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1

Expression of Water Channel Proteins in Mesembryanthemum crystallinum  

Microsoft Academic Search

We have characterized transcripts for nine major intrinsic proteins (MIPs), some of which function as water channels (aquaporins), from the ice plant Mesembryanthemum crystallinum. To determine the cellular distribution and expression of these MIPs, oligopeptide-based antibodies were generated against MIP-A, MIP-B, MIP-C, or MIP-F, which, according to sequence and functional characteristics, are located in the plasma membrane (PM) and tonoplast,

Hans-Hubert Kirch; Rosario Vera-Estrella; Dortje Golldack; Francoise Quigley; Christine B. Michalowski; Bronwyn J. Barkla; Hans J. Bohnert

2000-01-01

2

The aquaporin family of water channel proteins in clinical medicine.  

PubMed

The aquaporins are a family of membrane channel proteins that serve as selective pores through which water crosses the plasma membranes of many human tissues and cell types. The sites where aquaporins are expressed implicate these proteins in renal water reabsorption, cerebrospinal fluid secretion and reabsorption, generation of pulmonary secretions, aqueous humor secretion and reabsorption, lacrimation, and multiple other physiologic processes. Determination of the aquaporin gene sequences and their chromosomal locations has provided insight into the structure and pathophysiologic roles of these proteins, and primary and secondary involvement of aquaporins is becoming apparent in diverse clinical disorders. Aquaporin-1 (AQP1) is expressed in multiple tissues including red blood cells, and the Colton blood group antigens represent a polymorphism on the AQP1 protein. AQP2 is restricted to renal collecting ducts and has been linked to congenital nephrogenic diabetes insipidus in humans and to lithium-induced nephrogenic diabetes insipidus and fluid retention from congestive heart failure in rat models. Congenital cataracts result from mutations in the mouse gene encoding the lens homolog Aqp0 (Mip). The present understanding of aquaporin physiology is still incomplete; identification of additional members of the aquaporin family will affect future studies of multiple disorders of water distribution throughout the body. In some tissues, the aquaporins may participate in the transepithelial movement of fluid without being rate limiting, so aquaporins may be involved in clinical disorders without being causative. As outlined in this review, our challenge is to identify disease states in which aquaporins are involved, to define the aquaporins' roles mechanistically, and to search for ways to exploit this information therapeutically. PMID:9193450

Lee, M D; King, L S; Agre, P

1997-05-01

3

Role of the Dielectric Constants of Membrane Proteins and Channel Water in Ion Permeation  

Microsoft Academic Search

Using both analytical solutions obtained from simplified systems and numerical results from more realistic cases, we investigate the role played by the dielectric constant of membrane proteins ?p and pore water ?w in permeation of ions across channels. We show that the boundary and its curvature are the crucial factors in determining how an ion's potential energy depends on the

Turgut Ba?tu?; Serdar Kuyucak

2003-01-01

4

Molecular cloning, overexpression and characterization of a novel water channel protein from Rhodobacter sphaeroides.  

PubMed

Aquaporins are highly selective water channel proteins integrated into plasma membranes of single cell organisms; plant roots and stromae; eye lenses, renal and red blood cells in vertebrates. To date, only a few microbial aquaporins have been characterized and their physiological importance is not well understood. Here we report on the cloning, expression and characterization of a novel aquaporin, RsAqpZ, from a purple photosynthetic bacterium, Rhodobacter sphaeroides ATCC 17023. The protein was expressed homologously at a high yield (?20 mg/L culture) under anaerobic photoheterotrophic growth conditions. Stopped-flow light scattering experiments demonstrated its high water permeability (0.17±0.05 cm/s) and low energy of activation for water transport (2.93±0.60 kcal/mol) in reconstituted proteoliposomes at a protein to lipid ratio (w/w) of 0.04. We developed a fluorescence correlation spectroscopy based technique and utilized a fluorescent protein fusion of RsAqpZ, to estimate the single channel water permeability of RsAqpZ as 1.24 (±0.41) x 10(-12) cm(3)/s or 4.17 (±1.38)×10(10) H2O molecules/s, which is among the highest single channel permeability reported for aquaporins. Towards application to water purification technologies, we also demonstrated functional incorporation of RsAqpZ in amphiphilic block copolymer membranes. PMID:24497982

Erbakan, Mustafa; Shen, Yue-xiao; Grzelakowski, Mariusz; Butler, Peter J; Kumar, Manish; Curtis, Wayne R

2014-01-01

5

Highly permeable polymeric membranes based on the incorporation of the functional water channel protein Aquaporin Z  

PubMed Central

The permeability and solute transport characteristics of amphiphilic triblock-polymer vesicles containing the bacterial water-channel protein Aquaporin Z (AqpZ) were investigated. The vesicles were made of a block copolymer with symmetric poly-(2-methyloxazoline)-poly-(dimethylsiloxane)-poly-(2-methyloxazoline) (PMOXA15-PDMS110-PMOXA15) repeat units. Light-scattering measurements on pure polymer vesicles subject to an outwardly directed salt gradient in a stopped-flow apparatus indicated that the polymer vesicles were highly impermeable. However, a large enhancement in water productivity (permeability per unit driving force) of up to ?800 times that of pure polymer was observed when AqpZ was incorporated. The activation energy (Ea) of water transport for the protein-polymer vesicles (3.4 kcal/mol) corresponded to that reported for water-channel-mediated water transport in lipid membranes. The solute reflection coefficients of glucose, glycerol, salt, and urea were also calculated, and indicated that these solutes are completely rejected. The productivity of AqpZ-incorporated polymer membranes was at least an order of magnitude larger than values for existing salt-rejecting polymeric membranes. The approach followed here may lead to more productive and sustainable water treatment membranes, whereas the variable levels of permeability obtained with different concentrations of AqpZ may provide a key property for drug delivery applications.

Kumar, Manish; Grzelakowski, Mariusz; Zilles, Julie; Clark, Mark; Meier, Wolfgang

2007-01-01

6

Characterization of OsPIP2;7, a water channel protein in rice.  

PubMed

Aquaporins are water channel proteins that facilitate passage of water and other small neutral molecules across biological membranes. There are usually a large number of members of this family in higher plants, which exhibit various physiological functions and are regulated in a time-specific and particular mode. We have previously shown that a rice gene, OsPIP2;7, was generally up-regulated in roots but down-regulated in shoots at the early stage of chilling stress. Here, OsPIP2;7 was cloned and proved to be an aquaporin with high activity in Xenopus oocytes. OsPIP2;7 was localized mainly in mesophyll cells of leaves. In roots it was detected in the vascular tissues, epidermis cells and exodermis cells at the elongation zone, as well as in the epidermis cells, exodermis cells and root hair at the maturation zone. Yeast cells overexpressing OsPIP2;7 showed a higher survival rate after freeze-thaw stress. Furthermore, OsPIP2;7 enhanced the transpiration rate and tolerance to low temperature when overexpressed in rice. These results indicated that OsPIP2;7 was involved in rapid water transport and maintenance of the water balance in cells, and ultimately improves the tolerance of yeast and rice to low temperature stress. PMID:18988636

Li, Guo-Wei; Zhang, Min-Hua; Cai, Wei-Ming; Sun, Wei-Ning; Su, Wei-Ai

2008-12-01

7

Water channel proteins: from their discovery in 1985 in Cluj-Napoca, Romania, to the 2003 Nobel Prize in Chemistry.  

