound inhibitor suggests it disrupts activity by preventing a hinge-like motion associated with substrate binding and catalysis. In support of this model, the inhibition kinetics are mixed, with both noncompetitive and competitive components, and fluorescence polarization shows directly that the inhibitor reverses a substrate-associated conformational change. This new type of inhibition may have widespread utility in targeting neuropeptidases. PMID:25378390

  17. Allosteric inhibition of the neuropeptidase neurolysin.

    PubMed

    Hines, Christina S; Ray, Kallol; Schmidt, Jack J; Xiong, Fei; Feenstra, Rolf W; Pras-Raves, Mia; de Moes, Jan Peter; Lange, Jos H M; Melikishvili, Manana; Fried, Michael G; Mortenson, Paul; Charlton, Michael; Patel, Yogendra; Courtney, Stephen M; Kruse, Chris G; Rodgers, David W

    2014-12-19

    Neuropeptidases specialize in the hydrolysis of the small bioactive peptides that play a variety of signaling roles in the nervous and endocrine systems. One neuropeptidase, neurolysin, helps control the levels of the dopaminergic circuit modulator neurotensin and is a member of a fold group that includes the antihypertensive target angiotensin converting enzyme. We report the discovery of a potent inhibitor that, unexpectedly, binds away from the enzyme catalytic site. The location of the bound inhibitor suggests it disrupts activity by preventing a hinge-like motion associated with substrate binding and catalysis. In support of this model, the inhibition kinetics are mixed, with both noncompetitive and competitive components, and fluorescence polarization shows directly that the inhibitor reverses a substrate-associated conformational change. This new type of inhibition may have widespread utility in targeting neuropeptidases.

  18. Efficacy of selective PDE4D negative allosteric modulators in the object retrieval task in female cynomolgus monkeys (Macaca fascicularis).

    PubMed

    Sutcliffe, Jane S; Beaumont, Vahri; Watson, James M; Chew, Chang Sing; Beconi, Maria; Hutcheson, Daniel M; Dominguez, Celia; Munoz-Sanjuan, Ignacio

    2014-01-01

    Cyclic adenosine monophosphate (cAMP) signalling plays an important role in synaptic plasticity and information processing in the hippocampal and basal ganglia systems. The augmentation of cAMP signalling through the selective inhibition of phosphodiesterases represents a viable strategy to treat disorders associated with dysfunction of these circuits. The phosphodiesterase (PDE) type 4 inhibitor rolipram has shown significant pro-cognitive effects in neurological disease models, both in rodents and primates. However, competitive non-isoform selective PDE4 inhibitors have a low therapeutic index which has stalled their clinical development. Here, we demonstrate the pro-cognitive effects of selective negative allosteric modulators (NAMs) of PDE4D, D159687 and D159797 in female Cynomolgous macaques, in the object retrieval detour task. The efficacy displayed by these NAMs in a primate cognitive task which engages the corticostriatal circuitry, together with their suitable pharmacokinetic properties and safety profiles, suggests that clinical development of these allosteric modulators should be considered for the treatment of a variety of brain disorders associated with cognitive decline.

  19. Efficacy of Selective PDE4D Negative Allosteric Modulators in the Object Retrieval Task in Female Cynomolgus Monkeys (Macaca fascicularis)

    PubMed Central

    Sutcliffe, Jane S.; Beaumont, Vahri; Watson, James M.; Chew, Chang Sing; Beconi, Maria; Hutcheson, Daniel M.; Dominguez, Celia; Munoz-Sanjuan, Ignacio

    2014-01-01

    Cyclic adenosine monophosphate (cAMP) signalling plays an important role in synaptic plasticity and information processing in the hippocampal and basal ganglia systems. The augmentation of cAMP signalling through the selective inhibition of phosphodiesterases represents a viable strategy to treat disorders associated with dysfunction of these circuits. The phosphodiesterase (PDE) type 4 inhibitor rolipram has shown significant pro-cognitive effects in neurological disease models, both in rodents and primates. However, competitive non-isoform selective PDE4 inhibitors have a low therapeutic index which has stalled their clinical development. Here, we demonstrate the pro-cognitive effects of selective negative allosteric modulators (NAMs) of PDE4D, D159687 and D159797 in female Cynomolgous macaques, in the object retrieval detour task. The efficacy displayed by these NAMs in a primate cognitive task which engages the corticostriatal circuitry, together with their suitable pharmacokinetic properties and safety profiles, suggests that clinical development of these allosteric modulators should be considered for the treatment of a variety of brain disorders associated with cognitive decline. PMID:25050979

  20. Allosteric Inhibition of a Semaphorin 4D Receptor Plexin B1 by a High-Affinity Macrocyclic Peptide.

    PubMed

    Matsunaga, Yukiko; Bashiruddin, Nasir K; Kitago, Yu; Takagi, Junichi; Suga, Hiroaki

    2016-11-17

    Semaphorin axonal guidance factors are multifunctional proteins that play important roles in immune response, cancer cell proliferation, and organogenesis, making semaphorins and their signaling receptor plexins important drug targets for various diseases. However, the large and flat binding surface of the semaphorin-plexin interaction interface is difficult to target by traditional small-molecule drugs. Here, we report the discovery of a high-affinity plexin B1 (PlxnB1)-binding macrocyclic peptide, PB1m6 (KD = 3.5 nM). PB1m6 specifically inhibited the binding of physiological ligand semaphorin 4D (Sema4D) in vitro and completely suppressed Sema4D-induced cell collapse. Structural studies revealed that PB1m6 binds at a groove between the fifth and sixth blades of the sema domain in PlxnB1 distant from the Sema4D-binding site, indicating the non-competitive and allosteric nature of the inhibitory activity. The discovery of this novel allosteric site can potentially be used to target plexin family proteins for the development of drugs that modulate semaphorin and plexin signaling.

  1. A negative allosteric modulator demonstrates biased antagonism of the follicle stimulating hormone receptor

    PubMed Central

    Dias, James A.; Bonnet, Béatrice; Weaver, Barbara A.; Watts, Julie; Kluetzman, Kerri; Thomas, Richard M.; Poli, Sonia; Mutel, Vincent; Campo, Brice

    2015-01-01

    High quality gamete production in males and females requires the pituitary gonadotropin follicle stimulating hormone (FSH). In this report a novel chemical class of small molecule inhibitors of FSH receptor (FSHR) is described. ADX61623, a negative allosteric modulator (NAM), increased the affinity of interaction between 125I-hFSH and human FSHR (hFSHR) five fold. This form of FSHR occupied simultaneously by FSH and ADX61623 was inactive for cAMP and progesterone production in primary cultures of rat granulosa cells. In contrast, ADX61623 did not block estrogen production. This demonstrates for the first time, biased antagonism at the FSHR. To determine if ADX61623 blocked FSH induction of follicle development in vivo, a bioassay to measure follicular development and oocyte production in immature female rats was validated. ADX61623 was not completely effective in blocking FSH induced follicular development in vivo at doses up to 100 mg/kg as oocyte production and ovarian weight gain were only moderately reduced. These data illustrate that FSHR couples to multiple signaling pathways in vivo. Suppression of one pool of FSHR uncouples Gαs and cAMP production, and decreases progesterone production. Occupancy of another pool of FSHR sensitizes granulosa cells to FSH induced estradiol production. Therefore, ADX61623 is a useful tool to investigate further the mechanism of the FSHR signaling dichotomy. This may lead to a greater understanding of the signaling infrastructure which enables estrogen biosynthesis and may prove useful in treating estrogen dependent disease. PMID:21184806

  2. Discovery of multiple hidden allosteric sites by combining Markov state models and experiments.

    PubMed

    Bowman, Gregory R; Bolin, Eric R; Hart, Kathryn M; Maguire, Brendan C; Marqusee, Susan

    2015-03-03

    The discovery of drug-like molecules that bind pockets in proteins that are not present in crystallographic structures yet exert allosteric control over activity has generated great interest in designing pharmaceuticals that exploit allosteric effects. However, there have only been a small number of successes, so the therapeutic potential of these pockets--called hidden allosteric sites--remains unclear. One challenge for assessing their utility is that rational drug design approaches require foreknowledge of the target site, but most hidden allosteric sites are only discovered when a small molecule is found to stabilize them. We present a means of decoupling the identification of hidden allosteric sites from the discovery of drugs that bind them by drawing on new developments in Markov state modeling that provide unprecedented access to microsecond- to millisecond-timescale fluctuations of a protein's structure. Visualizing these fluctuations allows us to identify potential hidden allosteric sites, which we then test via thiol labeling experiments. Application of these methods reveals multiple hidden allosteric sites in an important antibiotic target--TEM-1 β-lactamase. This result supports the hypothesis that there are many as yet undiscovered hidden allosteric sites and suggests our methodology can identify such sites, providing a starting point for future drug design efforts. More generally, our results demonstrate the power of using Markov state models to guide experiments.

  3. Allosteric regulation of glycogen synthase in liver. A physiological dilemma.

    PubMed

    Nuttall, F Q; Gannon, M C

    1993-06-25

    Glycogen synthase catalyzes the transfer of the glucosyl moiety from UDP-glucose to the terminal branch of the glycogen molecule and is considered to be the rate-limiting enzyme for glycogen synthesis. However, under ideal assay conditions, i.e. 37 degrees C with saturating concentrations of UDP-glucose and the activator, glucose-6-P, the maximal catalytic activity of glycogen synthase was only 78% of the in vivo glycogen synthetic rate. Using concentrations of UDP-glucose and glucose-6-P likely to be present in vivo, the rate was only approximately 30%. This prompted us to reassess a possible role of allosteric effectors on synthase activity. Glycogen synthase was assayed at 37 degrees C using dilute, pH 7.0, buffered extracts, initial rate conditions, and UDP-glucose and glucose-6-P concentrations, which approximate those calculated to be present in total liver cell water. Several allosteric effectors were tested. Magnesium and AMP had little effect on activity. Pi, ADP, ATP, and UTP inhibited activity. When a combination of effectors were added at concentrations approximating those present in cell water, synthase activity could account for only 2% of the glycogen synthetic rate. Thus, although allosteric effectors are likely to be playing a major role in regulating synthase enzymic activity in liver cells, to date, a metabolite that can stimulate activity and/or overcome nucleotide inhibition has yet to be identified. If such a metabolite cannot be identified, an additional or alternative pathway for glycogen synthesis must be considered.

  4. Automated manual transmission controller

    DOEpatents

    Lawrie, Robert E.; Reed, Jr., Richard G.; Bernier, David R.

    1999-12-28

    A powertrain system for a hybrid vehicle. The hybrid vehicle includes a heat engine, such as a diesel engine, and an electric machine, which operates as both an electric motor and an alternator, to power the vehicle. The hybrid vehicle also includes a manual-style transmission configured to operate as an automatic transmission from the perspective of the driver. The engine and the electric machine drive an input shaft which in turn drives an output shaft of the transmission. In addition to driving the transmission, the electric machine regulates the speed of the input shaft in order to synchronize the input shaft during either an upshift or downshift of the transmission by either decreasing or increasing the speed of the input shaft. When decreasing the speed of the input shaft, the electric motor functions as an alternator to produce electrical energy which may be stored by a storage device. Operation of the transmission is controlled by a transmission controller which receives input signals and generates output signals to control shift and clutch motors to effect smooth launch, upshift shifts, and downshifts of the transmission, so that the transmission functions substantially as an automatic transmission from the perspective of the driver, while internally substantially functioning as a manual transmission.

  5. A novel allosteric mechanism in the cysteine peptidase cathepsin K discovered by computational methods

    NASA Astrophysics Data System (ADS)

    Novinec, Marko; Korenč, Matevž; Caflisch, Amedeo; Ranganathan, Rama; Lenarčič, Brigita; Baici, Antonio

    2014-02-01

    Allosteric modifiers have the potential to fine-tune enzyme activity. Therefore, targeting allosteric sites is gaining increasing recognition as a strategy in drug design. Here we report the use of computational methods for the discovery of the first small-molecule allosteric inhibitor of the collagenolytic cysteine peptidase cathepsin K, a major target for the treatment of osteoporosis. The molecule NSC13345 is identified by high-throughput docking of compound libraries to surface sites on the peptidase that are connected to the active site by an evolutionarily conserved network of residues (protein sector). The crystal structure of the complex shows that NSC13345 binds to a novel allosteric site on cathepsin K. The compound acts as a hyperbolic mixed modifier in the presence of a synthetic substrate, it completely inhibits collagen degradation and has good selectivity for cathepsin K over related enzymes. Altogether, these properties qualify our methodology and NSC13345 as promising candidates for allosteric drug design.

  6. Experimental evidence for allosteric modifier saturation as predicted by the bi-substrate Hill equation.

    PubMed

    Hanekom, A J; Hofmeyr, J H S; Snoep, J L; Rohwer, J M

    2006-09-01

    The cooperative enzyme reaction rates predicted by the bi-substrate Hill equation and the bi-substrate Monod-Wyman-Changeux (MWC) equation when allosterically inhibited are compared in silico. Theoretically, the Hill equation predicts that when the maximum inhibitory effect at a certain substrate condition has been reached, an increase in allosteric inhibitor concentration will have no effect on reaction rate, that is the Hill equation shows allosteric inhibitor saturation. This saturating inhibitory effect is not present in the MWC equation. Experimental in vitro data for pyruvate kinase, a bi-substrate cooperative enzyme that is allosterically inhibited, are presented. This enzyme also shows inhibitor saturation, and therefore serves as experimental evidence that the bi-substrate Hill equation predicts more realistic allosteric inhibitor behaviour than the bi-substrate MWC equation.

  7. Bioinformatic scaling of allosteric interactions in biomedical isozymes

    NASA Astrophysics Data System (ADS)

    Phillips, J. C.

    2016-09-01

    Allosteric (long-range) interactions can be surprisingly strong in proteins of biomedical interest. Here we use bioinformatic scaling to connect prior results on nonsteroidal anti-inflammatory drugs to promising new drugs that inhibit cancer cell metabolism. Many parallel features are apparent, which explain how even one amino acid mutation, remote from active sites, can alter medical results. The enzyme twins involved are cyclooxygenase (aspirin) and isocitrate dehydrogenase (IDH). The IDH results are accurate to 1% and are overdetermined by adjusting a single bioinformatic scaling parameter. It appears that the final stage in optimizing protein functionality may involve leveling of the hydrophobic limits of the arms of conformational hydrophilic hinges.

  8. Transmission of single-carrier 400G signals (515.2-Gb/s) based on 128.8-GBaud PDM QPSK over 10,130- and 6,078 km terrestrial fiber links.

    PubMed

    Zhang, Junwen; Yu, Jianjun; Zhu, Benyuan; Li, Fan; Chien, Hung-Chang; Jia, Zhensheng; Cai, Yi; Li, Xinying; Xiao, Xin; Fang, Yuan; Wang, Yuanquan

    2015-06-29

    We experimentally demonstrate the coherent transmission system with the highest ETDM-based symbol rate of 128.8-GBaud over record breaking distances. We successfully transmitted single-carrier 515.2-Gb/s PDM-QPSK/9-QAM signals over 10,130km/6,078-km, respectively, over 100km spans of TeraWave SLA + fiber. To the best of our knowledge, it is the highest ETDM-based symbol rate reported so far, and the longest WDM transmission distance with single-carrier 400G signals. For the first time, the 515.2-Gb/s single-carrier PDM-QPSK signals in 200-GHz-grid are successfully transmitted over distance above 10,000km in terrestrial transmission environment. We have also demonstrated the transmission of single carrier 128.8-GBaud filtered QPSK signals in 100-GHz-grid over 6,078-km, which has the line spectral efficiency (SE) of 5.152 (b/s/Hz).