PubMed

Water channel proteins, later called aquaporins, are transmembrane proteins that have as their main(specific) function the water transport across biological membranes. The first water channel protein (WCP), now called aquaporin 1, was identified or "seen" in situ (hence discovered) in the human red blood cell (RBC) membrane in 1985 by Benga's group (Cluj-Napoca, Romania). This was achieved by a very selective radiolabeling of RBC membrane proteins with the water transport inhibitor [203Hg]-p-chloromercuribenzene sulfonate (PCMBS), under conditions of specific inhibition. The presence and location of the WCP was discovered among the polypeptides migrating in the region of 35-60 kDa on the electrophoretogram of RBC membrane proteins. The work was first published in 1986 in Biochemistry and Eur. J. Cell Biol. and reviewed by Benga in several articles in 1988-2004. We have thus a world priority in the discovery of the first water channel in the RBC membrane, that was re-discovered by chance by the group of Agre (Baltimore, USA) in 1988, when they isolated a new protein from the RBC membrane, nick-named CHIP28 (channel-forming integral membrane protein of 28 kDa). However, in addition to the 28 kDa component, this protein had a 35-60 kDa glycosylated component, the one detected by Benga's group. Only in 1992 the Agre's group suggested that "it is likely that CHIP28 is a functional unit of membrane water channels". In 1993 CHIP28 was renamed aquaporin 1. Looking in retrospect, asking the crucial question, when was the first WCP, discovered, a fair and clear cut answer would be: the first WCP, now called aquaporin 1, was identified or "seen" (hence discovered) in situ in the human RBC membrane by Benga and coworkers in 1985. It was again "seen" when it was purified in 1988 and again identified when its water transport property was found byAgre's group in 1992. If we make a comparison with the discovery of New World of America, the first man who has "seen" a part, very small indeed, of The New Land was Columbus; later, others, including Amerigo Vespucci (from whom the name derived), have better "seen" and in the subsequent years many explorers discovered the complexity of the Americas. Consequently, the initial discovery of the first water channel by Benga's group must be properly credited; the omission of Gheorghe Benga from the 2003 Nobel Prize in Chemistry (half of which was awarded to Peter Agre "for the discovery of the water channels") was a new mistake in the award of Nobel Prizes. Benga's claim is presented on the web site of the Ad Astra Association (www.ad-astra.ro/benga). As can be seen on this site his recognition as a discoverer of the first water channel protein from the human RBC membrane is growing. Thousands of science-related professionals from hundreds of academic and research units, as well as participants in several international scientific events, have signed as supporters of Benga; his priority is also mentioned in several comments on the 2003 Nobel Prize as presented on the site. PMID:17543216

Benga, Gh

2006-01-01

8

Water channel proteins in bile formation and flow in health and disease: when immiscible becomes miscible.  

PubMed

An essential function of the liver is the formation and secretion of bile, a complex aqueous solution of organic and inorganic compounds essential as route for the elimination of body cholesterol as unesterified cholesterol or as bile acids. In bile, a considerable amount of otherwise insoluble cholesterol is solubilized by carriers including two other classes of lipids, namely phospholipid and bile acids. Formation of bile and generation of bile flow are driven by the active secretion of bile acids, lipids and electrolytes into the canalicular and bile duct lumens followed by the parallel movement of water. Thus, water has to cross rapidly into and out of the cell interior driven by osmotic forces. Bile as a fluid, results from complicated interplay of hepatocyte and cholangiocyte uptake and secretion, concentration, by involving a number of transporters of lipids, anions, cations, and water. The discovery of the aquaporin water channels, has clarified the mechanisms by which water, the major component of bile (more than 95%), moves across the hepatobiliary epithelia. This review is focusing on novel acquisitions in liver membrane lipidic and water transport and functional participation of aquaporin water channels in multiple aspects of hepatobiliary fluid balance. Involvement of aquaporins in a series of clinically relevant hepatobiliary disorders are also discussed. PMID:22487565

Portincasa, Piero; Calamita, Giuseppe

2012-01-01

9

Conformational Changes of an Ion Channel Detected Through Water-Protein Interactions Using Solid-State NMR Spectroscopy  

PubMed Central

The influenza A virus M2 protein is a pH-gated and amantadine-inhibited proton channel important for the virus life cycle. Proton conduction by M2 is known to involve water, however direct experimental evidence of M2-water interaction is scarce. Using 1H spin diffusion solid-state NMR, we have now determined the water accessibility of the M2 transmembrane domain (M2-TM) in virus-envelope-mimetic lipid membranes and its changes with environment. Site-specific water-protein magnetization transfer indicates that, in the absence of amantadine, the initial spin diffusion rate mainly depends on the radial position of the residues from the pore: pore-lining residues along the helix have similarly high water accessibilities compared to lipid-facing residues. Upon drug binding, the spin diffusion rates become much slower for Gly34 in the middle of the helix than for the N-terminal residues, indicating that amantadine is bound to the pore lumen between Gly34 and Val27. Water-protein spin diffusion buildup curves indicate that spin diffusion is the fastest in the low-pH open state, slower in the high-pH closed state, and the slowest in the high-pH amantadine-bound state. Simulations of the buildup curves using a 3D lattice model yielded quantitative values of the water-accessible surface area and its changes by pH and drug binding. These data provide direct experimental evidence of the pH-induced change of the pore size and the drug-induced dehydration of the pore. This study demonstrates the capability of 1H spin diffusion NMR for elucidating water interactions with ion channels, water pores, and proton pumps, and for probing membrane protein conformational changes that involve significant changes of water-accessible surface areas.

Luo, Wenbin; Hong, Mei

2010-01-01

10

Ferritin Protein Nanocage Ion Channels  

PubMed Central

Ferritin protein nanocages, self-assembled from four-?-helix bundle subunits, use Fe2+ and oxygen to synthesize encapsulated, ferric oxide minerals. Ferritin minerals are iron concentrates stored for cell growth. Ferritins are also antioxidants, scavenging Fenton chemistry reactants. Channels for iron entry and exit consist of helical hairpin segments surrounding the 3-fold symmetry axes of the ferritin nanocages. We now report structural differences caused by amino acid substitutions in the Fe2+ ion entry and exit channels and at the cytoplasmic pores, from high resolution (1.3–1.8 ?) protein crystal structures of the eukaryotic model ferritin, frog M. Mutations that eliminate conserved ionic or hydrophobic interactions between Arg-72 and Asp-122 and between Leu-110 and Leu-134 increase flexibility in the ion channels, cytoplasmic pores, and/or the N-terminal extensions of the helix bundles. Decreased ion binding in the channels and changes in ordered water are also observed. Protein structural changes coincide with increased Fe2+ exit from dissolved, ferric minerals inside ferritin protein cages; Fe2+ exit from ferritin cages depends on a complex, surface-limited process to reduce and dissolve the ferric mineral. High concentrations of bovine serum albumin or lysozyme (protein crowders) to mimic the cytoplasm restored Fe2+ exit in the variants to wild type. The data suggest that fluctuations in pore structure control gating. The newly identified role of the ferritin subunit N-terminal extensions in gating Fe2+ exit from the cytoplasmic pores strengthens the structural and functional analogies between ferritin ion channels in the water-soluble protein assembly and membrane protein ion channels gated by cytoplasmic N-terminal peptides.

Tosha, Takehiko; Behera, Rabindra K.; Ng, Ho-Leung; Bhattasali, Onita; Alber, Tom; Theil, Elizabeth C.

2012-01-01

11

Effect of Atractylodes macrocephala on Hypertonic Stress-Induced Water Channel Protein Expression in Renal Collecting Duct Cells  

PubMed Central

Edema is a symptom that results from the abnormal accumulation of fluid in the body. The cause of edema is related to the level of aquaporin (AQP)2 protein expression, which regulates the reabsorption of water in the kidney. Edema is caused by overexpression of the AQP2 protein when the concentration of Na+ in the blood increases. The rhizome of Atractylodes macrocephala has been used in traditional oriental medicine as a diuretic drug; however, the mechanism responsible for the diuretic effect of the aqueous extract from A. macrocephala rhizomes (AAMs) has not yet been identified. We examined the effect of the AAM on the regulation of water channels in the mouse inner medullary collecting duct (mIMCD)-3 cells under hypertonic stress. Pretreatment of AAM attenuates a hypertonicity-induced increase in AQP2 expression as well as the trafficking of AQP2 to the apical plasma membrane. Tonicity-responsive enhancer binding protein (TonEBP) is a transcription factor known to play a central role in cellular homeostasis by regulating the expression of some proteins, including AQP2. Western immunoblot analysis demonstrated that the protein and mRNA expression levels of TonEBP also decrease after AAM treatment. These results suggest that the AAM has a diuretic effect by suppressing water reabsorption via the downregulation of the TonEBP-AQP2 signaling pathway.

Lee, Yong Pyo; Lee, Yun Jung; Lee, So Min; Yoon, Jung Joo; Kim, Hye Yoom; Kang, Dae Gill; Lee, Ho Sub

2012-01-01

12

Aquaporins in Saccharomyces: Characterization of a second functional water channel protein.  

PubMed

The Saccharomyces cerevisiae genome database contains two ORFs with homology to aquaporins, AQY1 and AQY2. Aqy1p has been shown to be a functional aquaporin in some strains, such as Sigma1278b. AQY2 is disrupted by a stop codon in most strains; however, Sigma1278b has an intact ORF. Because Sigma1278b Aqy2p has an intracellular localization in Xenopus oocytes and in yeast, other strains of yeast were examined. Aqy2p from Saccharomyces chevalieri has a single amino acid in the third transmembrane domain (Ser-141) that differs from Sigma1278b Aqy2p (Pro-141). S. chevalieri Aqy2p is a functional water channel in oocytes and traffics to the plasma membrane of yeast. The Sigma1278b parental strain, the aqy1-aqy2 double null yeast, and null yeast expressing S. chevalieri Aqy2p were examined under various conditions. Comparison of these strains revealed that the aquaporin null cells were more aggregated and their surface was more hydrophobic. As a result, the aquaporin null cells were more flocculent and more efficient at haploid invasive growth. Despite its primary intracellular localization, Sigma1278b Aqy2p plays a role in yeast similar to Aqy1p and S. chevalieri Aqy2p. In addition, Aqy1p and Aqy2p can affect cell surface properties and may provide an advantage by dispersing the cells during starvation or during sexual reproduction. PMID:11158584

Carbrey, J M; Bonhivers, M; Boeke, J D; Agre, P

2001-01-30

13

Expression of VAMP-2-like protein in kidney collecting duct intracellular vesicles. Colocalization with Aquaporin-2 water channels.  

PubMed Central

Body water balance is controlled by vasopressin, which regulates Aquaporin-2 (AQP2) water channels in kidney collecting duct cells by vesicular trafficking between intracellular vesicles and the plasma membrane. To examine the molecular apparatus involved in vesicle trafficking and vasopressin regulation of AQP2 in collecting duct cells, we tested if targeting proteins expressed in the synaptic vesicles, namely vesicle-associated membrane proteins 1 and 2 (VAMP1 and 2), are expressed in kidney collecting duct. Immunoblotting revealed specific labeling of VAMP2 (18-kD band) but not VAMP1 in membrane fractions prepared from kidney inner medulla. Controls using preadsorbed antibody or preimmune serum were negative. Bands of identical molecular size were detected in immunoblots of brain membrane vesicles and purified synaptic vesicles. VAMP2 in kidney membranes was cleaved by tetanus toxin, revealing a tetanus toxin-sensitive VAMP homologue. Similarly, tetanus toxin cleaved VAMP2 in synaptic vesicles. In kidney inner medulla, VAMP2 was predominantly expressed in the membrane fraction enriched for intracellular vesicles, with little or no VAMP2 in the plasma membrane enriched fraction. This was confirmed by immunocytochemistry using semithin cryosections, which showed mainly vesicular labeling in collecting duct principal cells, with no labeling of intercalated cells. VAMP2 immunolabeling colocalized with AQP2 labeling in intracellular vesicles, as determined by immunoelectron microscopy after double immunolabeling of isolated vesicles. Quantitative analysis of 1,310 vesicles revealed a highly significant association of both AQP2 and VAMP2 in the same vesicles (P < 0.0001). Furthermore, the presence of AQP2 in vesicles immunoisolated with anti-VAMP2 antibodies was confirmed by immunoblotting. In conclusion, VAMP2, a component of the neuronal SNARE complex, is expressed in vesicles carrying AQP2, suggesting a role in vasopressin-regulated vesicle trafficking of AQP2 water channels. Images

Nielsen, S; Marples, D; Birn, H; Mohtashami, M; Dalby, N O; Trimble, M; Knepper, M

1995-01-01

14

Aquaporins: The renal water channels  

PubMed Central

Water is the most abundant molecule in any cell. Specialized membrane channel, proteins called aquaporins, facilitate water transport across cell membranes. At least seven aquaporins (AQP): 1, 2, 3, 4, 6, 7, and 11 are expressed in the kidneys. Aquaporins play a role in both the short-term and long-term regulation of water balance as well as in the pathophysiology of water balance disorders. Aquaporin is composed of a single peptide chain consisting of approximately 270 amino acids. Inherited central and nephrogenic diabetes insipidus are primarily due to the decreased expression of AQP2 while mutation in the AQP2 molecule is responsible for inherited central diabetes insipidus. In acquired causes of nephrogenic diabetes insipidus, there is a downregulation of AQP2 expression in the inner medulla of the kidney. Nephrotic syndrome is characterized by excessive sodium and water reabsorption, although in spite of this, patients do not develop hyponatremia. There is a marked downregulation of both AQP2 and AQP3 expression, which could be a physiologic response to extracellular water reabsorption in patients with nephrotic syndrome. There are some conditions in which aquaporin expression has been found to increase such as experimentally induced heart failure, cirrhosis, and pregnancy. Some drugs such as cisplatin and cyclosporine, also alter the expression of aquaporins. The three-pore model of peritoneal transport depicts the importance of aquaporins. Thus, the understanding of renal water channels has solved the mystery behind many water balance disorders. Further insights into the molecular structure and biology of aquaporins will help to lay a foundation for the development of future drugs.

Agarwal, S. K.; Gupta, A.

2008-01-01

15

FAITH Water Channel Flow Visualization  

NASA Video Gallery

Water channel flow visualization experiments are performed on a three dimensional model of a small hill. This experiment was part of a series of measurements of the complex fluid flow around the hi...

16

Computational optimization of synthetic water channels.  