  9. Allosteric modulators of the hERG K{sup +} channel

    SciTech Connect

    Yu, Zhiyi Klaasse, Elisabeth Heitman, Laura H. IJzerman, Adriaan P.

    2014-01-01

    Drugs that block the cardiac K{sup +} channel encoded by the human ether-à-go-go gene (hERG) have been associated with QT interval prolongation leading to proarrhythmia, and in some cases, sudden cardiac death. Because of special structural features of the hERG K{sup +} channel, it has become a promiscuous target that interacts with pharmaceuticals of widely varying chemical structures and a reason for concern in the pharmaceutical industry. The structural diversity suggests that multiple binding sites are available on the channel with possible allosteric interactions between them. In the present study, three reference compounds and nine compounds of a previously disclosed series were evaluated for their allosteric effects on the binding of [{sup 3}H]astemizole and [{sup 3}H]dofetilide to the hERG K{sup +} channel. LUF6200 was identified as an allosteric inhibitor in dissociation assays with both radioligands, yielding similar EC{sub 50} values in the low micromolar range. However, potassium ions increased the binding of the two radioligands in a concentration-dependent manner, and their EC{sub 50} values were not significantly different, indicating that potassium ions behaved as allosteric enhancers. Furthermore, addition of potassium ions resulted in a concentration-dependent leftward shift of the LUF6200 response curve, suggesting positive cooperativity and distinct allosteric sites for them. In conclusion, our investigations provide evidence for allosteric modulation of the hERG K{sup +} channel, which is discussed in the light of findings on other ion channels. - Highlights: • Allosteric modulators on the hERG K{sup +} channel were evaluated in binding assays. • LUF6200 was identified as a potent allosteric inhibitor. • Potassium ions were found to behave as allosteric enhancers. • Positive cooperativity and distinct allosteric sites for them were proposed.

  10. Real-time optical wireless transmissions of digital TV signals using white InGaN LEDs grown with an asymmetric quantum barrier.

    PubMed

    Tsai, Chia-Lung; Chen, Yen-Jen

    2015-10-19

    The feasibility of using InGaN LEDs grown with asymmetric barrier layer (ABL) as transmitters in visible light communications is investigated experimentally. Compared with normal LEDs, the improvement in the spontaneous emission rate due to enhanced carrier localization and better uniformity of carrier distribution in ABL-containing MQWs leads to the fabricated LEDs can exhibit a 32.6% (@ 350 mA) increase in emission intensity and a 10.5% increase in modulation bandwidth. After eliminating the slow-responding phosphorescent components emitting from the phosphor-converted white LEDs, an open eye-diagram at 180 Mb/s is demonstrated over a distance of 100 cm in directed line-of-sight optical links. With the use of proposed LEDs, real-time transmissions of digital TV signals over a moderate distance (~100 cm) in free space is shown to be available in a 150 Mbit/s white LED-based optical link with conventional on-off keying modulation.

  11. Hepacivirus NS3/4A Proteases Interfere with MAVS Signaling in both Their Cognate Animal Hosts and Humans: Implications for Zoonotic Transmission.

    PubMed

    Anggakusuma; Brown, Richard J P; Banda, Dominic H; Todt, Daniel; Vieyres, Gabrielle; Steinmann, Eike; Pietschmann, Thomas

    2016-12-01

    Multiple novel members of the genus Hepacivirus have recently been discovered in diverse mammalian species. However, to date, their replication mechanisms and zoonotic potential have not been explored in detail. The NS3/4A serine protease of hepatitis C virus (HCV) is critical for cleavage of the viral polyprotein. It also cleaves the cellular innate immune adaptor MAVS, thus decreasing interferon (IFN) production and contributing to HCV persistence in the human host. To investigate the conservation of fundamental aspects of the hepaciviral life cycle, we explored if MAVS cleavage and suppression of innate immune signaling represent a common mechanism employed across different clades of the genus Hepacivirus to enhance viral replication. To estimate the zoonotic potential of these nonhuman hepaciviruses, we assessed if their NS3/4A proteases were capable of cleaving human MAVS. NS3/4A proteases of viruses infecting colobus monkeys, rodents, horses, and cows cleaved the MAVS proteins of their cognate hosts and interfered with the ability of MAVS to induce the IFN-β promoter. All NS3/4A proteases from nonhuman viruses readily cleaved human MAVS. Thus, NS3/4A-dependent cleavage of MAVS is a conserved replication strategy across multiple clades within the genus Hepacivirus Human MAVS is susceptible to cleavage by these nonhuman viral proteases, indicating that it does not pose a barrier for zoonotic transmission of these viruses to humans.

  12. Variations between the photosynthetic properties of elite and landrace Chinese rice cultivars revealed by simultaneous measurements of 820 nm transmission signal and chlorophyll a fluorescence induction.

    PubMed

    Hamdani, Saber; Qu, Mingnan; Xin, Chang-Peng; Li, Ming; Chu, Chengcai; Govindjee; Zhu, Xin-Guang

    2015-04-01

    The difference between the photosynthetic properties of elite and landrace Chinese rice cultivars was studied, using chlorophyll a fluorescence induction (mostly a monitor of Photosystem II activity) and I820 transmission signal (mostly a monitor of Photosystem I activity) to identify potential photosynthetic features differentiating these two groups, which show different degrees of artificial selection and grain yields. A higher fluorescence (related to PSII) IP rise phase and a lower P700(+) (related to PSI) accumulation were observed in the elite cultivars as compared to the landraces. Using these data, together with simulation data from a kinetic model of fluorescence induction, we show that the high IP rise phase and the low P700(+) accumulation can be a result of transient block on electron transfer and traffic jam on the electron acceptor side of PSI under a high [NADPH]/[NADP(+)] ratio. Considering that the ferredoxin NADP(+) reductase (FNR) transcript levels of XS134 (a representative elite cultivars) remains unaffected during the first few minutes of light/dark transition compared to Q4145 (a representative landrace cultivars), which shows a strong decline during the same time range, we propose that the FNR of elite cultivars may take more time to be inactivated in darkness. During this time the FNR enzyme can continue to reduce NADP(+) molecules, leading to initially high [NADPH]/[NADP(+)] ratio during OJIP transient. These data suggested a potential artificial selection of FNR during the breeding process of these examined elite rice cultivars.

  13. Allosteric modulation of the RNA polymerase catalytic reaction is an essential component of transcription control by rifamycins.

    PubMed

    Artsimovitch, Irina; Vassylyeva, Marina N; Svetlov, Dmitri; Svetlov, Vladimir; Perederina, Anna; Igarashi, Noriyuki; Matsugaki, Naohiro; Wakatsuki, Soichi; Tahirov, Tahir H; Vassylyev, Dmitry G

    2005-08-12

    Rifamycins, the clinically important antibiotics, target bacterial RNA polymerase (RNAP). A proposed mechanism in which rifamycins sterically block the extension of nascent RNA beyond three nucleotides does not alone explain why certain RNAP mutations confer resistance to some but not other rifamycins. Here we show that unlike rifampicin and rifapentin, and contradictory to the steric model, rifabutin inhibits formation of the first and second phosphodiester bonds. We report 2.5 A resolution structures of rifabutin and rifapentin complexed with the Thermus thermophilus RNAP holoenzyme. The structures reveal functionally important distinct interactions of antibiotics with the initiation sigma factor. Strikingly, both complexes lack the catalytic Mg2+ ion observed in the apo-holoenzyme, whereas an increase in Mg2+ concentration confers resistance to rifamycins. We propose that a rifamycin-induced signal is transmitted over approximately 19 A to the RNAP active site to slow down catalysis. Based on structural predictions, we designed enzyme substitutions that apparently interrupt this allosteric signal.

  14. Discovery and Characterization of Allosteric WNK Kinase Inhibitors.

    PubMed

    Yamada, Ken; Zhang, Ji-Hu; Xie, Xiaoling; Reinhardt, Juergen; Xie, Amy Qiongshu; LaSala, Daniel; Kohls, Darcy; Yowe, David; Burdick, Debra; Yoshisue, Hajime; Wakai, Hiromichi; Schmidt, Isabel; Gunawan, Jason; Yasoshima, Kayo; Yue, Q Kimberley; Kato, Mitsunori; Mogi, Muneto; Idamakanti, Neeraja; Kreder, Natasha; Drueckes, Peter; Pandey, Pramod; Kawanami, Toshio; Huang, Waanjeng; Yagi, Yukiko I; Deng, Zhan; Park, Hyi-Man

    2016-12-16

    Protein kinases are known for their highly conserved adenosine triphosphate (ATP)-binding site, rendering the discovery of selective inhibitors a major challenge. In theory, allosteric inhibitors can achieve high selectivity by targeting less conserved regions of the kinases, often with an added benefit of retaining efficacy under high physiological ATP concentration. Although often overlooked in favor of ATP-site directed approaches, performing a screen at high ATP concentration or stringent hit triaging with high ATP concentration offers conceptually simple methods of identifying inhibitors that bind outside the ATP pocket. Here, we applied the latter approach to the With-No-Lysine (K) (WNK) kinases to discover lead molecules for a next-generation antihypertensive that requires a stringent safety profile. This strategy yielded several ATP noncompetitive WNK1-4 kinase inhibitors, the optimization of which enabled cocrystallization with WNK1, revealing an allosteric binding mode consistent with the observed exquisite specificity for WNK1-4 kinases. The optimized compound inhibited rubidium uptake by sodium chloride cotransporter 1 (NKCC1) in HT29 cells, consistent with the reported physiology of WNK kinases in renal electrolyte handling.

  15. Structural Analysis of Iac Repressor Bound to Allosteric Effectors

    SciTech Connect

    Daber,R.; Stayrook, S.; Rosenberg, A.; Lewis, M.

    2007-01-01

    The lac operon is a model system for understanding how effector molecules regulate transcription and are necessary for allosteric transitions. The crystal structures of the lac repressor bound to inducer and anti-inducer molecules provide a model for how these small molecules can modulate repressor function. The structures of the apo repressor and the repressor bound to effector molecules are compared in atomic detail. All effectors examined here bind to the repressor in the same location and are anchored to the repressor through hydrogen bonds to several hydroxyl groups of the sugar ring. Inducer molecules form a more extensive hydrogen-bonding network compared to anti-inducers and neutral effector molecules. The structures of these effector molecules suggest that the O6 hydroxyl on the galactoside is essential for establishing a water-mediated hydrogen bonding network that bridges the N-terminal and C-terminal sub-domains. The altered hydrogen bonding can account in part for the different structural conformations of the repressor, and is vital for the allosteric transition.

  16. Targeting PARP-1 allosteric regulation offers therapeutic potential against cancer

    PubMed Central

    Steffen, Jamin D.; Tholey, Renee M.; Langelier, Marie-France; Planck, Jamie L.; Schiewer, Matthew J.; Lal, Shruti; Bildzukewicz, Nikolai A.; Yeo, Charles J.; Knudsen, Karen E.; Brody, Jonathan R.; Pascal, John M.

    2014-01-01

    PARP-1 is a nuclear protein that has important roles in maintenance of genomic integrity. During genotoxic stress, PARP-1 recruits to sites of DNA damage where PARP-1 domain architecture initiates catalytic activation and subsequent poly(ADP-ribose)-dependent DNA repair. PARP-1 inhibition is a promising new way to selectively target cancers harboring DNA repair deficiencies. However, current inhibitors target other PARPs raising important questions concerning long-term off-target effects. Here we propose a new strategy that targets PARP-1 allosteric regulation as a selective way of inhibiting PARP-1. We found that disruption of PARP-1 domain-domain contacts through mutagenesis held no cellular consequences on recruitment to DNA damage or a model system of transcriptional regulation, but prevented DNA-damage dependent catalytic activation. Further, PARP-1 mutant overexpression in a pancreatic cancer cell line (MIA PaCa-2) increased sensitivity to platinum-based anti-cancer agents. These results not only highlight the potential of a synergistic drug combination of allosteric PARP inhibitors with DNA damaging agents in genomically unstable cancer cells (regardless of homologous recombination status), but also signify important applications of selective PARP-1 inhibition. Lastly, the development of a high-throughput (HT) PARP-1 assay is described as a tool to promote discovery of novel PARP-1 selective inhibitors. PMID:24189460

  17. Targeting PARP-1 allosteric regulation offers therapeutic potential against cancer.

    PubMed

    Steffen, Jamin D; Tholey, Renee M; Langelier, Marie-France; Planck, Jamie L; Schiewer, Matthew J; Lal, Shruti; Bildzukewicz, Nikolai A; Yeo, Charles J; Knudsen, Karen E; Brody, Jonathan R; Pascal, John M

    2014-01-01

    PARP-1 is a nuclear protein that has important roles in maintenance of genomic integrity. During genotoxic stress, PARP-1 recruits to sites of DNA damage where PARP-1 domain architecture initiates catalytic activation and subsequent poly(ADP-ribose)-dependent DNA repair. PARP-1 inhibition is a promising new way to selectively target cancers harboring DNA repair deficiencies. However, current inhibitors target other PARPs, raising important questions about long-term off-target effects. Here, we propose a new strategy that targets PARP-1 allosteric regulation as a selective way of inhibiting PARP-1. We found that disruption of PARP-1 domain-domain contacts through mutagenesis held no cellular consequences on recruitment to DNA damage or a model system of transcriptional regulation, but prevented DNA-damage-dependent catalytic activation. Furthermore, PARP-1 mutant overexpression in a pancreatic cancer cell line (MIA PaCa-2) increased sensitivity to platinum-based anticancer agents. These results not only highlight the potential of a synergistic drug combination of allosteric PARP inhibitors with DNA-damaging agents in genomically unstable cancer cells (regardless of homologous recombination status), but also signify important applications of selective PARP-1 inhibition. Finally, the development of a high-throughput PARP-1 assay is described as a tool to promote discovery of novel PARP-1 selective inhibitors.

  18. The allosteric switching mechanism in bacteriophage MS2

    NASA Astrophysics Data System (ADS)

    Perkett, Matthew R.; Mirijanian, Dina T.; Hagan, Michael F.

    2016-07-01

    We use all-atom simulations to elucidate the mechanisms underlying conformational switching and allostery within the coat protein of the bacteriophage MS2. Assembly of most icosahedral virus capsids requires that the capsid protein adopts different conformations at precise locations within the capsid. It has been shown that a 19 nucleotide stem loop (TR) from the MS2 genome acts as an allosteric effector, guiding conformational switching of the coat protein during capsid assembly. Since the principal conformational changes occur far from the TR binding site, it is important to understand the molecular mechanism underlying this allosteric communication. To this end, we use all-atom simulations with explicit water combined with a path sampling technique to sample the MS2 coat protein conformational transition, in the presence and absence of TR-binding. The calculations find that TR binding strongly alters the transition free energy profile, leading to a switch in the favored conformation. We discuss changes in molecular interactions responsible for this shift. We then identify networks of amino acids with correlated motions to reveal the mechanism by which effects of TR binding span the protein. We find that TR binding strongly affects residues located at the 5-fold and quasi-sixfold interfaces in the assembled capsid, suggesting a mechanism by which the TR binding could direct formation of the native capsid geometry. The analysis predicts amino acids whose substitution by mutagenesis could alter populations of the conformational substates or their transition rates.