SciTech Connect

Membranes for liquid and gas separations and ion transport are critical to water purification, osmotic energy generation, fuel cells, batteries, supercapacitors, and catalysis. Often these membranes lack pore uniformity and robustness under operating conditions, which can lead to a decrease in performance. The lack of uniformity means that many pores are non-functional. Traditional membranes overcome these limitations by using thick membrane materials that impede transport and selectivity, which results in decreased performance and increased operating costs. For example, limitations in membrane performance demand high applied pressures to deionize water using reverse osmosis. In contrast, cellular membranes combine high flux and selective transport using membrane-bound protein channels operating at small pressure differences. Pore size and chemistry in the cellular channels is defined uniformly and with sub-nanometer precision through protein folding. The thickness of these cellular membranes is limited to that of the cellular membrane bilayer, about 4 nm thick, which enhances transport. Pores in the cellular membranes are robust under operating conditions in the body. Recent efforts to mimic cellular water channels for efficient water deionization produced a significant advance in membrane function. The novel biomimetic design achieved a 10-fold increase in membrane permeability to water flow compared to commercial membranes and still maintained high salt rejection. Despite this success, there is a lack of understanding about why this membrane performs so well. To address this lack of knowledge, we used highperformance computing to interrogate the structural and chemical environments experienced by water and electrolytes in the newly created biomimetic membranes. We also compared the solvation environments between the biomimetic membrane and cellular water channels. These results will help inform future efforts to optimize and tune the performance of synthetic biomimetic membranes for applications in water purification, energy, and catalysis.

Rogers, David Michael; Rempe, Susan L. B.

2012-12-01

17

Regulation of Ion Channels by G Proteins  

NSDL National Science Digital Library

This Teaching Resource provides lecture notes and slides for a class covering regulation of ion channels by G proteins and is part of the course "Cell Signaling Systems: A Course for Graduate Students." The lecture begins with an overview of calcium channels and then proceeds to describe the interaction of signaling molecules with calcium channels.

Maria Diverse-Pierluissi (Mount Sinai School of Medicine.;Department of Pharmacology and Biological Chemistry REV)

2005-08-16

18

Cotton plasma membrane intrinsic protein 2s (PIP2s) selectively interact to regulate their water channel activities and are required for fibre development.  

PubMed

Aquaporins are thought to be associated with water transport and play important roles in cotton (Gossypium hirsutum) fibre elongation. Among aquaporins, plasma membrane intrinsic proteins (PIPs) constitute a plasma-membrane-specific subfamily and are further subdivided into PIP1 and PIP2 groups. In this study, four fibre-preferential GhPIP2 genes were functionally characterized. The selective interactions among GhPIP2s and their interaction proteins were studied in detail to elucidate the molecular mechanism of cotton fibre development. GhPIP2;3 interacted with GhPIP2;4 and GhPIP2;6, but GhPIP2;6 did not interact with GhPIP2;4. Coexpression of GhPIP2;3/2;4 or GhPIP2;3/2;6 resulted in a positive cooperative effect which increased the permeability coefficient of oocytes, while GhPIP2;4/2;6 did not. GhBCP2 (a blue copper-binding protein) inhibited GhPIP2;6 water channel activity through their interaction. Overexpression of GhPIP2 genes in yeast induced longitudinal growth of the host cells. By contrast, knockdown of expression of GhPIP2 genes in cotton by RNA interference markedly hindered fibre elongation. In conclusion, GhPIP2 proteins are the primary aquaporin isoforms in fibres. They selectively form hetero-oligomers in order to regulate their activities to meet the requirements for rapid fibre elongation. PMID:23656428

Li, Deng-Di; Ruan, Xiang-Mei; Zhang, Jie; Wu, Ya-Jie; Wang, Xiu-Lan; Li, Xue-Bao

2013-08-01

19

Allosteric mechanism of water channel gating by Ca2+-calmodulin  

PubMed Central

Calmodulin (CaM) is a universal regulatory protein that communicates the presence of calcium to its molecular targets and correspondingly modulates their function. This key signaling protein is important for controlling the activity of hundreds of membrane channels and transporters. However, our understanding of the structural mechanisms driving CaM regulation of full-length membrane proteins has remained elusive. In this study, we determined the pseudo-atomic structure of full-length mammalian aquaporin-0 (AQP0, Bos Taurus) in complex with CaM using electron microscopy to understand how this signaling protein modulates water channel function. Molecular dynamics and functional mutation studies reveal how CaM binding inhibits AQP0 water permeability by allosterically closing the cytoplasmic gate of AQP0. Our mechanistic model provides new insight, only possible in the context of the fully assembled channel, into how CaM regulates multimeric channels by facilitating cooperativity between adjacent subunits.

Reichow, Steve L.; Clemens, Daniel M.; Freites, J. Alfredo; Nemeth-Cahalan, Karin L.; Heyden, Matthias; Tobias, Douglas J.; Hall, James E.; Gonen, Tamir

2013-01-01

20

Aquaporin water channels in gastrointestinal physiology  

PubMed Central

Fluid transport is a major function of the gastrointestinal (GI) tract with more than 9 litres of fluid being absorbed or secreted across epithelia in human salivary gland, stomach, the hepatobiliary tract, pancreas, small intestine and colon. This review evaluates the evidence that aquaporin-type water channels are involved in GI fluid transport. The aquaporins are a family of small (?30 kDa) integral membrane proteins that function as water channels. At least seven aquaporins are expressed in various tissues in the GI tract: AQP1 in intrahepatic cholangiocytes, AQP4 in gastric parietal cells, AQP3 and AQP4 in colonic surface epithelium, AQP5 in salivary gland, AQP7 in small intestine, AQP8 in liver, pancreas and colon, and AQP9 in liver. There are functional data suggesting that some GI cell types expressing aquaporins have high or regulated water permeability; however, there has been no direct evidence for a role of aquaporins in GI physiology. Recently, transgenic mice have been generated with selective deletions of various aquaporins. Preliminary evaluation of GI function suggests a role for AQP1 in dietary fat processing and AQP4 in colonic fluid absorption. Further study of aquaporin function in the GI tract should provide new insights into normal GI physiology and disease mechanisms, and may yield novel therapies to regulate fluid movement in GI diseases.

Ma, Tonghui; Verkman, A S

1999-01-01

21

MAL decreases the internalization of the aquaporin-2 water channel  

Microsoft Academic Search

Body water homeostasis depends critically on the hormonally regulated trafficking of aquaporin-2 (AQP2) water channels in renal collecting duct epithelial cells. Several types of posttranslational modifications are clearly involved in controlling the distribution of AQP2 between intracellular vesicles and the apical plasma membrane. Little is known, however, about the protein interactions that govern the trafficking of AQP2 between these organelles.

E.-J. Kamsteeg; A. S. Duffield; I. B. M. Konings; J. Spencer; P. Pagel; P. M. T. Deen; M. J. Caplan

2007-01-01

22

Simulation of Polymer Translocation through Protein Channels.  

National Technical Information Service (NTIS)

A modeling algorithm is presented to compute simultaneously polymer conformations and ionic current, as single polymer molecules undergo translocation through protein channels. The method is based on a combination of Langevin dynamics for coarse-grained m...

M. Muthukumar C. Y. Kong

2005-01-01

23

Ion/water channels for embryo implantation barrier.  

PubMed

Successful implantation involves three distinct processes, namely the embryo apposition, attachment, and penetration through the luminal epithelium of the endometrium to establish a vascular link to the mother. After penetration, stromal cells underlying the epithelium differentiate and surround the embryo to form the embryo implantation barrier, which blocks the passage of harmful substances to the embryo. Many ion/water channel proteins were found to be involved in the process of embryo implantation. First, ion/water channel proteins play their classical role in establishing a resting membrane potential, shaping action potentials and other electrical signals by gating the flow of ions across the cell membrane. Second, most of ion/water channel proteins are regulated by steroid hormone (estrogen or progesterone), which may have important implications to the embryo implantation. Last but not least, these proteins do not limit themselves as pure channels but also function as an initiator of a series of consequences once activated by their ligand/stimulator. Herein, we discuss these new insights in recent years about the contribution of ion/water channels to the embryo implantation barrier construction during early pregnancy. PMID:24789983

Liu, Xin-Mei; Zhang, Dan; Wang, Ting-Ting; Sheng, Jian-Zhong; Huang, He-Feng

2014-05-01

24

Nucleotide sequence and expression of a ripening and water stress-related cDNA from tomato with homology to the MIP class of membrane channel proteins  

Microsoft Academic Search

The nucleotide sequence and derived amino acid sequence were determined for a full-length version of the tomato cDNA clone, pTOM75, the mRNA for which has previously been shown to accumulate in roots, ripening fruit and senescing leaves. Computer analysis of the predicted protein product, which we have named tomato ripening-associated membrane protein (TRAMP) indicates strong homology to known transmembrane channel

Rupert G. Fray; Andrew Wallace; Donald Grierson; Grantley W. Lycett

1994-01-01

25

Theoretical and computational studies of microscopic water channels  

NASA Astrophysics Data System (ADS)

Water channels are ubiquitous in all life forms. A notable example is aquaporins (AQPs), a family of proteins which mainly function as passive water channels in cell membranes. The availability of the crystal structures of several AQPs in recent years allowed us to study them in atomic details. We performed molecular dynamics (MD) simulations on AQPs in lipid bilayers to study water permeation through these channels. We also simulated water conduction in carbon nanotubes, serving as simplified models for biological channels. We developed theories and methodologies aimed to reproduce and predict important experimental quantities of water channels from simulations. We showed that the diffusion permeability (pd), which is measured by tracer diffusion in experiments, can be calculated from equilibrium MD simulations. In order to calculate the osmotic permeability (p f), which is experimentally measured in the presence of a solute concentration difference, we developed a method to induce a hydrostatic pressure difference across the membrane under periodic boundary conditions. We calculated the osmotic permeability for aquaporin-1 using this method, which agrees with experiments. Using a continuous-time random-walk model, we showed that for single-file water channels, the ratio of p f to pd is roughly equal to the number of water molecules in the channel. Proton transfer through single water file was studied theoretically using network thermodynamics. Finally, we proposed a new model for general water channels, which gives a quantitative relationship between water permeations under equilibrium and non-equilibrium conditions, and therefore allows one to calculate pf from equilibrium MD simulations.

Zhu, Fangqiang

26

Rapid stalk elongation in tulip (Tulipa gesneriana L. cv. Apeldoorn) and the combined action of cold-induced invertase and the water-channel protein gammaTIP.  

PubMed

Many bulbous plants need a low-temperature treatment for flowering. Cold, for example, affects the elongation of the stalk, thereby influencing the quality of the cut flower. How the elongation of the stalk is promoted by cold and which physiological and biochemical mechanisms are involved have remained obscure. As invertase has been shown to be involved in the cold-induced elongation of the flower stalks of tulips (Lambrechts et al., 1994, Plant Physiol 104: 515-520), we further characterized this enzyme by cloning the cDNA and analysing its expression in various tissues of the tulip (Tulipa gesneriana L. cv. Apeldoorn) stalk. In addition, the role of sucrose synthase was investigated. Since turgor pressure is an important force driving cell elongation, the role of a water-channel protein (gammaTIP) was studied in relation to these two enzymes. The mRNA level of the invertase found was substantially up-regulated as a result of cold treatment. Analysis of the amino acid sequence of this invertase revealed the presence of a vacuolar targeting signal. Two different forms of sucrose synthase were found, the expression of one of them appeared to be restricted to the vascular tissue while the other form was present in the surrounding tissue. Both sucrose synthases were present in the stalk during the entire period of bulb storage and after planting, but their activities declined during stalk elongation. The expression of the gammaTIP gene was restricted mainly to the vascular tissue and its expression profile was identical to that of invertase. Simultaneous expression of invertase and gammaTIP possibly leads to an increase in osmotic potential and vacuolar water uptake, thus providing a driving force for stretching the stalk cells. PMID:10502102

Balk, P A; de Boer, A D

1999-09-01

27

Phylogenetic Characterization of the MIP Family of Transmembrane Channel Proteins  

Microsoft Academic Search

.   The ubiquitous major intrinsic protein (MIP) family includes several transmembrane channel proteins known to exhibit specificity\\u000a for water and\\/or neutral solutes. We have identified 84 fully or partially sequenced members of this family, have multiply\\u000a aligned over 50 representative, divergent, fully sequenced members, have used the resultant multiple alignment to derive current\\u000a MIP family-specific signature sequences, and have constructed

J. H. Park

1996-01-01

28

A SUBSTRATE CHANNEL IN THE NITROGENASE MoFe PROTEIN  

PubMed Central

Nitrogenase catalyzes the six electron/six proton reduction of N2 to two ammonia molecules at a complex organo-metallocluster called FeMo-cofactor. This cofactor is buried within the ?-subunit of the MoFe protein, with no obvious access for substrates. Examination of high-resolution X-ray crystal structures of MoFe proteins from several organisms has revealed the existence of a water-filled channel that extends from the solvent exposed surface to a specific face of FeMo-cofactor. This channel could provide a pathway for substrate and product access to the active site. In the present work, we examine this possibility by substituting four different amino acids that line the channel with other residues and analyze the impact of these substitutions on substrate reduction kinetic parameters. Each of the MoFe protein variants was purified and kinetic parameters established for the reduction of the substrates N2, acetylene, azide and propyne. For each MoFe protein, Vmax values for the different substrates were found to be nearly unchanged when compared to the wild-type MoFe protein, indicating that electron delivery to the active site is not compromised by the various substitutions. In contrast, the Km values for these substrates were found to increase significantly (up to 22-fold) in some of the MoFe protein variants compared to the wild-type MoFe protein values. Given that each of the amino acids that were substituted is remote from the active site, these results are consistent with the water filled channel functioning as a substrate channel in the MoFe protein.

Barney, Brett M.; Yurth, Michael G.; Dos Santos, Patricia C.; Dean, Dennis R.; Seefeldt, Lance C.

2010-01-01

29

Reconstitution of a chloroplast protein import channel.  

PubMed Central

The chloroplastic outer envelope protein OEP75 with a molecular weight of 75 kDa probably forms the central pore of the protein import machinery of the outer chloroplastic membrane. Patch-clamp analysis shows that heterologously expressed, purified and reconstituted OEP75 constitutes a voltage-gated ion channel with a unit conductance of Lambda = 145pS. Activation of the OEP75 channel in vitro is completely dependent on the magnitude and direction of the voltage gradient. Therefore, movements of protein charges of parts of OEP75 in the membrane electric field are required either for pore formation or its opening. In the presence of precursor protein from only one side of the bilayer, strong flickering and partial closing of the channel was observed, indicating a specific interaction of the precursor with OEP75. The comparatively low ionic conductance of OEP75 is compatible with a rather narrow aqueous pore (dporeapproximately equal to 8-9 A). Provided that protein and ion translocation occur through the same pore, this implies that the environment of the polypeptide during the transit is mainly hydrophilic and that protein translocation requires almost complete unfolding of the precursor.