  19. A novel allosteric inhibitor of macrophage migration inhibitory factor (MIF).

    PubMed

    Bai, Fengwei; Asojo, Oluwatoyin A; Cirillo, Pier; Ciustea, Mihai; Ledizet, Michel; Aristoff, Paul A; Leng, Lin; Koski, Raymond A; Powell, Thomas J; Bucala, Richard; Anthony, Karen G

    2012-08-31

    Macrophage migration inhibitory factor (MIF) is a catalytic cytokine and an upstream mediator of the inflammatory pathway. MIF has broad regulatory properties, dysregulation of which has been implicated in the pathology of multiple immunological diseases. Inhibition of MIF activity with small molecules has proven beneficial in a number of disease models. Known small molecule MIF inhibitors typically bind in the tautomerase site of the MIF trimer, often covalently modifying the catalytic proline. Allosteric MIF inhibitors, particularly those that associate with the protein by noncovalent interactions, could reveal novel ways to block MIF activity for therapeutic benefit and serve as chemical probes to elucidate the structural basis for the diverse regulatory properties of MIF. In this study, we report the identification and functional characterization of a novel allosteric MIF inhibitor. Identified from a high throughput screening effort, this sulfonated azo compound termed p425 strongly inhibited the ability of MIF to tautomerize 4-hydroxyphenyl pyruvate. Furthermore, p425 blocked the interaction of MIF with its receptor, CD74, and interfered with the pro-inflammatory activities of the cytokine. Structural studies revealed a unique mode of binding for p425, with a single molecule of the inhibitor occupying the interface of two MIF trimers. The inhibitor binds MIF mainly on the protein surface through hydrophobic interactions that are stabilized by hydrogen bonding with four highly specific residues from three different monomers. The mode of p425 binding reveals a unique way to block the activity of the cytokine for potential therapeutic benefit in MIF-associated diseases.

  20. Modulation of hemoglobin dynamics by an allosteric effector

    PubMed Central

    Maccarini, Marco; Fouquet, Peter; Ho, Nancy T.; Ho, Chien; Makowski, Lee

    2017-01-01

    Abstract Hemoglobin (Hb) is an extensively studied paradigm of proteins that alter their function in response to allosteric effectors. Models of its action have been used as prototypes for structure‐function relationships in many proteins, and models for the molecular basis of its function have been deeply studied and extensively argued. Recent reports suggest that dynamics may play an important role in its function. Relatively little is known about the slow, correlated motions of hemoglobin subunits in various structural states because experimental and computational strategies for their characterization are challenging. Allosteric effectors such as inositol hexaphosphate (IHP) bind to both deoxy‐Hb and HbCO, albeit at different sites, leading to a lowered oxygen affinity. The manner in which these effectors impact oxygen binding is unclear and may involve changes in structure, dynamics or both. Here we use neutron spin echo measurements accompanied by wide‐angle X‐ray scattering to show that binding of IHP to HbCO results in an increase in the rate of coordinated motions of Hb subunits relative to one another with little if any change in large scale structure. This increase of large‐scale dynamics seems to be coupled with a decrease in the average magnitude of higher frequency modes of individual residues. These observations indicate that enhanced dynamic motions contribute to the functional changes induced by IHP and suggest that they may be responsible for the lowered oxygen affinity triggered by these effectors. PMID:27977887

  1. Regulation of DNA Strand Displacement Using Allosteric DNA Toehold.

    PubMed

    Yang, Xiaolong; Tang, Yanan; Traynor, Sarah M; Li, Feng

    2016-10-05

    Toehold-mediated DNA strand displacement is the fundamental basis for the construction and operation of diverse DNA devices, including circuits, machines, sensors, and reconfigurable structures. Controllable activation and regulation of toeholds are critical to construct devices with multistep, autonomous, and complex behaviors. A handful of unique toehold activation mechanisms, including toehold-exchange, associative toehold, and remote toehold, have been developed and are often combined to achieve desired strand displacement behaviors and functions. Here we report an allosteric DNA toehold (A-toehold) design that allows the flexible regulation of DNA strand displacement by splitting an input strand into an A-toehold and branch migration domain. Because of its simplicity, the A-toehold mechanism can be a useful addition to the current toolbox of DNA strand displacement techniques. We demonstrated that A-toehold enabled a number of interesting functions that were previously shown using more sophisticated DNA strand displacement systems, including 1) continuously tuning the rate of strand displacement, 2) dynamic control of strand displacement reactions, and 3) selective activation of multiple strand displacement reactions. Moreover, by combining A-toehold and toehold-exchange mechanisms, we have successfully constructed a non-covalent DNA catalysis network that resembles an allosteric enzyme.

  2. Multimodal mechanism of action of allosteric HIV-1 integrase inhibitors

    PubMed Central

    Jurado, Kellie Ann; Engelman, Alan

    2013-01-01

    Integrase (IN) is required for lentivirus replication and is a proven drug target for the prevention of AIDS in HIV-1 infected patients. While clinical strand transfer inhibitors disarm the IN active site, allosteric inhibition of enzyme activity through the disruption of IN-IN protein interfaces holds great therapeutic potential. A promising class of allosteric IN inhibitors (ALLINIs), 2-(quinolin-3-yl) acetic acid derivatives, engage the IN catalytic core domain dimerization interface at the binding site for the host integration co-factor LEDGF/p75. ALLINIs promote IN multimerization and, independent of LEDGF/p75 protein, block the formation of the active IN-DNA complex, as well as inhibit the IN-LEDGF/p75 interaction in vitro. Yet, rather unexpectedly, the full inhibitory effect of these compounds is exerted during the late phase of HIV-1 replication. ALLINIs impair particle core maturation as well as reverse transcription and integration during the subsequent round of virus infection. Recapitulating the pleiotropic phenotypes observed with numerous IN mutant viruses, ALLINIs provide insight into underlying aspects of IN biology that extend beyond its catalytic activity. Therefore, in addition to the potential to expand our repertoire of HIV-1 antiretrovirals, ALLINIs afford important structural probes to dissect the multifaceted nature of the IN protein throughout the course of HIV-1 replication. PMID:24274067

  3. Allosteric activation of ADAMTS13 by von Willebrand factor.

    PubMed

    Muia, Joshua; Zhu, Jian; Gupta, Garima; Haberichter, Sandra L; Friedman, Kenneth D; Feys, Hendrik B; Deforche, Louis; Vanhoorelbeke, Karen; Westfield, Lisa A; Roth, Robyn; Tolia, Niraj Harish; Heuser, John E; Sadler, J Evan

    2014-12-30

    The metalloprotease ADAMTS13 cleaves von Willebrand factor (VWF) within endovascular platelet aggregates, and ADAMTS13 deficiency causes fatal microvascular thrombosis. The proximal metalloprotease (M), disintegrin-like (D), thrombospondin-1 (T), Cys-rich (C), and spacer (S) domains of ADAMTS13 recognize a cryptic site in VWF that is exposed by tensile force. Another seven T and two complement C1r/C1s, sea urchin epidermal growth factor, and bone morphogenetic protein (CUB) domains of uncertain function are C-terminal to the MDTCS domains. We find that the distal T8-CUB2 domains markedly inhibit substrate cleavage, and binding of VWF or monoclonal antibodies to distal ADAMTS13 domains relieves this autoinhibition. Small angle X-ray scattering data indicate that distal T-CUB domains interact with proximal MDTCS domains. Thus, ADAMTS13 is regulated by substrate-induced allosteric activation, which may optimize VWF cleavage under fluid shear stress in vivo. Distal domains of other ADAMTS proteases may have similar allosteric properties.

  4. Allosteric Inhibition of Macrophage Migration Inhibitory Factor Revealed by Ibudilast

    SciTech Connect

    Cho, Y.; Crichlow, G; Vermeire, J; Leng, L; Du, X; Hodsdon, M; Bucala, R; Cappello, M; Gross, M; et al.

    2010-01-01

    AV411 (ibudilast; 3-isobutyryl-2-isopropylpyrazolo-[1,5-a]pyridine) is an antiinflammatory drug that was initially developed for the treatment of bronchial asthma but which also has been used for cerebrovascular and ocular indications. It is a nonselective inhibitor of various phosphodiesterases (PDEs) and has varied antiinflammatory activity. More recently, AV411 has been studied as a possible therapeutic for the treatment of neuropathic pain and opioid withdrawal through its actions on glial cells. As described herein, the PDE inhibitor AV411 and its PDE-inhibition-compromised analog AV1013 inhibit the catalytic and chemotactic functions of the proinflammatory protein, macrophage migration inhibitory factor (MIF). Enzymatic analysis indicates that these compounds are noncompetitive inhibitors of the p-hydroxyphenylpyruvate (HPP) tautomerase activity of MIF and an allosteric binding site of AV411 and AV1013 is detected by NMR. The allosteric inhibition mechanism is further elucidated by X-ray crystallography based on the MIF/AV1013 binary and MIF/AV1013/HPP ternary complexes. In addition, our antibody experiments directed against MIF receptors indicate that CXCR2 is the major receptor for MIF-mediated chemotaxis of peripheral blood mononuclear cells.

  5. New screening strategy and analysis for identification of allosteric modulators for glucagon-like peptide-1 receptor using GLP-1 (9-36) amide.

    PubMed

    Nakane, Atsushi; Gotoh, Yusuke; Ichihara, Junji; Nagata, Hidetaka

    2015-12-15

    The glucagon-like peptide-1 receptor (GLP-1R) is an important physiologic regulator of insulin secretion and a major therapeutic target for diabetes mellitus. GLP-1 (7-36) amide (active form of GLP-1) is truncated to GLP-1 (9-36) amide, which has been described as a weak agonist of GLP-1R and the major form of GLP-1 in the circulation. New classes of positive allosteric modulators (PAMs) for GLP-1R may offer improved therapeutic profiles. To identify these new classes, we developed novel and robust primary and secondary high-throughput screening (HTS) systems in which PAMs were identified to enhance the GLP-1R signaling induced by GLP-1 (9-36) amide. Screening enabled identification of two compounds, HIT-465 and HIT-736, which possessed new patterns of modulation of GLP-1R. We investigated the ability of these compounds to modify GLP-1R signaling enhanced GLP-1 (9-36) amide- and/or GLP-1 (7-36) amide-mediated cyclic adenosine monophosphate (cAMP) accumulation. These compounds also had unique profiles with regard to allosteric modulation of multiple downstream signaling (PathHunter β-arrestin signaling, PathHunter internalization signaling, microscopy-based internalization assay). We found allosteric modulation patterns to be obviously different among HIT-465, HIT-736, and Novo Nordisk compound 2. This work may enable the design of new classes of drug candidates by targeting modulation of GLP-1 (7-36) amide and GLP-1 (9-36) amide.

  6. Allosteric communication between DNA-binding and light-responsive domains of diatom class I aureochromes

    PubMed Central

    Banerjee, Ankan; Herman, Elena; Serif, Manuel; Maestre-Reyna, Manuel; Hepp, Sebastian; Pokorny, Richard; Kroth, Peter G.; Essen, Lars-Oliver; Kottke, Tilman

    2016-01-01

    The modular architecture of aureochrome blue light receptors, found in several algal groups including diatoms, is unique by having the LOV-type photoreceptor domain fused to the C-terminus of its putative effector, an N-terminal DNA-binding bZIP module. The structural and functional understanding of aureochromes’ light-dependent signaling mechanism is limited, despite their promise as an optogenetic tool. We show that class I aureochromes 1a and 1c from the diatom Phaeodactylum tricornutum are regulated in a light-independent circadian rhythm. These aureochromes are capable to form functional homo- and heterodimers, which recognize the ACGT core sequence within the canonical ‘aureo box’, TGACGT, in a light-independent manner. The bZIP domain holds a more folded and less flexible but extended conformation in the duplex DNA-bound state. FT-IR spectroscopy in the absence and the presence of DNA shows light-dependent helix unfolding in the LOV domain, which leads to conformational changes in the bZIP region. The solution structure of DNA bound to aureochrome points to a tilted orientation that was further validated by molecular dynamics simulations. We propose that aureochrome signaling relies on an allosteric pathway from LOV to bZIP that results in conformational changes near the bZIP-DNA interface without major effects on the binding affinity. PMID:27179025

  7. Potentiating mGluR5 function with a positive allosteric modulator enhances adaptive learning.

    PubMed

    Xu, Jian; Zhu, Yongling; Kraniotis, Stephen; He, Qionger; Marshall, John J; Nomura, Toshihiro; Stauffer, Shaun R; Lindsley, Craig W; Conn, P Jeffrey; Contractor, Anis

    2013-07-18

    Metabotropic glutamate receptor 5 (mGluR5) plays important roles in modulating neural activity and plasticity and has been associated with several neuropathological disorders. Previous work has shown that genetic ablation or pharmacological inhibition of mGluR5 disrupts fear extinction and spatial reversal learning, suggesting that mGluR5 signaling is required for different forms of adaptive learning. Here, we tested whether ADX47273, a selective positive allosteric modulator (PAM) of mGluR5, can enhance adaptive learning in mice. We found that systemic administration of the ADX47273 enhanced reversal learning in the Morris Water Maze, an adaptive task. In addition, we found that ADX47273 had no effect on single-session and multi-session extinction, but administration of ADX47273 after a single retrieval trial enhanced subsequent fear extinction learning. Together these results demonstrate a role for mGluR5 signaling in adaptive learning, and suggest that mGluR5 PAMs represent a viable strategy for treatment of maladaptive learning and for improving behavioral flexibility.

  8. Targeting the disordered C-terminus of PTP1B with an allosteric inhibitor

    PubMed Central

    Krishnan, Navasona; Koveal, Dorothy; Miller, Daniel H.; Xue, Bin; Akshinthala, Sai Dipikaa; Kragelj, Jaka; Jensen, Malene Ringkjøbing; Gauss, Carla-Maria; Page, Rebecca; Blackledge, Martin; Muthuswamy, Senthil K.; Peti, Wolfgang; Tonks, Nicholas K.

    2014-01-01

    PTP1B, a validated therapeutic target for diabetes and obesity, plays a critical positive role in HER2 signaling in breast tumorigenesis. Efforts to develop therapeutic inhibitors of PTP1B have been frustrated by the chemical properties of the active site. We defined a novel mechanism of allosteric inhibition that targets the C-terminal, non-catalytic segment of PTP1B. We present the first ensemble structure of PTP1B containing this intrinsically disordered segment, within which we identified a binding site for the small molecule inhibitor, MSI-1436. We demonstrate binding to a second site close to the catalytic domain, with cooperative effects between the two sites locking PTP1B in an inactive state. MSI-1436 antagonized HER2 signaling, inhibited tumorigenesis in xenografts and abrogated metastasis in the NDL2 mouse model of breast cancer, validating inhibition of PTP1B as a therapeutic strategy in breast cancer. This new approach to inhibition of PTP1B emphasizes the potential of disordered segments of proteins as specific binding sites for therapeutic small molecules. PMID:24845231

  9. Development of 1H-Pyrazolo[3,4-b]pyridines as Metabotropic Glutamate Receptor 5 Positive Allosteric Modulators.

    PubMed

    Hill, Matthew D; Fang, Haiquan; Brown, Jeffrey M; Molski, Thaddeus; Easton, Amy; Han, Xiaojun; Miller, Regina; Hill-Drzewi, Melissa; Gallagher, Lizbeth; Matchett, Michele; Gulianello, Michael; Balakrishnan, Anand; Bertekap, Robert L; Santone, Kenneth S; Whiterock, Valerie J; Zhuo, Xiaoliang; Bronson, Joanne J; Macor, John E; Degnan, Andrew P

    2016-12-08

    The metabotropic glutamate receptor 5 (mGluR5) is an attractive target for the treatment of schizophrenia due to its role in regulating glutamatergic signaling in association with the N-methyl-d-aspartate receptor (NMDAR). We describe the synthesis of 1H-pyrazolo[3,4-b]pyridines and their utility as mGluR5 positive allosteric modulators (PAMs) without inherent agonist activity. A facile and convergent synthetic route provided access to a structurally diverse set of analogues that contain neither the aryl-acetylene-aryl nor aryl-methyleneoxy-aryl elements, the predominant structural motifs described in the literature. Binding studies suggest that members of our new chemotype do not engage the receptor at the MPEP and CPPHA mGluR5 allosteric sites. SAR studies culminated in the first non-MPEP site PAM, 1H-pyrazolo[3,4-b]pyridine 31 (BMT-145027), to improve cognition in a preclinical rodent model of learning and memory.

  10. Structural insights into Ca2+-activated long-range allosteric channel gating of RyR1

    PubMed Central

    Wei, Risheng; Wang, Xue; Zhang, Yan; Mukherjee, Saptarshi; Zhang, Lei; Chen, Qiang; Huang, Xinrui; Jing, Shan; Liu, Congcong; Li, Shuang; Wang, Guangyu; Xu, Yaofang; Zhu, Sujie; Williams, Alan J; Sun, Fei; Yin, Chang-Cheng

    2016-01-01

    Ryanodine receptors (RyRs) are a class of giant ion channels with molecular mass over 2.2 mega-Daltons. These channels mediate calcium signaling in a variety of cells. Since more than 80% of the RyR protein is folded into the cytoplasmic assembly and the remaining residues form the transmembrane domain, it has been hypothesized that the activation and regulation of RyR channels occur through an as yet uncharacterized long-range allosteric mechanism. Here we report the characterization of a Ca2+-activated open-state RyR1 structure by cryo-electron microscopy. The structure has an overall resolution of 4.9 Å and a resolution of 4.2 Å for the core region. In comparison with the previously determined apo/closed-state structure, we observed long-range allosteric gating of the channel upon Ca2+ activation. In-depth structural analyses elucidated a novel channel-gating mechanism and a novel ion selectivity mechanism of RyR1. Our work not only provides structural insights into the molecular mechanisms of channel gating and regulation of RyRs, but also sheds light on structural basis for channel-gating and ion selectivity mechanisms for the six-transmembrane-helix cation channel family. PMID:27573175

  11. The nucleotide switch in Cdc42 modulates coupling between the GTPase-binding and allosteric equilibria of Wiskott–Aldrich syndrome protein

    PubMed Central

    Leung, Daisy W.; Rosen, Michael K.

    2005-01-01

    The GTP/GDP nucleotide switch in Ras superfamily GTPases generally involves differential affinity toward downstream effectors, with the GTP-bound state having a higher affinity for effector than the GDP-bound state. We have developed a quantitative model of allosteric regulation of the Wiskott–Aldrich syndrome protein (WASP) by the Rho GTPase Cdc42 to better understand how GTPase binding is coupled to effector activation. The model accurately predicts WASP affinity for Cdc42, activity toward Arp2/3 complex, and activation by Cdc42 as functions of a two-state allosteric equilibrium in WASP. The ratio of GTPase affinities for the inactive and active states of WASP is appreciably larger for Cdc42–GTP than for Cdc42–GDP. The greater ability to distinguish between the two states of WASP makes Cdc42–GTP a full WASP agonist, whereas Cdc42–GDP is only a partial agonist. Thus, the nucleotide switch controls not only the affinity of Cdc42 for its effector but also the efficiency of coupling between the Cdc42-binding and allosteric equilibria in WASP. This effect can ensure high fidelity and specificity in Cdc42 signaling in crowded membrane environments. PMID:15821030

  12. Allosteric Modulation of Alpha7 Nicotinic Receptors: Mechanistic Insight through Metadynamics and Essential Dynamics.

    PubMed

    Grazioso, Giovanni; Sgrignani, Jacopo; Capelli, Romina; Matera, Carlo; Dallanoce, Clelia; De Amici, Marco; Cavalli, Andrea

    2015-12-28

    Increasing attention has recently been devoted to allosteric modulators, as they can provide inherent advantages over classic receptor agonists. In the field of nicotinic receptors (nAChRs), the main advantage is that allosteric modulators can trigger pharmacological responses, limiting receptor desensitization. Most of the known allosteric ligands are "positive allosteric modulators" (PAMs), which increase both sensitivity to receptor agonists and current amplitude. Intriguingly, some allosteric modulators are also able to activate the α7 receptor (α7-nAChR) even in the absence of orthosteric agonists. These compounds have been named "ago-allosteric modulators" and GAT107 has been studied in depth because of its unique mechanism of action. We here investigate by molecular dynamics simulations, metadynamics, and essential dynamics the activation mechanism of α7-nAChR, in the presence of different nicotinic modulators. We determine the free energy profiles associated with the closed-to-open motion of the loop C, and we highlight mechanistic differences observed in the presence of different modulators. In particular, we demonstrate that GAT107 triggers conformational motions and cross-talk similar to those observed when the α7-nACh receptor is in complex with both an agonist and an allosteric modulator.

  13. Allosteric modulation in monomers and oligomers of a G protein-coupled receptor

    PubMed Central

    Shivnaraine, Rabindra V; Kelly, Brendan; Sankar, Krishana S; Redka, Dar'ya S; Han, Yi Rang; Huang, Fei; Elmslie, Gwendolynne; Pinto, Daniel; Li, Yuchong; Rocheleau, Jonathan V; Gradinaru, Claudiu C; Ellis, John; Wells, James W

    2016-01-01

    The M2 muscarinic receptor is the prototypic model of allostery in GPCRs, yet the molecular and the supramolecular determinants of such effects are unknown. Monomers and oligomers of the M2 muscarinic receptor therefore have been compared to identify those allosteric properties that are gained in oligomers. Allosteric interactions were monitored by means of a FRET-based sensor of conformation at the allosteric site and in pharmacological assays involving mutants engineered to preclude intramolecular effects. Electrostatic, steric, and conformational determinants of allostery at the atomic level were examined in molecular dynamics simulations. Allosteric effects in monomers were exclusively negative and derived primarily from intramolecular electrostatic repulsion between the allosteric and orthosteric ligands. Allosteric effects in oligomers could be positive or negative, depending upon the allosteric-orthosteric pair, and they arose from interactions within and between the constituent protomers. The complex behavior of oligomers is characteristic of muscarinic receptors in myocardial preparations. DOI: http://dx.doi.org/10.7554/eLife.11685.001 PMID:27151542

  14. Allosteric Modulation: An Alternate Approach Targeting the Cannabinoid CB1 Receptor.

    PubMed

    Nguyen, Thuy; Li, Jun-Xu; Thomas, Brian F; Wiley, Jenny L; Kenakin, Terry P; Zhang, Yanan

    2016-11-23

    The cannabinoid CB1 receptor is a G protein coupled receptor and plays an important role in many biological processes and physiological functions. A variety of CB1 receptor agonists and antagonists, including endocannabinoids, phytocannabinoids, and synthetic cannabinoids, have been discovered or developed over the past 20 years. In 2005, it was discovered that the CB1 receptor contains allosteric site(s) that can be recognized by small molecules or allosteric modulators. A number of CB1 receptor allosteric modulators, both positive and negative, have since been reported and importantly, they display pharmacological characteristics that are distinct from those of orthosteric agonists and antagonists. Given the psychoactive effects commonly associated with CB1 receptor agonists and antagonists/inverse agonists, allosteric modulation may offer an alternate approach to attain potential therapeutic benefits while avoiding inherent side effects of orthosteric ligands. This review details the complex pharmacological profiles of these allosteric modulators, their structure-activity relationships, and efforts in elucidating binding modes and mechanisms of actions of reported CB1 allosteric modulators. The ultimate development of CB1 receptor allosteric ligands could potentially lead to improved therapies for CB1-mediated neurological disorders.

  15. Allosteric Modulation of Metabotropic Glutamate Receptors: Structural Insights and Therapeutic Potential

    PubMed Central

    Gregory, Karen J.; Dong, Elizabeth N.; Meiler, Jens; Conn, P. Jeffrey

    2010-01-01

    Allosteric modulation of G protein-coupled receptors (GPCRs) represents a novel approach to the development of probes and therapeutics that is expected to enable subtype-specific regulation of central nervous system target receptors. The metabotropic glutamate receptors (mGlus) are class C GPCRs that play important neuromodulatory roles throughout the brain, as such they are attractive targets for therapeutic intervention for a number of psychiatric and neurological disorders including anxiety, depression, Fragile X Syndrome, Parkinson’s disease and schizophrenia. Over the last fifteen years, selective allosteric modulators have been identified for many members of the mGlu family. The vast majority of these allosteric modulators are thought to bind within the transmembrane-spanning domains of the receptors to enhance or inhibit functional responses. A combination of mutagenesis-based studies and pharmacological approaches are beginning to provide a better understanding of mGlu allosteric sites. Collectively, when mapped onto a homology model of the different mGlu subtypes based on the β2-adrenergic receptor, the previous mutagenesis studies suggest commonalities in the location of allosteric sites across different members of the mGlu family. In addition, there is evidence for multiple allosteric binding pockets within the transmembrane region that can interact to modulate one another. In the absence of a class C GPCR crystal structure, this approach has shown promise with respect to the interpretation of mutagenesis data and understanding structure-activity relationships of allosteric modulator pharmacophores. PMID:20637216

  16. A Transient Interaction between the Phosphate Binding Loop and Switch I Contributes to the Allosteric Network between Receptor and Nucleotide in Gαi1*

    PubMed Central

    Thaker, Tarjani M.; Sarwar, Maruf; Preininger, Anita M.; Hamm, Heidi E.; Iverson, T. M.

    2014-01-01

    Receptor-mediated activation of the Gα subunit of heterotrimeric G proteins requires allosteric communication between the receptor binding site and the guanine nucleotide binding site, which are separated by >30 Å. Structural changes in the allosteric network connecting these sites are predicted to be transient in the wild-type Gα subunit, making studies of these connections challenging. In the current work, site-directed mutants that alter the energy barriers between the activation states are used as tools to better understand the transient features of allosteric signaling in the Gα subunit. The observed differences in relative receptor affinity for intact Gαi1 subunits versus C-terminal Gαi1 peptides harboring the K345L mutation are consistent with this mutation modulating the allosteric network in the protein subunit. Measurement of nucleotide exchange rates, affinity for metarhodopsin II, and thermostability suggest that the K345L Gαi1 variant has reduced stability in both the GDP-bound and nucleotide-free states as compared with wild type but similar stability in the GTPγS-bound state. High resolution x-ray crystal structures reveal conformational changes accompanying the destabilization of the GDP-bound state. Of these, the conformation for Switch I was stabilized by an ionic interaction with the phosphate binding loop. Further site-directed mutagenesis suggests that this interaction between Switch I and the phosphate binding loop is important for receptor-mediated nucleotide exchange in the wild-type Gαi1 subunit. PMID:24596087

  17. Allosteric MEK1/2 inhibitors including cobimetanib and trametinib in the treatment of cutaneous melanomas.

    PubMed

    Roskoski, Robert

    2017-03-01

    The Ras-Raf-MEK-ERK (Map kinase) cellular pathway is a highly conserved eukaryotic signaling module that transduces extracellular signals from growth factors and cytokines into intracellular regulatory events that are involved in cell growth and proliferation or the contrary pathway of cell differentiation. Dysregulation of this pathway occurs in more than one-third of all malignancies, a process that has fostered the development of targeted Map kinase pathway inhibitors. Cutaneous melanomas, which arise from skin melanocytes, are the most aggressive form of skin cancer. Mutations that activate the Map kinase pathway occur in more than 90% of these melanomas. This has led to the development of the combination of dabrafenib and trametinib or vemurafenib and cobimetanib for the treatment of BRAF V600E mutant melanomas. Dabrafenib and vemurafenib target V600E/K BRAF mutants while trametinib and cobimetanib target MEK1/2. The latter two agents bind to MEK1/2 at a site that is adjacent to, but separate from, the ATP-binding site and are therefore classified as type III allosteric protein kinase inhibitors. These agents form a hydrogen bond with a conserved β3-lysine and they make numerous hydrophobic contacts with residues within the αC-helix, the β5 strand, and within the activation segment, regions of the protein kinase domain that exhibit greater diversity than those found within the ATP-binding site. One advantage of such allosteric inhibitors is that they do not have to compete with millimolar concentrations of cellular ATP, which most FDA-approved small molecule competitive inhibitors such as imatinib must do. Owing to the wide spread activation of this pathway in numerous neoplasms, trametinib and cobimetinib are being studied in combination with other targeted and cytotoxic drugs in a variety of clinical situations. Except for BRAF and NRAS mutations, there are no other biomarkers correlated with treatment responses following MEK1/2 inhibition and the

  18. Positive Allosteric Modulation of the Glucagon-like Peptide-1 Receptor by Diverse Electrophiles.

    PubMed

    Bueno, Ana B; Showalter, Aaron D; Wainscott, David B; Stutsman, Cynthia; Marín, Aranzazu; Ficorilli, James; Cabrera, Over; Willard, Francis S; Sloop, Kyle W

    2016-05-13

    Therapeutic intervention to activate the glucagon-like peptide-1 receptor (GLP-1R) enhances glucose-dependent insulin secretion and improves energy balance in patients with type 2 diabetes mellitus. Studies investigating mechanisms whereby peptide ligands activate GLP-1R have utilized mutagenesis, receptor chimeras, photo-affinity labeling, hydrogen-deuterium exchange, and crystallography of the ligand-binding ectodomain to establish receptor homology models. However, this has not enabled the design or discovery of drug-like non-peptide GLP-1R activators. Recently, studies investigating 4-(3-benzyloxyphenyl)-2-ethylsulfinyl-6-(trifluoromethyl)pyrimidine (BETP), a GLP-1R-positive allosteric modulator, determined that Cys-347 in the GLP-1R is required for positive allosteric modulator activity via covalent modification. To advance small molecule activation of the GLP-1R, we characterized the insulinotropic mechanism of BETP. In guanosine 5'-3-O-(thio)triphosphate binding and INS1 832-3 insulinoma cell cAMP assays, BETP enhanced GLP-1(9-36)-NH2-stimulated cAMP signaling. Using isolated pancreatic islets, BETP potentiated insulin secretion in a glucose-dependent manner that requires both the peptide ligand and GLP-1R. In studies of the covalent mechanism, PAGE fluorography showed labeling of GLP-1R in immunoprecipitation experiments from GLP-1R-expressing cells incubated with [(3)H]BETP. Furthermore, we investigated whether other reported GLP-1R activators and compounds identified from screening campaigns modulate GLP-1R by covalent modification. Similar to BETP, several molecules were found to enhance GLP-1R signaling in a Cys-347-dependent manner. These chemotypes are electrophiles that react with GSH, and LC/MS determined the cysteine adducts formed upon conjugation. Together, our results suggest covalent modification may be used to stabilize the GLP-1R in an active conformation. Moreover, the findings provide pharmacological guidance for the discovery and