Hinnah, S C; Hill, K; Wagner, R; Schlicher, T; Soll, J

1997-01-01

30

Origin of Martian channels - Clathrates and water  

NASA Technical Reports Server (NTRS)

Criticism is directed at the suggestion that Martian channels may have been eroded by liquid water produced by the depressurization of CO2 hydrate. The release of pre-existing subsurface liquid water, such as that trapped under a permafrost layer, by meteorite impact or tectonic activity could produce sufficient flow and would not require heat transfer. The presence of water in a CO2 hydrate is shown to be detrimental to its release from an underground reservoir.

Peale, S. J.; Schubert, G.; Lingenfelter, R. E.

1975-01-01

31

Desformylgramicidin: a model channel with an extremely high water permeability.  

PubMed Central

The water conductivity of desformylgramicidin exceeds the permeability of gramicidin A by two orders of magnitude. With respect to its single channel hydraulic permeability coefficient of 1.1.10(-12) cm(3) s(-1), desformylgramicidin may serve as a model for extremely permeable aquaporin water channel proteins (AQP4 and AQPZ). This osmotic permeability exceeds the conductivity that is predicted by the theory of single-file transport. It was derived from the concentration distributions of both pore-impermeable and -permeable cations that were simultaneously measured by double barreled microelectrodes in the immediate vicinity of a planar bilayer. From solvent drag experiments, approximately five water molecules were found to be transported by a single-file process along with one ion through the channel. The single channel proton, potassium, and sodium conductivities were determined to be equal to 17 pS (pH 2.5), 7 and 3 pS, respectively. Under any conditions, the desformyl-channel remains at least 10 times longer in its open state than gramicidin A.

Saparov, S M; Antonenko, Y N; Koeppe, R E; Pohl, P

2000-01-01

32

1. INTAKE CHANNEL LOOKING NORTHEAST; WATER FROM BEAVER BROOK ENTERS ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

1. INTAKE CHANNEL LOOKING NORTHEAST; WATER FROM BEAVER BROOK ENTERS THE INTAKE CHANNEL HERE. - Hondius Water Line, 1.6 miles Northwest of Park headquarters building & 1 mile Northwest of Beaver Meadows entrance station, Estes Park, Larimer County, CO

33

Simulation of polymer translocation through protein channels  

NASA Astrophysics Data System (ADS)

A modeling algorithm is presented to compute simultaneously polymer conformations and ionic current, as single polymer molecules undergo translocation through protein channels. The method is based on a combination of Langevin dynamics for coarse-grained models of polymers and the Poisson-Nernst-Planck formalism for ionic current. For the illustrative example of ssDNA passing through the -hemolysin pore, vivid details of conformational fluctuations of the polymer inside the vestibule and -barrel compartments of the protein pore, and their consequent effects on the translocation time and extent of blocked ionic current are presented. In addition to yielding insights into several experimentally reported puzzles, our simulations offer experimental strategies to sequence polymers more efficiently.

Muthukumar, M.; Kong, C. Y.

2006-04-01

34

The enigma of the CLIC proteins: Ion channels, redox proteins, enzymes, scaffolding proteins?  

Microsoft Academic Search

Chloride intracellular channel proteins (CLICs) are distinct from most ion channels in that they have both soluble and integral membrane forms. CLICs are highly conserved in chordates, with six vertebrate paralogues. CLIC-like proteins are found in other metazoans. CLICs form channels in artificial bilayers in a process favoured by oxidising conditions and low pH. They are structurally plastic, with CLIC1

Dene R. Littler; Stephen J. Harrop; Sophia C. Goodchild; Juanita M. Phang; Andrew V. Mynott; Lele Jiang; Stella M. Valenzuela; Michele Mazzanti; Louise J. Brown; Samuel N. Breit; Paul M. G. Curmi

2010-01-01

35

Lipid ion channels and the role of proteins.  

PubMed

In the absence of proteins, synthetic lipid membranes can display quantized conduction events for ions that are virtually indistinguishable from those of protein channels. The phenomenological similarities between typical conductances are striking: they are of equal order and show similar lifetime distributions and current histograms. They can include conduction bursts, flickering, and multistep conductance. Lipid channels can be gated by voltage and blocked by drugs. They respond to changes in lateral membrane tension and temperature. Thus, they behave like voltage-gated, temperature-gated, and mechano-sensitive protein channels, or like receptors. The similarity between lipid and protein channels poses an important problem for the interpretation of protein channel data. For example, the Hodgkin-Huxley theory for nerve pulse conduction requires a selective mechanism for the conduction of sodium and potassium ions. To this end, the lipid membrane must act both as a capacitor and as an insulator. Nonselective ion conductance by mechanisms other than the gated protein channels challenges the proposed mechanism for pulse propagation. Nevertheless, textbooks rarely describe the properties of the lipid membrane surrounding the proteins in their discussions of membrane models. These similarities lead to important questions: Do these similarities in lipid and protein channels result from a common mechanism, or are these similarities fortuitous? What distinguishes protein channels from lipid channels, if anything? In this Account, we document experimental and theoretical findings that show the similarity between lipid and protein channels. We discuss important cases where protein channel function strongly correlates with the properties of the lipid. Based on statistical thermodynamics simulations, we discuss how such correlations could come about. We suggest that proteins can act as catalysts for lipid channel formation and that this hypothesis can explain some of the unexplained correlations between protein and lipid membrane function. PMID:23902303

Mosgaard, Lars D; Heimburg, Thomas

2013-12-17

36

Aquaporin water channels in the nervous system  

PubMed Central

The aquaporins (AQPs) are plasma membrane water-transporting proteins. AQP4 is the principal member of this protein family in the CNS, where it is expressed in astrocytes and is involved in water movement, cell migration and neuroexcitation. AQP1 is expressed in the choroid plexus, where it facilitates cerebrospinal fluid secretion, and in dorsal root ganglion neurons, where it tunes pain perception. The AQPs are potential drug targets for several neurological conditions. Astrocytoma cells strongly express AQP4, which may facilitate their infiltration into the brain, and the neuroinflammatory disease neuromyelitis optica is caused by AQP4-specific autoantibodies that produce complement-mediated astrocytic damage.

Papadopoulos, Marios C.; Verkman, Alan S.

2013-01-01

37

Biomphalaria species in Alexandria water channels.  

PubMed

Of the several species of Biomphalaria snails worldwide that serve as the intermediate host for Schistosoma mansoni, Biomphalaria alexandrina is a species that is indigenous to Egypt. Recently, there has been much debate concerning the presence of Biomphalaria glabrata and the hybrid of the species with Biomphalaria alexandrina. Due to this debate, the absence of a clear explanation for the presence of B. glabrata in Egyptian water channels and the probability that they may be reintroduced, we conducted this field study to identify Biomphalaria species present in Alexandria water channels. Laboratory-adapted susceptible snails to Schistosoma mansoni of the following species were used as a reference; Biomphalaria alexandrina, Biomphalaria glabrata and their hybrid. These snails were used to perpetuate the Schistosoma life cycle at the Theodor Bilharz Research Institute (TBRI), Cairo, Egypt. Morphological and molecular studies were conducted on these reference snails as well as on the first generation of Biomphalaria snails from two areas in the Alexandria governorate. The morphological study included both external shell morphology and internal anatomy of the renal ridge. The molecular study used a species-specific PCR technique. The results demonstrated that there was an absence of Biomphalaria glabrata and the hybrid from Alexandria water channels. Moreover, the susceptibility patterns of these reference snails were studied by measuring the different parasitological parameters. It was found that Biomphalaria glabrata and the hybrid were significantly more susceptible than Biomphalaria alexandrina to the Egyptian strain of Schistosoma mansoni. The results demonstrated that if Biomphalaria glabrata was reintroduced and adapted to the local environment in Egypt, it would have important epidemiologic impacts that would have a serious effect on the health of Egyptian people. PMID:21458594