  19. Positive Allosteric Modulation of the Glucagon-like Peptide-1 Receptor by Diverse Electrophiles*

    PubMed Central

    Showalter, Aaron D.; Wainscott, David B.; Stutsman, Cynthia; Marín, Aranzazu; Ficorilli, James; Cabrera, Over

    2016-01-01

    Therapeutic intervention to activate the glucagon-like peptide-1 receptor (GLP-1R) enhances glucose-dependent insulin secretion and improves energy balance in patients with type 2 diabetes mellitus. Studies investigating mechanisms whereby peptide ligands activate GLP-1R have utilized mutagenesis, receptor chimeras, photo-affinity labeling, hydrogen-deuterium exchange, and crystallography of the ligand-binding ectodomain to establish receptor homology models. However, this has not enabled the design or discovery of drug-like non-peptide GLP-1R activators. Recently, studies investigating 4-(3-benzyloxyphenyl)-2-ethylsulfinyl-6-(trifluoromethyl)pyrimidine (BETP), a GLP-1R-positive allosteric modulator, determined that Cys-347 in the GLP-1R is required for positive allosteric modulator activity via covalent modification. To advance small molecule activation of the GLP-1R, we characterized the insulinotropic mechanism of BETP. In guanosine 5′-3-O-(thio)triphosphate binding and INS1 832-3 insulinoma cell cAMP assays, BETP enhanced GLP-1(9–36)-NH2-stimulated cAMP signaling. Using isolated pancreatic islets, BETP potentiated insulin secretion in a glucose-dependent manner that requires both the peptide ligand and GLP-1R. In studies of the covalent mechanism, PAGE fluorography showed labeling of GLP-1R in immunoprecipitation experiments from GLP-1R-expressing cells incubated with [3H]BETP. Furthermore, we investigated whether other reported GLP-1R activators and compounds identified from screening campaigns modulate GLP-1R by covalent modification. Similar to BETP, several molecules were found to enhance GLP-1R signaling in a Cys-347-dependent manner. These chemotypes are electrophiles that react with GSH, and LC/MS determined the cysteine adducts formed upon conjugation. Together, our results suggest covalent modification may be used to stabilize the GLP-1R in an active conformation. Moreover, the findings provide pharmacological guidance for the discovery and

  20. In Vivo Investigation of Escitalopram’s Allosteric Site on the Serotonin Transporter

    PubMed Central

    Murray, Karen E.; Ressler, Kerry J.; Owens, Michael J.

    2015-01-01

    Escitalopram is a commonly prescribed antidepressant of the selective serotonin reuptake inhibitor class. Clinical evidence and mapping of the serotonin transporter (SERT) identified that escitalopram, in addition to its binding to a primary uptake-blocking site, is capable of binding to the SERT via an allosteric site that is hypothesized to alter escitalopram’s kinetics at the SERT. The studies reported here examined the in vivo role of the SERT allosteric site in escitalopram action. A knockin mouse model that possesses an allosteric-null SERT was developed. Autoradiographic studies indicated that the knockin protein was expressed at a lower density than endogenous mouse SERT (approximately 10–30% of endogenous mouse SERT), but the knockin mice are a viable tool to study the allosteric site. Microdialysis studies in the ventral hippocampus found no measurable decrease in extracellular serotonin response after local escitalopram challenge in mice without the allosteric site compared to mice with the site (p = 0.297). In marble burying assays there was a modest effect of the absence of the allosteric site, with a larger systemic dose of escitalopram (10-fold) necessary for the same effect as in mice with intact SERT (p = 0.023). However, there was no effect of the allosteric site in the tail suspension test. Together these data suggest that there may be a regional specificity in the role of the allosteric site. The lack of a robust effect overall suggests that the role of the allosteric site for escitalopram on the SERT may not produce meaningful in vivo effects. PMID:26621784

  1. Understanding the Functional Plasticity in Neural Networks of the Basal Ganglia in Cocaine Use Disorder: A Role for Allosteric Receptor-Receptor Interactions in A2A-D2 Heteroreceptor Complexes

    PubMed Central

    Borroto-Escuela, Dasiel O.; Wydra, Karolina; Pintsuk, Julia; Narvaez, Manuel; Corrales, Fidel; Zaniewska, Magdalena; Agnati, Luigi F.; Franco, Rafael; Tanganelli, Sergio; Filip, Malgorzata

    2016-01-01

    Our hypothesis is that allosteric receptor-receptor interactions in homo- and heteroreceptor complexes may form the molecular basis of learning and memory. This principle is illustrated by showing how cocaine abuse can alter the adenosine A2AR-dopamine D2R heterocomplexes and their receptor-receptor interactions and hereby induce neural plasticity in the basal ganglia. Studies with A2AR ligands using cocaine self-administration procedures indicate that antagonistic allosteric A2AR-D2R heterocomplexes of the ventral striatopallidal GABA antireward pathway play a significant role in reducing cocaine induced reward, motivation, and cocaine seeking. Anticocaine actions of A2AR agonists can also be produced at A2AR homocomplexes in these antireward neurons, actions in which are independent of D2R signaling. At the A2AR-D2R heterocomplex, they are dependent on the strength of the antagonistic allosteric A2AR-D2R interaction and the number of A2AR-D2R and A2AR-D2R-sigma1R heterocomplexes present in the ventral striatopallidal GABA neurons. It involves a differential cocaine-induced increase in sigma1Rs in the ventral versus the dorsal striatum. In contrast, the allosteric brake on the D2R protomer signaling in the A2AR-D2R heterocomplex of the dorsal striatopallidal GABA neurons is lost upon cocaine self-administration. This is potentially due to differences in composition and allosteric plasticity of these complexes versus those in the ventral striatopallidal neurons. PMID:27872762

  2. Allosteric properties of PH domains in Arf regulatory proteins.

    PubMed

    Roy, Neeladri Sekhar; Yohe, Marielle E; Randazzo, Paul A; Gruschus, James M

    2016-01-01

    Pleckstrin Homology (PH) domains bind phospholipids and proteins. They are critical regulatory elements of a number enzymes including guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) for Ras-superfamily guanine nucleotide binding proteins such as ADP-ribosylation factors (Arfs). Recent studies have indicated that many PH domains may bind more than one ligand cooperatively. Here we discuss the molecular basis of PH domain-dependent allosteric behavior of 2 ADP-ribosylation factor exchange factors, Grp1 and Brag2, cooperative binding of ligands to the PH domains of Grp1 and the Arf GTPase-activating protein, ASAP1, and the consequences for activity of the associated catalytic domains.

  3. Enhancing NMDA Receptor Function: Recent Progress on Allosteric Modulators

    PubMed Central

    2017-01-01

    The N-methyl-D-aspartate receptors (NMDARs) are subtype glutamate receptors that play important roles in excitatory neurotransmission and synaptic plasticity. Their hypo- or hyperactivation are proposed to contribute to the genesis or progression of various brain diseases, including stroke, schizophrenia, depression, and Alzheimer's disease. Past efforts in targeting NMDARs for therapeutic intervention have largely been on inhibitors of NMDARs. In light of the discovery of NMDAR hypofunction in psychiatric disorders and perhaps Alzheimer's disease, efforts in boosting NMDAR activity/functions have surged in recent years. In this review, we will focus on enhancing NMDAR functions, especially on the recent progress in the generation of subunit-selective, allosteric positive modulators (PAMs) of NMDARs. We shall also discuss the usefulness of these newly developed NMDAR-PAMs. PMID:28163934

  4. Engineering an allosteric transcription factor to respond to new ligands.

    PubMed

    Taylor, Noah D; Garruss, Alexander S; Moretti, Rocco; Chan, Sum; Arbing, Mark A; Cascio, Duilio; Rogers, Jameson K; Isaacs, Farren J; Kosuri, Sriram; Baker, David; Fields, Stanley; Church, George M; Raman, Srivatsan

    2016-02-01

    Genetic regulatory proteins inducible by small molecules are useful synthetic biology tools as sensors and switches. Bacterial allosteric transcription factors (aTFs) are a major class of regulatory proteins, but few aTFs have been redesigned to respond to new effectors beyond natural aTF-inducer pairs. Altering inducer specificity in these proteins is difficult because substitutions that affect inducer binding may also disrupt allostery. We engineered an aTF, the Escherichia coli lac repressor, LacI, to respond to one of four new inducer molecules: fucose, gentiobiose, lactitol and sucralose. Using computational protein design, single-residue saturation mutagenesis or random mutagenesis, along with multiplex assembly, we identified new variants comparable in specificity and induction to wild-type LacI with its inducer, isopropyl β-D-1-thiogalactopyranoside (IPTG). The ability to create designer aTFs will enable applications including dynamic control of cell metabolism, cell biology and synthetic gene circuits.

  5. Structural Basis for Allosteric Regulation of GPCRs by Sodium Ions

    SciTech Connect

    Liu, Wei; Chun, Eugene; Thompson, Aaron A.; Chubukov, Pavel; Xu, Fei; Katritch, Vsevolod; Han, Gye Won; Roth, Christopher B.; Heitman, Laura H.; IJzerman, Adriaan P.; Cherezov, Vadim; Stevens, Raymond C.

    2012-08-31

    Pharmacological responses of G protein-coupled receptors (GPCRs) can be fine-tuned by allosteric modulators. Structural studies of such effects have been limited due to the medium resolution of GPCR structures. We reengineered the human A{sub 2A} adenosine receptor by replacing its third intracellular loop with apocytochrome b{sub 562}RIL and solved the structure at 1.8 angstrom resolution. The high-resolution structure allowed us to identify 57 ordered water molecules inside the receptor comprising three major clusters. The central cluster harbors a putative sodium ion bound to the highly conserved aspartate residue Asp{sup 2.50}. Additionally, two cholesterols stabilize the conformation of helix VI, and one of 23 ordered lipids intercalates inside the ligand-binding pocket. These high-resolution details shed light on the potential role of structured water molecules, sodium ions, and lipids/cholesterol in GPCR stabilization and function.

  6. Taurine allosterically modulates flunitrazepam binding to synaptic membranes.

    PubMed

    Quinn, M R; Miller, C L

    1992-09-01

    Taurine is hypothesized to exert its inhibitory neuromodulatory effects, in part, by interaction with the GABAA receptor. Although taurine displaces GABA agonist binding to synaptic membranes, its allosteric effects on the benzodiazepine recognition site of the GABAA receptor complex is unsettled. We determined the effects of taurine on [3H]flunitrazepam (Flu) binding to well-washed, frozen-thawed synaptic membranes prepared from rat cortex. Comparative binding studies were conducted at 37 degrees C and on ice (0-4 degrees C). At 37 degrees C taurine increased Flu binding in a concentration dependent way by interaction with a bicuculline sensitive site, similar to GABA. Taurine increased Flu binding by causing a decrease in KD. The maximal effectiveness of taurine on Flu binding could not be increased further by addition of GABA. In contrast, the maximal stimulation of Flu binding by GABA was decreased by addition of taurine to the level attained by taurine alone. These mixed agonist/antagonist effects of taurine are pharmacologically specific and qualify taurine as a partial GABA agonist in this type of allosteric interaction. However, taurine causes opposite effects on Flu binding when measured at 0-4 degrees C: taurine interacts with a bicuculline insensitive site to inhibit Flu binding by increasing the KD. Taurine inhibition of Flu binding is not overcome by increasing concentrations of GABA. Although the mechanism of taurine inhibition of Flu binding at 0-4 degrees C is unclear, it may be an indirect effect of taurine interaction with membrane phospholipids.(ABSTRACT TRUNCATED AT 250 WORDS)

  7. Allosteric control in a metalloprotein dramatically alters function

    PubMed Central

    Baxter, Elizabeth Leigh; Zuris, John A.; Wang, Charles; Vo, Phu Luong T.; Axelrod, Herbert L.; Cohen, Aina E.; Paddock, Mark L.; Nechushtai, Rachel; Onuchic, Jose N.; Jennings, Patricia A.

    2013-01-01

    Metalloproteins (MPs) comprise one-third of all known protein structures. This diverse set of proteins contain a plethora of unique inorganic moieties capable of performing chemistry that would otherwise be impossible using only the amino acids found in nature. Most of the well-studied MPs are generally viewed as being very rigid in structure, and it is widely thought that the properties of the metal centers are primarily determined by the small fraction of amino acids that make up the local environment. Here we examine both theoretically and experimentally whether distal regions can influence the metal center in the diabetes drug target mitoNEET. We demonstrate that a loop (L2) 20 Å away from the metal center exerts allosteric control over the cluster binding domain and regulates multiple properties of the metal center. Mutagenesis of L2 results in significant shifts in the redox potential of the [2Fe-2S] cluster and orders of magnitude effects on the rate of [2Fe-2S] cluster transfer to an apo-acceptor protein. These surprising effects occur in the absence of any structural changes. An examination of the native basin dynamics of the protein using all-atom simulations shows that twisting in L2 controls scissoring in the cluster binding domain and results in perturbations to one of the cluster-coordinating histidines. These allosteric effects are in agreement with previous folding simulations that predicted L2 could communicate with residues surrounding the metal center. Our findings suggest that long-range dynamical changes in the protein backbone can have a significant effect on the functional properties of MPs. PMID:23271805

  8. A Random Forest Model for Predicting Allosteric and Functional Sites on Proteins.

    PubMed

    Chen, Ava S-Y; Westwood, Nicholas J; Brear, Paul; Rogers, Graeme W; Mavridis, Lazaros; Mitchell, John B O

    2016-04-01

    We created a computational method to identify allosteric sites using a machine learning method trained and tested on protein structures containing bound ligand molecules. The Random Forest machine learning approach was adopted to build our three-way predictive model. Based on descriptors collated for each ligand and binding site, the classification model allows us to assign protein cavities as allosteric, regular or orthosteric, and hence to identify allosteric sites. 43 structural descriptors per complex were derived and were used to characterize individual protein-ligand binding sites belonging to the three classes, allosteric, regular and orthosteric. We carried out a separate validation on a further unseen set of protein structures containing the ligand 2-(N-cyclohexylamino) ethane sulfonic acid (CHES).

  9. Allosteric Optical Control of a Class B G‐Protein‐Coupled Receptor

    PubMed Central

    Broichhagen, Johannes; Johnston, Natalie R.; von Ohlen, Yorrick; Meyer‐Berg, Helena; Jones, Ben J.; Bloom, Stephen R.; Rutter, Guy A.

    2016-01-01

    Abstract Allosteric regulation promises to open up new therapeutic avenues by increasing drug specificity at G‐protein‐coupled receptors (GPCRs). However, drug discovery efforts are at present hampered by an inability to precisely control the allosteric site. Herein, we describe the design, synthesis, and testing of PhotoETP, a light‐activated positive allosteric modulator of the glucagon‐like peptide‐1 receptor (GLP‐1R), a class B GPCR involved in the maintenance of glucose homeostasis in humans. PhotoETP potentiates Ca2+, cAMP, and insulin responses to glucagon‐like peptide‐1 and its metabolites following illumination of cells with blue light. PhotoETP thus provides a blueprint for the production of small‐molecule class B GPCR allosteric photoswitches, and may represent a useful tool for understanding positive cooperativity at the GLP‐1R. PMID:27059784

  10. Discovery and Characterization of Biased Allosteric Agonists of the Chemokine Receptor CXCR3.

    PubMed

    Milanos, Lampros; Brox, Regine; Frank, Theresa; Poklukar, Gašper; Palmisano, Ralf; Waibel, Reiner; Einsiedel, Jürgen; Dürr, Maximilian; Ivanović-Burmazović, Ivana; Larsen, Olav; Hjortø, Gertrud Malene; Rosenkilde, Mette Marie; Tschammer, Nuska

    2016-03-10

    In this work we report a design, synthesis, and detailed functional characterization of unique strongly biased allosteric agonists of CXCR3 that contain tetrahydroisoquinoline carboxamide cores. Compound 11 (FAUC1036) is the first strongly biased allosteric agonist of CXCR3 that selectively induces weak chemotaxis and leads to receptor internalization and the β-arrestin 2 recruitment with potency comparable to that of the chemokine CXCL11 without any activation of G proteins. A subtle structural change (addition of a methoxy group, 14 (FAUC1104)) led to a contrasting biased allosteric partial agonist that activated solely G proteins, induced chemotaxis, but failed to induce receptor internalization or β-arrestin 2 recruitment. Concomitant structure-activity relationship studies indicated very steep structure-activity relationships, which steer the ligand bias between the β-arrestin 2 and G protein pathway. Overall, the information presented provides a powerful platform for further development and rational design of strongly biased allosteric agonists of CXCR3.

  11. Strain analysis of protein structures and low dimensionality of mechanical allosteric couplings.

    PubMed

    Mitchell, Michael R; Tlusty, Tsvi; Leibler, Stanislas

    2016-10-04

    In many proteins, especially allosteric proteins that communicate regulatory states from allosteric to active sites, structural deformations are functionally important. To understand these deformations, dynamical experiments are ideal but challenging. Using static structural information, although more limited than dynamical analysis, is much more accessible. Underused for protein analysis, strain is the natural quantity for studying local deformations. We calculate strain tensor fields for proteins deformed by ligands or thermal fluctuations using crystal and NMR structure ensembles. Strains-primarily shears-show deformations around binding sites. These deformations can be induced solely by ligand binding at distant allosteric sites. Shears reveal quasi-2D paths of mechanical coupling between allosteric and active sites that may constitute a widespread mechanism of allostery. We argue that strain-particularly shear-is the most appropriate quantity for analysis of local protein deformations. This analysis can reveal mechanical and biological properties of many proteins.

  12. Crystal structure of Sulfolobus acidocaldarius aspartate carbamoyltransferase in complex with its allosteric activator CTP.

    PubMed

    De Vos, Dirk; Xu, Ying; Aerts, Tony; Van Petegem, Filip; Van Beeumen, Jozef J

    2008-07-18

    Aspartate carbamoyltransferase (ATCase) is a paradigm for allosteric regulation of enzyme activity. B-class ATCases display very similar homotropic allosteric behaviour, but differ extensively in their heterotropic patterns. The ATCase from the thermoacidophilic archaeon Sulfolobus acidocaldarius, for example, is strongly activated by its metabolic pathway's end product CTP, in contrast with Escherichia coli ATCase which is inhibited by CTP. To investigate the structural basis of this property, we have solved the crystal structure of the S. acidocaldarius enzyme in complex with CTP. Structure comparison reveals that effector binding does not induce similar large-scale conformational changes as observed for the E. coli ATCase. However, shifts in sedimentation coefficients upon binding of the bi-substrate analogue PALA show the existence of structurally distinct allosteric states. This suggests that the so-called "Nucleotide-Perturbation model" for explaining heterotropic allosteric behaviour, which is based on global conformational strain, is not a general mechanism of B-class ATCases.

  13. Structural Features of Ion Transport and Allosteric Regulation in Sodium-Calcium Exchanger (NCX) Proteins

    PubMed Central

    Giladi, Moshe; Tal, Inbal; Khananshvili, Daniel

    2016-01-01

    segments of CBD2, but not of CBD1. The extent and strength of Ca2+-dependent rigidification at CBD2 is splice-variant dependent, showing clear correlations with phenotypes of matching NCX variants. Therefore, diverse NCX variants share a common mechanism for the initial decoding of the regulatory signal upon Ca2+ binding at the interface of CBDs, whereas the allosteric message is shaped by CBD2, the dynamic features of which are dictated by the splicing segment. PMID:26903880

  14. Allosteric Regulation and Spatial Distribution of Kainate Receptors Bound to Ancillary Proteins

    PubMed Central

    Bowie, Derek; Garcia, Elizabeth P; Marshall, John; Traynelis, Stephen F; Lange, G David

    2003-01-01

    A diverse range of accessory proteins regulates the behaviour of most ligand- and voltage-gated ion channels. For glutamate receptor 6 (GluR6) kainate receptors, two unrelated proteins, concanavalin-A (Con-A) and postsynaptic density protein 95 (PSD-95), bind to extra- and intracellular domains, respectively, but are reported to exert similar effects on GluR6 desensitization behaviour. We have tested the hypothesis that distinct allosteric binding sites control GluR6 receptors via a common transduction pathway. Rapid agonist application to excised patches revealed that neither Con-A nor PSD-95 affect the onset of desensitization. The rate of desensitization elicited by 10 mm l-glutamate was similar in control (τfast= 5.5 ± 0.4 ms), Con-A-treated patches (τfast= 6.1 ± 0.5 ms) and patches containing PSD-95 and GluR6 receptors (τfast= 4.7 ± 0.6 ms). Likewise, the time course of recovery from GluR6 desensitization was similar in both control and Con-A conditions, whereas PSD-95 accelerated recovery almost twofold. Peak and steady-state (SS) dose-response relationships to glutamate were unchanged by lectin treatment (e.g. control, EC50(SS)= 31 ± 28 μmvs Con-A, EC50(SS)= 45 ± 9 μm, n= 6), suggesting that Con-A does not convert non-conducting channels with high agonist affinity into an open conformation. Instead, we demonstrate that the effects of Con-A on macroscopic responses reflect a shift in the relative contribution of different open states of the channel. In contrast, the effect of PSD-95 on recovery behaviour suggests that the association between kainate receptors and cytoskeletal proteins regulates signalling at glutamatergic synapses. Our results show that Con-A and PSD-95 regulate kainate receptors via distinct allosteric mechanisms targeting selective molecular steps in the transduction pathway. PMID:12562952

  15. Conversion of the lac repressor into an allosterically regulated transcriptional activator for mammalian cells.

    PubMed Central

    Labow, M A; Baim, S B; Shenk, T; Levine, A J

    1990-01-01

    A novel mammalian regulatory system was created by using the Escherichia coli lac repressor. The lac repressor was converted into a mammalian transcriptional activator by modifying the lac repressor coding region to include a nuclear localization signal from the simian virus 40 (SV40) large tumor antigen and the transcription activation domain from the herpes simplex virus type 1 virion protein 16. The lac activator protein (LAP) fusions were potent activators of several promoters containing lac operator sequences positioned either upstream or downstream of the transcription unit. A single lac operator allowed for transactivation, whereas multiple operators acted synergistically when separated by a small distance. Promoters containing 14 or 21 operator sequences were induced at least 1,000-fold in response to LAP, reaching levels of activity 20 to 30 times greater than that of the SV40 early promoter in HeLa cells. Activation was strongly inhibited by isopropyl-beta-D-thiogalactoside (IPTG), indicating that LAP retained the functions needed for allosteric regulation. LAP was bifunctional, also acting as a repressor of expression of an SV40 promoter containing an operator immediately downstream of the TATA box. Finally, genetic selection schemes were developed such that LAP-expressing cell lines can be generated at high frequency from either established or primary cells in culture. Images PMID:2162473

  16. Bacterial Rotary Export ATPases Are Allosterically Regulated by the Nucleotide Second Messenger Cyclic-di-GMP*

    PubMed Central

    Trampari, Eleftheria; Stevenson, Clare E. M.; Little, Richard H.; Wilhelm, Thomas; Lawson, David M.; Malone, Jacob G.

    2015-01-01

    The widespread second messenger molecule cyclic di-GMP (cdG) regulates the transition from motile and virulent lifestyles to sessile, biofilm-forming ones in a wide range of bacteria. Many pathogenic and commensal bacterial-host interactions are known to be controlled by cdG signaling. Although the biochemistry of cyclic dinucleotide metabolism is well understood, much remains to be discovered about the downstream signaling pathways that induce bacterial responses upon cdG binding. As part of our ongoing research into the role of cdG signaling in plant-associated Pseudomonas species, we carried out an affinity capture screen for cdG binding proteins in the model organism Pseudomonas fluorescens SBW25. The flagella export AAA+ ATPase FliI was identified as a result of this screen and subsequently shown to bind specifically to the cdG molecule, with a KD in the low micromolar range. The interaction between FliI and cdG appears to be very widespread. In addition to FliI homologs from diverse bacterial species, high affinity binding was also observed for the type III secretion system homolog HrcN and the type VI ATPase ClpB2. The addition of cdG was shown to inhibit FliI and HrcN ATPase activity in vitro. Finally, a combination of site-specific mutagenesis, mass spectrometry, and in silico analysis was used to predict that cdG binds to FliI in a pocket of highly conserved residues at the interface between two FliI subunits. Our results suggest a novel, fundamental role for cdG in controlling the function of multiple important bacterial export pathways, through direct allosteric control of export ATPase proteins. PMID:26265469

  17. Studying the binding interactions of allosteric agonists and antagonists of the CXCR4 receptor.

    PubMed

    Planesas, Jesús M; Pérez-Nueno, Violeta I; Borrell, José I; Teixidó, Jordi

    2015-07-01

    Several examples of allosteric modulators of GPCRs have been reported recently in the literature, but understanding their molecular mechanism presents a new challenge for medicinal chemistry. For the specific case of the cellular receptor CXCR4, it is known that pepducins (lipidated fragments of intracellular GPCR loops) such as ATI-2341 modulate CXCR4 activity agonistically via an allosteric mechanism. Moreover, there are also examples of small organic molecules such as AMD11070 and GSK812397 which may also act as allosteric antagonists. However, incomplete knowledge of the ligand-binding sites has hampered a detailed molecular understanding of how these inhibitors work. Here, we attempt to answer this question by analysing the binding interactions between the CXCR4 receptor and the above-mentioned allosteric modulators. We propose two different allosteric binding sites, one located in the intracellular loops 1, 2 and 3 (ICL1, ICL2 and ICL3) which binds the pepducin agonist ATI-2341, and the other at a subsite of the main extracellular orthosteric binding pocket between extracellular loops 1 and 2 and the N-terminus, which binds the antagonists AMD11070 and GSK812397. Allosteric interactions between the CXCR4 and ATI-2341 were predicted by combining different modeling approaches. First, a rotational blind docking search was applied and the best poses were subsequently refined using flexible docking methods and molecular dynamic simulations. For the AMD11070 and GSK812397 antagonists, the entire CXCR4 protein surface was explored by blind docking in order to define the binding region. A second docking analysis by subsites was then performed to refine the allosteric interactions. Finally, we identified the binding residues that appear to be essential for CXCR4 allosteric modulators.

  18. Endogenous vs Exogenous Allosteric Modulators in GPCRs: A dispute for shuttling CB1 among different membrane microenvironments

    NASA Astrophysics Data System (ADS)

    Stornaiuolo, Mariano; Bruno, Agostino; Botta, Lorenzo; Regina, Giuseppe La; Cosconati, Sandro; Silvestri, Romano; Marinelli, Luciana; Novellino, Ettore

    2015-10-01

    A Cannabinoid Receptor 1 (CB1) binding site for the selective allosteric modulator ORG27569 is here identified through an integrate approach of consensus pocket prediction, mutagenesis studies and Mass Spectrometry. This unprecedented ORG27569 pocket presents the structural features of a Cholesterol Consensus Motif, a cholesterol interacting region already found in other GPCRs. ORG27569 and cholesterol affects oppositely CB1 affinity for orthosteric ligands. Moreover, the rise in cholesterol intracellular level results in CB1 trafficking to the axonal region of neuronal cells, while, on the contrary, ORG27568 binding induces CB1 enrichment at the soma. This control of receptor migration among functionally different membrane regions of the cell further contributes to downstream signalling and adds a previously unknown mechanism underpinning CB1 modulation by ORG27569 , that goes beyond a mere control of receptor affinity for orthosteric ligands.

  19. Endogenous vs Exogenous Allosteric Modulators in GPCRs: A dispute for shuttling CB1 among different membrane microenvironments

    PubMed Central

    Stornaiuolo, Mariano; Bruno, Agostino; Botta, Lorenzo; Regina, Giuseppe La; Cosconati, Sandro; Silvestri, Romano; Marinelli, Luciana; Novellino, Ettore

    2015-01-01

    A Cannabinoid Receptor 1 (CB1) binding site for the selective allosteric modulator ORG27569 is here identified through an integrate approach of consensus pocket prediction, mutagenesis studies and Mass Spectrometry. This unprecedented ORG27569 pocket presents the structural features of a Cholesterol Consensus Motif, a cholesterol interacting region already found in other GPCRs. ORG27569 and cholesterol affects oppositely CB1 affinity for orthosteric ligands. Moreover, the rise in cholesterol intracellular level results in CB1 trafficking to the axonal region of neuronal cells, while, on the contrary, ORG27568 binding induces CB1 enrichment at the soma. This control of receptor migration among functionally different membrane regions of the cell further contributes to downstream signalling and adds a previously unknown mechanism underpinning CB1 modulation by ORG27569 , that goes beyond a mere control of receptor affinity for orthosteric ligands. PMID:26482099

  20. Power transmission line monitoring system

    SciTech Connect

    Seppa, T.O.

    1993-08-17

    A method for monitoring the sag of an overhead power transmission line comprising the steps of: measuring the tension of the power line; producing an electrical signal representative of the tension measurement; processing said electrical signal in accordance with a predetermined tension-sag relationship to produce a second signal which is a function of sag of the power line; transmitting said second electrical signal to a distant location in a predetermined transmission mode; receiving the second signal at the distant location whereby current in the power line is adjusted in accordance with the received second signal.

  1. An allosteric conduit facilitates dynamic multisite substrate recognition by the SCFCdc4 ubiquitin ligase

    NASA Astrophysics Data System (ADS)

    Csizmok, Veronika; Orlicky, Stephen; Cheng, Jing; Song, Jianhui; Bah, Alaji; Delgoshaie, Neda; Lin, Hong; Mittag, Tanja; Sicheri, Frank; Chan, Hue Sun; Tyers, Mike; Forman-Kay, Julie D.

    2017-01-01

    The ubiquitin ligase SCFCdc4 mediates phosphorylation-dependent elimination of numerous substrates by binding one or more Cdc4 phosphodegrons (CPDs). Methyl-based NMR analysis of the Cdc4 WD40 domain demonstrates that Cyclin E, Sic1 and Ash1 degrons have variable effects on the primary Cdc4WD40 binding pocket. Unexpectedly, a Sic1-derived multi-CPD substrate (pSic1) perturbs methyls around a previously documented allosteric binding site for the chemical inhibitor SCF-I2. NMR cross-saturation experiments confirm direct contact between pSic1 and the allosteric pocket. Phosphopeptide affinity measurements reveal negative allosteric communication between the primary CPD and allosteric pockets. Mathematical modelling indicates that the allosteric pocket may enhance ultrasensitivity by tethering pSic1 to Cdc4. These results suggest negative allosteric interaction between two distinct binding pockets on the Cdc4WD40 domain may facilitate dynamic exchange of multiple CPD sites to confer ultrasensitive dependence on substrate phosphorylation.