Abou-El-Naga, Iman F; El-Nassery, Suzanne M F; Allam, Sonia R; Shaat, Eman A; Mady, Rasha F M

2011-09-01

38

Ion channels induced by the prion protein  

PubMed Central

Prion diseases comprise a group of rapidly progressive and invariably fatal neurodegenerative disorders for which there are no effective treatments. While conversion of the cellular prion protein (PrPC) to a ?-sheet rich isoform (PrPSc) is known to be a critical event in propagation of infectious prions, the identity of the neurotoxic form of PrP and its mechanism of action remain unclear. Insights into this mechanism have been provided by studying PrP molecules harboring deletions and point mutations in the conserved central region, encompassing residues 105–125. When expressed in transgenic mice, PrP deleted for these residues (?105–125) causes a spontaneous neurodegenerative illness that is reversed by co-expression of wild-type PrP. In cultured cells, ?105–125 PrP confers hypersensitivity to certain cationic antibiotics and induces spontaneous ion channel activity that can be recorded by electrophysiological techniques. We have utilized these drug-hypersensitization and current-inducing activities to identify which PrP domains and subcellular locations are required for toxicity. We present an ion channel model for the toxicity of ?105–125 PrP and related mutants and speculate how a similar mechanism could mediate PrPSc-associated toxicity. Therapeutic regimens designed to inhibit prion-induced toxicity, as well as formation of PrPSc, may prove to be the most clinically beneficial.

Solomon, Isaac H; Biasini, Emiliano

2012-01-01

39

A kinetic role for ionizable sites in membrane channel proteins  

Microsoft Academic Search

Electrically charged residues in a membrane channel protein will certainly have a direct effect upon its gating and selectivity if they are near the channel pore. It is customary to regard the charged state of such residues as a fixed feature of the channel. In this paper it is argued that far from being fixed, the charged state of ionizable

D. T. Edmonds

1989-01-01

40

GIRK channel modulation by assembly with allosterically regulated RGS proteins  

PubMed Central

G-protein–activated inward-rectifying K+ (GIRK) channels hyperpolarize neurons to inhibit synaptic transmission throughout the nervous system. By accelerating G-protein deactivation kinetics, the regulator of G-protein signaling (RGS) protein family modulates the timing of GIRK activity. Despite many investigations, whether RGS proteins modulate GIRK activity in neurons by mechanisms involving kinetic coupling, collision coupling, or macromolecular complex formation has remained unknown. Here we show that GIRK modulation occurs by channel assembly with R7-RGS/G?5 complexes under allosteric control of R7 RGS-binding protein (R7BP). Elimination of R7BP occludes the G?5 subunit that interacts with GIRK channels. R7BP-bound R7-RGS/G?5 complexes and G?? dimers interact noncompetitively with the intracellular domain of GIRK channels to facilitate rapid activation and deactivation of GIRK currents. By disrupting this allosterically regulated assembly mechanism, R7BP ablation augments GIRK activity. This enhanced GIRK activity increases the drug effects of agonists acting at G-protein–coupled receptors that signal via GIRK channels, as indicated by greater antinociceptive effects of GABA(B) or ?-opioid receptor agonists. These findings show that GIRK current modulation in vivo requires channel assembly with allosterically regulated RGS protein complexes, which provide a target for modulating GIRK activity in neurological disorders in which these channels have crucial roles, including pain, epilepsy, Parkinson’s disease and Down syndrome.

Zhou, Hao; Chisari, Mariangela; Raehal, Kirsten M.; Kaltenbronn, Kevin M.; Bohn, Laura M.; Mennerick, Steven J.; Blumer, Kendall J.

2012-01-01

41

GIRK channel modulation by assembly with allosterically regulated RGS proteins.  

PubMed

G-protein-activated inward-rectifying K(+) (GIRK) channels hyperpolarize neurons to inhibit synaptic transmission throughout the nervous system. By accelerating G-protein deactivation kinetics, the regulator of G-protein signaling (RGS) protein family modulates the timing of GIRK activity. Despite many investigations, whether RGS proteins modulate GIRK activity in neurons by mechanisms involving kinetic coupling, collision coupling, or macromolecular complex formation has remained unknown. Here we show that GIRK modulation occurs by channel assembly with R7-RGS/G?5 complexes under allosteric control of R7 RGS-binding protein (R7BP). Elimination of R7BP occludes the G?5 subunit that interacts with GIRK channels. R7BP-bound R7-RGS/G?5 complexes and G?? dimers interact noncompetitively with the intracellular domain of GIRK channels to facilitate rapid activation and deactivation of GIRK currents. By disrupting this allosterically regulated assembly mechanism, R7BP ablation augments GIRK activity. This enhanced GIRK activity increases the drug effects of agonists acting at G-protein-coupled receptors that signal via GIRK channels, as indicated by greater antinociceptive effects of GABA(B) or ?-opioid receptor agonists. These findings show that GIRK current modulation in vivo requires channel assembly with allosterically regulated RGS protein complexes, which provide a target for modulating GIRK activity in neurological disorders in which these channels have crucial roles, including pain, epilepsy, Parkinson's disease and Down syndrome. PMID:23169654

Zhou, Hao; Chisari, Mariangela; Raehal, Kirsten M; Kaltenbronn, Kevin M; Bohn, Laura M; Mennerick, Steven J; Blumer, Kendall J

2012-12-01

42

The enigma of the CLIC proteins: Ion channels, redox proteins, enzymes, scaffolding proteins?  

PubMed

Chloride intracellular channel proteins (CLICs) are distinct from most ion channels in that they have both soluble and integral membrane forms. CLICs are highly conserved in chordates, with six vertebrate paralogues. CLIC-like proteins are found in other metazoans. CLICs form channels in artificial bilayers in a process favoured by oxidising conditions and low pH. They are structurally plastic, with CLIC1 adopting two distinct soluble conformations. Phylogenetic and structural data indicate that CLICs are likely to have enzymatic function. The physiological role of CLICs appears to be maintenance of intracellular membranes, which is associated with tubulogenesis but may involve other substructures. PMID:20085760

Littler, Dene R; Harrop, Stephen J; Goodchild, Sophia C; Phang, Juanita M; Mynott, Andrew V; Jiang, Lele; Valenzuela, Stella M; Mazzanti, Michele; Brown, Louise J; Breit, Samuel N; Curmi, Paul M G

2010-05-17

43

Putting proteins back into water  

NASA Astrophysics Data System (ADS)

We introduce a simplified protein model where the solvent (water) degrees of freedom appear explicitly (although in an extremely simplified fashion). Using this model we are able to recover the thermodynamic phenomenology of proteins over a wide range of temperatures. In particular we describe both the warm and the cold protein denaturation within a single framework, while addressing important issues about the structure of model proteins.

de Los Rios, Paolo; Caldarelli, Guido

2000-12-01

44

Mechanosensitivity of ion channels based on protein-lipid interactions  

PubMed Central

Ion channels form a group of membrane proteins that pass ions through a pore beyond the energy barrier of the lipid bilayer. The structure of the transmembrane segment of membrane proteins is influenced by the charges and the hydrophobicity of the surrounding lipids and the pressure on its surface. A mechanosensitive channel is specifically designed to change its conformation in response to changes in the membrane pressure (tension). However, mechanosensitive channels are not the only group that is sensitive to the physical environment of the membrane: voltage-gated channels are also amenable to the lipid environment. In this article, we review the structure and gating mechanisms of the mechanosensitive channels and voltage-gated channels and discuss how their functions are affected by the physical properties of the lipid bilayer.