  2. An allosteric conduit facilitates dynamic multisite substrate recognition by the SCFCdc4 ubiquitin ligase

    PubMed Central

    Csizmok, Veronika; Orlicky, Stephen; Cheng, Jing; Song, Jianhui; Bah, Alaji; Delgoshaie, Neda; Lin, Hong; Mittag, Tanja; Sicheri, Frank; Chan, Hue Sun; Tyers, Mike; Forman-Kay, Julie D.

    2017-01-01

    The ubiquitin ligase SCFCdc4 mediates phosphorylation-dependent elimination of numerous substrates by binding one or more Cdc4 phosphodegrons (CPDs). Methyl-based NMR analysis of the Cdc4 WD40 domain demonstrates that Cyclin E, Sic1 and Ash1 degrons have variable effects on the primary Cdc4WD40 binding pocket. Unexpectedly, a Sic1-derived multi-CPD substrate (pSic1) perturbs methyls around a previously documented allosteric binding site for the chemical inhibitor SCF-I2. NMR cross-saturation experiments confirm direct contact between pSic1 and the allosteric pocket. Phosphopeptide affinity measurements reveal negative allosteric communication between the primary CPD and allosteric pockets. Mathematical modelling indicates that the allosteric pocket may enhance ultrasensitivity by tethering pSic1 to Cdc4. These results suggest negative allosteric interaction between two distinct binding pockets on the Cdc4WD40 domain may facilitate dynamic exchange of multiple CPD sites to confer ultrasensitive dependence on substrate phosphorylation. PMID:28045046

  3. Ultrahigh Enzyme Activity Assembled in Layered Double Hydroxides via Mg(2+)-Allosteric Effector.

    PubMed

    Wang, Min; Huang, Shu-Wan; Xu, Dan; Bao, Wen-Jing; Xia, Xing-Hua

    2015-06-02

    It is well-known that some metal ions could be allosteric effectors of allosteric enzymes to activate/inhibit the catalytic activities of enzymes. In nanobiocatalytic systems constructed based on the positive metal ion-induced allosteric effect, the incorporated enzymes will be activated and thus exhibit excellent catalytic performance. Herein, we present an environmentally friendly strategy to construct a novel allosteric effect-based β-galactosidase/Mg-Al layered double hydroxide (β-gal/Mg-Al-LDH) nanobiocatalytic system via the delamination-reconstruction method. The intercalated β-gal in the LDH galleries changes its conformation significantly due to the Mg(2+)-induced allosteric interactions and other weak interactions, which causes the activation of enzymatic activity. The β-gal/Mg-Al-LDH nanobiocatalytic system shows much higher catalytic activity and affinity toward its substrate and about 30 times higher catalytic reaction velocity than the free β-gal, which suggests that Mg(2+)-induced allosteric effect plays a vital role in the improvement of enzymatic performance.

  4. Allosteric Inhibitors Have Distinct Effects, but Also Common Modes of Action, in the HCV Polymerase

    PubMed Central

    Davis, Brittny C.; Brown, Jodian A.; Thorpe, Ian F.

    2015-01-01

    The RNA-dependent RNA polymerase from the Hepatitis C Virus (gene product NS5B) is a validated drug target because of its critical role in genome replication. There are at least four distinct allosteric sites on the polymerase to which several small molecule inhibitors bind. In addition, numerous crystal structures have been solved with different allosteric inhibitors bound to the polymerase. However, the molecular mechanisms by which these small molecules inhibit the enzyme have not been fully elucidated. There is evidence that allosteric inhibitors alter the intrinsic motions and distribution of conformations sampled by the enzyme. In this study we use molecular dynamics simulations to understand the structural and dynamic changes that result when inhibitors are bound at three different allosteric binding sites on the enzyme. We observe that ligand binding at each site alters the structure and dynamics of NS5B in a distinct manner. Nonetheless, our studies also highlight commonalities in the mechanisms of action of the different inhibitors. Each inhibitor alters the conformational states sampled by the enzyme, either by rigidifying the enzyme and preventing transitions between functional conformational states or by destabilizing the enzyme and preventing functionally relevant conformations from being adequately sampled. By illuminating the molecular mechanisms of allosteric inhibition, these studies delineate the intrinsic functional properties of the enzyme and pave the way for designing novel and more effective polymerase inhibitors. This information may also be important to understand how allosteric regulation occurs in related viral polymerases and other enzymes. PMID:25863069

  5. Targeting extracellular domains D4 and D7 of vascular endothelial growth factor receptor 2 reveals allosteric receptor regulatory sites.

    PubMed

    Hyde, Caroline A C; Giese, Alexandra; Stuttfeld, Edward; Abram Saliba, Johan; Villemagne, Denis; Schleier, Thomas; Binz, H Kaspar; Ballmer-Hofer, Kurt

    2012-10-01

    Vascular endothelial growth factors (VEGFs) activate three receptor tyrosine kinases, VEGFR-1, -2, and -3, which regulate angiogenic and lymphangiogenic signaling. VEGFR-2 is the most prominent receptor in angiogenic signaling by VEGF ligands. The extracellular part of VEGF receptors consists of seven immunoglobulin homology domains (Ig domains). Earlier studies showed that domains 2 and 3 (D23) mediate ligand binding, while structural analysis of dimeric ligand/receptor complexes by electron microscopy and small-angle solution scattering revealed additional homotypic contacts in membrane-proximal Ig domains D4 and D7. Here we show that D4 and D7 are indispensable for receptor signaling. To confirm the essential role of these domains in signaling, we isolated VEGFR-2-inhibitory "designed ankyrin repeat proteins" (DARPins) that interact with D23, D4, or D7. DARPins that interact with D23 inhibited ligand binding, receptor dimerization, and receptor kinase activation, while DARPins specific for D4 or D7 did not prevent ligand binding or receptor dimerization but effectively blocked receptor signaling and functional output. These data show that D4 and D7 allosterically regulate VEGFR-2 activity. We propose that these extracellular-domain-specific DARPins represent a novel generation of receptor-inhibitory drugs for in vivo applications such as targeting of VEGFRs in medical diagnostics and for treating vascular pathologies.

  6. Targeting Extracellular Domains D4 and D7 of Vascular Endothelial Growth Factor Receptor 2 Reveals Allosteric Receptor Regulatory Sites

    PubMed Central

    Hyde, Caroline A. C.; Giese, Alexandra; Stuttfeld, Edward; Abram Saliba, Johan; Villemagne, Denis; Schleier, Thomas; Binz, H. Kaspar

    2012-01-01

    Vascular endothelial growth factors (VEGFs) activate three receptor tyrosine kinases, VEGFR-1, -2, and -3, which regulate angiogenic and lymphangiogenic signaling. VEGFR-2 is the most prominent receptor in angiogenic signaling by VEGF ligands. The extracellular part of VEGF receptors consists of seven immunoglobulin homology domains (Ig domains). Earlier studies showed that domains 2 and 3 (D23) mediate ligand binding, while structural analysis of dimeric ligand/receptor complexes by electron microscopy and small-angle solution scattering revealed additional homotypic contacts in membrane-proximal Ig domains D4 and D7. Here we show that D4 and D7 are indispensable for receptor signaling. To confirm the essential role of these domains in signaling, we isolated VEGFR-2-inhibitory “designed ankyrin repeat proteins” (DARPins) that interact with D23, D4, or D7. DARPins that interact with D23 inhibited ligand binding, receptor dimerization, and receptor kinase activation, while DARPins specific for D4 or D7 did not prevent ligand binding or receptor dimerization but effectively blocked receptor signaling and functional output. These data show that D4 and D7 allosterically regulate VEGFR-2 activity. We propose that these extracellular-domain-specific DARPins represent a novel generation of receptor-inhibitory drugs for in vivo applications such as targeting of VEGFRs in medical diagnostics and for treating vascular pathologies. PMID:22801374

  7. Remote control apparatus for transmission

    SciTech Connect

    Ebina, A.

    1989-01-10

    A remote control apparatus for a transmission is described, comprising: means for sending a signal representing an operation state of a change lever; auxiliary power means, remote-controlled by the change lever, for changing a gear position of the transmission and sending a signal representing the gear position; and control means for controlling an operation of the auxiliary power means in accordance with the change lever operation state signal and gear position signal, the control means being provided with neutral position holding means comprises signal transmission delay means. This comprises means for detecting that the shift path on which the striker presently exists is different from the shift path instructed according to the change lever operating signal, then detecting that the striker has reached the first neutral position according to the neutral position signal and generating a neutral position detection signal.

  8. Generation and transmission of 3 × 3 W-Band multi-input multi-output orthogonal frequency division multiplexing-radio-over-fiber signals using micro-ring resonators.

    PubMed

    Alavi, S E; Amiri, I S; Ahmad, H; Supa'at, A S M; Fisal, N

    2014-12-01

    Using the micro-ring resonator (MRR) system, the single and multi-carriers at frequencies of f(1)=192.898, f(2)=192.990, f(3)=193.1, f(4)=193.315, and f(5)=193.537  THz with a free spectral range (FSR) of 92, 110, 215, and 222 GHz, respectively, are generated to be suitable for a radio-over-fiber (RoF) system based on multi-input multi-output (MIMO) with orthogonal frequency division multiplexing (OFDM). Demonstrated are the concepts of all-optical MIMO signal generation and its transmission over a 50 km single mode fiber (SMF) optical link and an up to 3 m wireless link. Sixty-four multi-carriers are used in the all-optical generation of three MIMO W-Band RF signals, where the single carriers (f(3)-f(5)) transport the signals over the RoF link. The bit error rate (BER) of the overall system performance is discussed; thus, the transmission of MIMO signals is feasible for up to an SMF path 50 km long and a wireless distance of 3 m.

  9. Automotive transmission linkage system

    SciTech Connect

    Yen, F.Y.; Ardayfio, D.D.

    1990-10-02

    This patent describes a system for converting a manual multi-speed transmission to operate as an automatic-manual transmission, the system being disposed within a vehicle, the transmission having a conventional gearshift lever and a clutch pedal. It comprises: a gearshift position selection panel, the panel being located apart from the gearshift lever, the panel enabling a driver to select the desired gear position of the gearshift lever by manipulating the panel without the necessity of the driver handling the gearshift lever; a controller which transforms the gearshift selection of the driver to an output signal, the signal corresponding to the gearshift position selected by the driver; and actuating means for mechanically repositioning the gearshift lever automatically in accordance with the output signal.

  10. Diversified transmission multichannel detection system

    SciTech Connect

    Tournois, P.; Engelhard, P.

    1984-07-03

    A detection system for imaging by sonar or radar signals. The system associates diversified transmissions with an interferometric base. This base provides an angular channel formation means and each signal formed in this way is processed by matched filtering in a circuit containing copy signals characterizing the space coloring obtained by the diversified transmission means. The invention is particularly applicable to side or front looking detection sonars.

  11. Salvinorin A: allosteric interactions at the mu-opioid receptor.

    PubMed

    Rothman, Richard B; Murphy, Daniel L; Xu, Heng; Godin, Jonathan A; Dersch, Christina M; Partilla, John S; Tidgewell, Kevin; Schmidt, Matthew; Prisinzano, Thomas E

    2007-02-01

    Salvinorin A [(2S,4aR,6aR,7R,9S,10aS,10bR)-9-(acetyloxy)-2-(3-furanyl)-dodecahydro-6a,10b-dimethyl-4,10-dioxo-2h-naphtho[2,1-c]pyran-7-carboxylic acid methyl ester] is a hallucinogenic kappa-opioid receptor agonist that lacks the usual basic nitrogen atom present in other known opioid ligands. Our first published studies indicated that Salvinorin A weakly inhibited mu-receptor binding, and subsequent experiments revealed that Salvinorin A partially inhibited mu-receptor binding. Therefore, we hypothesized that Salvinorin A allosterically modulates mu-receptor binding. To test this hypothesis, we used Chinese hamster ovary cells expressing the cloned human opioid receptor. Salvinorin A partially inhibited [(3)H]Tyr-D-Ala-Gly-N-Me-Phe-Gly-ol (DAMGO) (0.5, 2.0, and 8.0 nM) binding with E(MAX) values of 78.6, 72.1, and 45.7%, respectively, and EC(50) values of 955, 1124, and 4527 nM, respectively. Salvinorin A also partially inhibited [(3)H]diprenorphine (0.02, 0.1, and 0.5 nM) binding with E(MAX) values of 86.2, 64, and 33.6%, respectively, and EC(50) values of 1231, 866, and 3078 nM, respectively. Saturation binding studies with [(3)H]DAMGO showed that Salvinorin A (10 and 30 microM) decreased the mu-receptor B(max) and increased the K(d) in a dose-dependent nonlinear manner. Saturation binding studies with [(3)H]diprenorphine showed that Salvinorin A (10 and 40 microM) decreased the mu-receptor B(max) and increased the K(d) in a dose-dependent nonlinear manner. Similar findings were observed in rat brain with [(3)H]DAMGO. Kinetic experiments demonstrated that Salvinorin A altered the dissociation kinetics of both [(3)H]DAMGO and [(3)H]diprenorphine binding to mu receptors. Furthermore, Salvinorin A acted as an uncompetitive inhibitor of DAMGO-stimulated guanosine 5'-O-(3-[(35)S]thio)-triphosphate binding. Viewed collectively, these data support the hypothesis that Salvinorin A allosterically modulates the mu-opioid receptor.

  12. First demonstration of MC-EDFA-repeatered SDM transmission of 40 x 128-Gbit/s PDM-QPSK signals per core over 6,160-km 7-core MCF.

    PubMed

    Takahashi, H; Tsuritani, T; de Gabory, E L T; Ito, T; Peng, W R; Igarashi, K; Takeshima, K; Kawaguchi, Y; Morita, I; Tsuchida, Y; Mimura, Y; Maeda, K; Saito, T; Watanabe, K; Imamura, K; Sugizaki, R; Suzuki, M

    2013-01-14

    We demonstrate the first 7-core multicore erbium-doped fiber amplified (MC-EDFA) transmission of 40 x 128-Gbit/s PDM-QPSK signals over 6,160-km 7-core multicore fiber (MCF). The crosstalk (XT) from all of the other 6 cores of a MC-EDFA and a 55-km length MCF are about -46.5 dB and -45.6 dB at center core, respectively. The core-to-core rotation approach at every amplified span is used to average the XT of all cores. The averaged optical signal-to-noise ratio (OSNR) after 6,160-km transmission is 15.6 dB with 0.1 nm resolution bandwidth. The Q-factor of all 40 channels surpasses the threshold of the forward-error-correction of 6.4 dB with 1 dB margin after 6,160 km. The total net capacity is 28.8 Tbit/s per fiber and achieved capacity-distance product is 177 Pbit/s.km per fiber. We confirmed the feasibility of MC-EDFA repeatered systems for trans-oceanic transmission.

  13. The GABAB Positive Allosteric Modulator, ADX71441 Attenuates Alcohol Self-Administration and Relapse to Alcohol Seeking in Rats.

    PubMed

    Augier, Eric; Dulman, Russell S; Damadzic, Ruslan; Pilling, Andrew; Hamilton, J Paul; Heilig, Markus

    2017-03-15

    GABAergic signaling is involved in modulating the reinforcing properties of alcohol, and GABAB receptors have been proposed as a potential target for clinical treatment of alcoholism. The orthosteric GABAB receptor agonist baclofen has been shown to suppress operant self-administration of alcohol in animals and alcohol use in alcohol dependent patients, but its utility is limited by a narrow therapeutic index. We tested the effects of ADX71441, a novel GABAB receptor positive allosteric modulator on alcohol-related behaviors in rats. We first assessed the effects of ADX71441 (1, 3, 10 and 30 mg/kg, I.P.) on both non-dependent and dependent male Wistar rats trained to self-administer 20% alcohol. We then determined the effects of ADX71441 on stress-induced as well as cue-induced relapse-like behavior. Finally, we sought to identify the brain regions through which ADX71441 may act to prevent relapse-like behavior by mapping the neuronal activation induced by stress-induced reinstatement of alcohol-seeking using c-Fos immunohistochemistry. ADX71441 dose-dependently decreased alcohol self-administration of both dependent and non-dependent animals, but its potency was higher in alcohol-dependent rats. Furthermore, both cue- and stress-induced alcohol seeking were blocked by the GABAB receptor positive allosteric modulator. Finally, pretreatment with 3 mg/kg of ADX71441 before stress-induced reinstatement significantly decreased c-Fos expression in a network of brain regions implicated in stress induced relapse, comprising the nucleus accumbens shell, the dorsal raphe nucleus and the medial prefrontal cortex. Our findings support a causal role of GABAB receptors in alcohol reinforcement and relapse to alcohol seeking. These effects are observed in the absence of significant sedative side effects. Jointly, these observations indicate that GABAB receptor positive allosteric modulators merit being tested clinically for the treatment of alcoholism. Our data also point to

  14. Structure-based discovery of the first allosteric inhibitors of cyclin-dependent kinase 2.

    PubMed

    Rastelli, Giulio; Anighoro, Andrew; Chripkova, Martina; Carrassa, Laura; Broggini, Massimo

    2014-01-01

    Allosteric targeting of protein kinases via displacement of the structural αC helix with type III allosteric inhibitors is currently gaining a foothold in drug discovery. Recently, the first crystal structure of CDK2 with an open allosteric pocket adjacent to the αC helix has been described, prospecting new opportunities to design more selective inhibitors, but the structure has not yet been exploited for the structure-based design of type III allosteric inhibitors. In this work we report the results of a virtual screening campaign that resulted in the discovery of the first-in-class type III allosteric ligands of CDK2. Using a combination of docking and post-docking analyses made with our tool BEAR, 7 allosteric ligands (hit rate of 20%) with micromolar affinity for CDK2 were identified, some of them inhibiting the growth of breast cancer cell lines in the micromolar range. Competition experiments performed in the presence of the ATP-competitive inhibitor staurosporine confirmed that the 7 ligands are truly allosteric, in agreement with their design. Of these, compound 2 bound CDK2 with an EC50 value of 3 μM and inhibited the proliferation of MDA-MB231 and ZR-75-1 breast cancer cells with IC50 values of approximately 20 μM, while compound 4 had an EC50 value of 71 μM and IC50 values around 4 μM. Remarkably, the most potent compound 4 was able to selectively inhibit CDK2-mediated Retinoblastoma phosphorylation, confirming that its mechanism of action is fully compatible with a selective inhibition of CDK2 phosphorylation in cells. Finally, hit expansion through analog search of the most potent inhibitor 4 revealed an additional ligand 4g with similar in vitro potency on breast cancer cells.

  15. ASSESSMENT OF SUBUNIT-DEPENDENT DIRECT GATING AND ALLOSTERIC MODULATORY EFFECTS OF CARISOPRODOL AT GABAA RECEPTORS

    PubMed Central

    Kumar, Manoj; González, Lorie A.; Dillon, Glenn H.

    2016-01-01

    Carisoprodol is a widely prescribed muscle relaxant, abuse of which has grown considerably in recent years. It directly activates and allosterically modulates α1β2γ2 GABAARs, although the site(s) of action are unknown. To gain insight into the actions of carisoprodol, subunit-dependent effects of this drug were assessed. Whole-cell patch clamp recordings were obtained from HEK293 cells expressing α1β2, α1β3 or αxβzγ2 (where x = 1–6 and z = 1–3) GABAARs, and in receptors incorporating the δ subunit (modeling extrasynaptic receptors). The ability to directly gate and allosterically potentiate GABA-gated currents was observed for all configurations. Presence or absence of the γ2 subunit did not affect the ability of carisoprodol to directly gate or allosterically modulate the receptor. Presence of the β1 subunit conferred highest efficacy for direct activation relative to maximum GABA currents, while presence of the β2 subunit conferred highest efficacy for allosteric modulation of the GABA response. With regard to α subunits, carisoprodol was most efficacious at enhancing the actions of GABA in receptors incorporating the α1 subunit. The ability to directly gate the receptor was generally comparable regardless of the α subunit isoform, although receptors incorporating the α3 subunit showed significantly reduced direct gating efficacy and affinity. In extrasynaptic (α1β3δ and α4β3δ) receptors, carisoprodol had greater efficacy than GABA as a direct gating agonist. In addition, carisoprodol allosterically potentiated both EC20 and saturating GABA concentrations in these receptors. In assessing voltage-dependence, we found direct gating and inhibitory effects were insensitive to membrane voltage, whereas allosteric modulatory effects were affected by membrane voltage. Our findings demonstrate direct and allosteric effects of carisoprodol at synaptic and extrasynpatic GABAARs and that subunit isoform influences these effects. PMID:25896767

  16. Allosteric receptor activation by the plant peptide hormone phytosulfokine.

    PubMed

    Wang, Jizong; Li, Hongju; Han, Zhifu; Zhang, Heqiao; Wang, Tong; Lin, Guangzhong; Chang, Junbiao; Yang, Weicai; Chai, Jijie

    2015-09-10

    Phytosulfokine (PSK) is a disulfated pentapeptide that has a ubiquitous role in plant growth and development. PSK is perceived by its receptor PSKR, a leucine-rich repeat receptor kinase (LRR-RK). The mechanisms underlying the recognition of PSK, the activation of PSKR and the identity of the components downstream of the initial binding remain elusive. Here we report the crystal structures of the extracellular LRR domain of PSKR in free, PSK- and co-receptor-bound forms. The structures reveal that PSK interacts mainly with a β-strand from the island domain of PSKR, forming an anti-β-sheet. The two sulfate moieties of PSK interact directly with PSKR, sensitizing PSKR recognition of PSK. Supported by biochemical, structural and genetic evidence, PSK binding enhances PSKR heterodimerization with the somatic embryogenesis receptor-like kinases (SERKs). However, PSK is not directly involved in PSKR-SERK interaction but stabilizes PSKR island domain for recruitment of a SERK. Our data reveal the structural basis for PSKR recognition of PSK and allosteric activation of PSKR by PSK, opening up new avenues for the design of PSKR-specific small molecules.

  17. Allosteric Small-Molecule Inhibitors of the AKT Kinase

    NASA Astrophysics Data System (ADS)

    Dalafave, D. S.

    This research addresses computational design of small druglike molecules for possible anticancer applications. AKT and SGK are kinases that control important cellular functions. They are highly homologous, having similar activators and targets. Cancers with increased SGK activity may develop resistance to AKT-specific inhibitors. Our goal was to design new molecules that would bind both AKT and SGK, thus preventing the development of drug resistance. Most kinase inhibitors target the kinase ATP-binding site. However, the high similarity in this site among kinases makes it difficult to target specifically. Furthermore, mutations in this site can cause resistance to ATP-competitive kinase inhibitors. We used existing AKT inhibitors as initial templates to design molecules that could potentially bind the allosteric sites of both AKT and SGK. Molecules with no implicit toxicities and optimal drug-like properties were used for docking studies. Binding energies of the stable complexes that the designed molecules formed with AKT and SGK were calculated. Possible applications of the designed putative inhibitors against cancers with overexpressed AKT/SGK is discussed.

  18. Allosteric activation of apicomplexan calcium-dependent protein kinases

    PubMed Central

    Ingram, Jessica R.; Knockenhauer, Kevin E.; Markus, Benedikt M.; Mandelbaum, Joseph; Ramek, Alexander; Shan, Yibing; Shaw, David E.; Schwartz, Thomas U.; Ploegh, Hidde L.; Lourido, Sebastian

    2015-01-01

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7–kinase interactions. In contrast to other Ca2+-regulated kinases, the regulatory domain of TgCDPK1 plays a dual role, inhibiting or activating the kinase in response to changes in Ca2+ concentrations. We propose that the regulatory domain of TgCDPK1 acts as a molecular splint to stabilize the otherwise inactive KD. This dependence on allosteric stabilization reveals a novel susceptibility in this important class of parasite enzymes. PMID:26305940

  19. The allosteric mechanism of yeast chorismate mutase: a dynamic analysis.

    PubMed

    Kong, Yifei; Ma, Jianpeng; Karplus, Martin; Lipscomb, William N

    2006-02-10

    The effector-regulated allosteric mechanism of yeast chorismate mutase (YCM) was studied by normal mode analysis and targeted molecular dynamics. The normal mode analysis shows that the conformational change between YCM in the R state and in the T state can be represented by a relatively small number of low-frequency modes. This suggests that the transition is coded in the structure and is likely to have a low energetic barrier. Quantitative comparisons (i.e. frequencies) between the low-frequency modes of YCM with and without effectors (modeled structures) reveal that the binding of Trp increases the global flexibility, whereas Tyr decreases global flexibility. The targeted molecular dynamics simulation of substrate analog release from the YCM active site suggests that a series of residues are critical for orienting and "recruiting" the substrate. The simulation led to the switching of a series of substrate-release-coupled salt-bridge partners in the ligand-binding domain; similar changes occur in the transition between YCM R-state and T-state crystal structures. Thus, the normal mode analysis and targeted molecular dynamics results provide evidence that the effectors regulate YCM activity by influencing the global flexibility. The change in flexibility is coupled to the binding of substrate to the T state and release of the product from the R state, respectively.

  20. Allosteric Inhibition of Anti-Apoptotic MCL-1

    PubMed Central

    Lee, Susan; Wales, Thomas E.; Escudero, Silvia; Cohen, Daniel T.; Luccarelli, James; Gallagher, Catherine; Cohen, Nicole A.; Huhn, Annissa J.; Bird, Gregory H.; Engen, John R.; Walensky, Loren D.

    2016-01-01

    MCL-1 is an anti-apoptotic BCL-2 family protein that has emerged as a major pathogenic factor in human cancer. Like BCL-2, MCL-1 bears a surface groove whose function is to sequester the BH3 killer domains of pro-apoptotic BCL-2 family members, a mechanism harnessed by cancer cells to establish formidable apoptotic blockades. Whereas drugging the BH3-binding groove has been achieved for BCL-2, translating this approach to MCL-1 has been challenging. Here, we report an alternative mechanism for MCL-1 inhibition by small molecule covalent modification of C286 at a novel interaction site distant from the BH3-binding groove. Our structure-function analyses revealed that the BH3-binding capacity of MCL-1 and its suppression of BAX are impaired by molecular engagement, a phenomenon recapitulated by C286W mutagenic mimicry in vitro and in cells. Thus, we characterize an allosteric mechanism for disrupting the anti-apoptotic, BH3-binding activity of MCL-1, informing a new strategy for disarming MCL-1 in cancer. PMID:27159560

  1. Coarse-grained molecular simulations of allosteric cooperativity

    NASA Astrophysics Data System (ADS)

    Nandigrami, Prithviraj; Portman, John J.

    2016-03-01

    Interactions between a protein and a ligand are often accompanied by a redistribution of the population of thermally accessible conformations. This dynamic response of the protein's functional energy landscape enables a protein to modulate binding affinities and control binding sensitivity to ligand concentration. In this paper, we investigate the structural origins of binding affinity and allosteric cooperativity of binding two Ca2+ ions to each domain of Calmodulin (CaM) through simulations of a simple coarse-grained model. In this model, the protein's conformational transitions between open and closed conformational ensembles are simulated explicitly and ligand binding and unbinding are treated implicitly within the grand canonical ensemble. Ligand binding is cooperative because the binding sites are coupled through a shift in the dominant conformational ensemble upon binding. The classic Monod-Wyman-Changeux model of allostery with appropriate binding free energies to the open and closed ensembles accurately describes the simulated binding thermodynamics. The simulations predict that the two domains of CaM have distinct binding affinity and cooperativity. In particular, the C-terminal domain binds Ca2+ with higher affinity and greater cooperativity than the N-terminal domain. From a structural point of view, the affinity of an individual binding loop depends sensitively on the loop's structural compatibility with the ligand in the bound ensemble, as well as the conformational flexibility of the binding site in the unbound ensemble.

  2. The therapeutic promise of positive allosteric modulation of nicotinic receptors

    PubMed Central

    Uteshev, Victor V.

    2014-01-01

    In the central nervous system, deficits in cholinergic neurotransmission correlate with decreased attention and cognitive impairment, while stimulation of neuronal nicotinic acetylcholine receptors improves attention, cognitive performance and neuronal resistance to injury as well as produces robust analgesic and anti-inflammatory effects. The rational basis for the therapeutic use of orthosteric agonists and positive allosteric modulators (PAMs) of nicotinic receptors arises from the finding that functional nicotinic receptors are ubiquitously expressed in neuronal and non-neuronal tissues including brain regions highly vulnerable to traumatic and ischemic types of injury (e.g., cortex and hippocampus). Moreover, functional nicotinic receptors do not vanish in age-, disease- and trauma-related neuropathologies, but their expression and/or activation levels decline in a subunit- and brain region-specific manner. Therefore, augmenting the endogenous cholinergic tone by nicotinic agents is possible and may offset neurological impairments associated with cholinergic hypofunction. Importantly, because neuronal damage elevates extracellular levels of choline (a selective agonist of α7 nicotinic acetylcholine receptors) near the site of injury, α7-PAM-based treatments may augment pathology-activated α7-dependent auto-therapies where and when they are most needed (i.e., in the penumbra, post-injury). Thus, the nicotinic-PAM-based treatments are expected to be highly efficacious with fewer side effects as compared to a more indiscriminate action of exogenous orthosteric agonists. In this review, I will summarize the existing trends in therapeutic applications of nicotinic PAMs. PMID:24530419

  3. Allosteric activation of apicomplexan calcium-dependent protein kinases

    SciTech Connect

    Ingram, Jessica R.; Knockenhauer, Kevin E.; Markus, Benedikt M.; Mandelbaum, Joseph; Ramek, Alexander; Shan, Yibing; Shaw, David E.; Schwartz, Thomas U.; Ploegh, Hidde L.; Lourido, Sebastian

    2015-08-24

    Calcium-dependent protein kinases (CDPKs) comprise the major group of Ca2+-regulated kinases in plants and protists. It has long been assumed that CDPKs are activated, like other Ca2+-regulated kinases, by derepression of the kinase domain (KD). However, we found that removal of the autoinhibitory domain from Toxoplasma gondii CDPK1 is not sufficient for kinase activation. From a library of heavy chain-only antibody fragments (VHHs), we isolated an antibody (1B7) that binds TgCDPK1 in a conformation-dependent manner and potently inhibits it. We uncovered the molecular basis for this inhibition by solving the crystal structure of the complex and simulating, through molecular dynamics, the effects of 1B7–kinase interactions. In contra