Yoshimura, Kenjiro; Sokabe, Masahiro

2010-01-01

45

BIOCHEMISTRY: TRP Ion Channels--Two Proteins in One  

NSDL National Science Digital Library

Access to the article is free, however registration and sign-in are required. It is well established that some proteins carry out more than one job in the cell, but so far, ion channels do not appear to exhibit this versatility. In their Perspective, Levitan and Cibulsky discuss a cluster of new findings showing that two members of the long TRP ion channel family, LTRPC7 and LTRPC2, are both ion channels and enzymes.

Irwin B. Levitan (University of Pennsylvania School of Medicine;Department of Neuroscience); Susan M. Cibulsky (University of Pennsylvania School of Medicine;Department of Neuroscience)

2001-08-17

46

Activation of maxi-anion channel by protein tyrosine dephosphorylation.  

PubMed

The maxi-anion channel with a large single-channel conductance of >300 pS, and unknown molecular identity, is functionally expressed in a large variety of cell types. The channel is activated by a number of experimental maneuvers such as exposing cells to hypotonic or ischemic stress. The most effective and consistent method of activating it is patch membrane excision. However, the activation mechanism of the maxi-anion channel remains poorly understood at present. In the present study, involvement of phosphorylation/dephosphorylation in excision-induced activation was examined. In mouse mammary fibroblastic C127 cells, activity of the channel was suppressed by intracellular application of Mg-ATP, but not Mg-5'-adenylylimidodiphosphate (AMP-PNP), in a concentration-dependent manner. When a cocktail of broad-spectrum tyrosine phosphatase inhibitors was applied, channel activation was completely abolished, whereas inhibitors of serine/threonine protein phosphatases had no effect. On the other hand, protein tyrosine kinase inhibitors brought the channel out of an inactivated state. In mouse adult skin fibroblasts (MAFs) in primary culture, similar maxi-anion channels were found to be activated on membrane excision, in a manner sensitive to tyrosine phosphatase inhibitors. In MAFs isolated from animals deficient in receptor protein tyrosine phosphatase (RPTP)zeta, activation of the maxi-anion channel was significantly slower and less prominent compared with that observed in wild-type MAFs; however, channel activation was restored by transfection of the RPTPzeta gene. Thus it is concluded that activation of the maxi-anion channel involves protein dephosphorylation mediated by protein tyrosine phosphatases that include RPTPzeta in mouse fibroblasts, but not in C127 cells. PMID:19657061

Toychiev, Abduqodir H; Sabirov, Ravshan Z; Takahashi, Nobuyaki; Ando-Akatsuka, Yuhko; Liu, Hongtao; Shintani, Takafumi; Noda, Masaharu; Okada, Yasunobu

2009-10-01

47

Calcium channel ? subunits: a functionally diverse protein family  

Microsoft Academic Search

The calcium channel ? subunits comprise an eight-member protein family that share a common topology consisting of four transmembrane\\u000a domains and intracellular N- and C-termini. Although the first ? subunit was identified as an auxiliary subunit of a voltage-dependent\\u000a calcium channel, a review of phylogenetic, bioinformatic, and functional studies indicates that they are a functionally diverse\\u000a protein family. A cluster

Ren-Shiang Chen; Tzyy-Chyn Deng; Thomas Garcia; Zachary M. Sellers; Philip M. Best

2007-01-01

48

Identification of chloride intracellular channel proteins in spermatozoa  

Microsoft Academic Search

We have identified for the first time the presence of chloride intracellular channel (CLIC) proteins in bovine epididymal spermatozoa. CLIC1 was discovered during microsequencing of proteins that co-purified with protein phosphatase 1, PP1?2, in sperm extracts. In addition to CLIC1, Western blot showed that two additional CLIC family members, CLIC4 and CLIC5, are also present in spermatozoa. CLIC fusion proteins,

Kimberley Myers; Payaningal R. Somanath; Mark Berryman; Srinivasan Vijayaraghavan

2004-01-01

49

Interaction of Sedlin with chloride intracellular channel proteins  

Microsoft Academic Search

Sedlin is an evolutionarily conserved protein encoded by the causative gene SEDL for spondyloepiphyseal dysplasia tarda. Nevertheless, how Sedlin mutations cause the disease remains unknown. Here, the intracellular chloride channel protein CLIC1 was shown to associate with Sedlin by yeast two-hybrid screening. Green fluorescence protein-CLIC1 readily co-immunoprecipitated with FLAG-Sedlin. In addition, both proteins colocalized extensively in cytoplasmic vesicular\\/reticular structures in

Libin Fan; Wei Yu; Xueliang Zhu

2003-01-01

50

Aram Chaos outflow channel: water volume and time scale  

NASA Astrophysics Data System (ADS)

The evaluation of the water volume and the formative time scale needed to carve the outflow channels represents a fundamental process for the validation of their evolutive models. We calculate these attributes for the Aram channels and we compared the results with the volume of liquid water that was produced in a single chaotization event of the Aram Chaos. The analysis suggests that a single rapid and catastrophic event is sufficient to carve the channel and the volume of flood is compatible with the volume of liquid water release in a single chaotization event of the Aram Chaos.

Roda, M.; Kleinhans, M. G.; Zegers, T. E.

2012-09-01

51

[VC and DCE in groundwater and drainage channel water].  

PubMed

In an area used merely for gardening in a downland moor, which is partly transformed to an industrial estate, accidentally a contamination of a drainage channel with VOC's - predominantly chloroethylene (vinyl chloride [VC]) and 1.2-cis-dichloroethylene (DCE) - was found. The ascending ground water leaks into the drainage channels. The dissolved harmful substances (water solubility of VC is 1.6 g/l) can reach the radix range of plants and fruit bosks and can theoretically be incorporated with the water influx. Additionally the water from the drainage channels can be used to water the crops. Six gardens and a housing were involved. In the groundwater of the mainly concerned region max. 5,000 microg/l VOC's (quite predominantly VC and DCE) was measured from 147 samples. In the drainage channel water max. 2,500 microg/l was measured from 52 samples (limit value according to the drinking water ordinance is 10 microg/l). In the sediment of the channel with approximately 60,000 microg/kg VOC was found in dry matter (6 samples). We describe, how the consumer protection dept. dealt with this unexpected situation and what measures were taken. The impact on human health by the contaminated ground and channel water or by means of contaminated plants are determined for tree fruits, potatoes, bulbs and carrots. The soil air was contaminated, but in buildings no harmful compounds were detectioned. PMID:15609218

Ackermann, A

2004-12-01

52

Anoctamin and transmembrane channel-like proteins are evolutionarily related  

PubMed Central

Anoctamin (ANO) family of proteins, consisting of 10 members in mammals, are transmembrane proteins that have Ca2+-activated Cl- channel activities. Transmembrane channel-like (TMC) family of proteins, consisting of 8 members in mammals, are also transmembrane proteins of which mutations are implicated in various human conditions, such as hearing loss and epidermodysplasia verruciformis. Here we show that ANO and TMC proteins share high sequence similarity and probably the same membrane topology, indicating that these proteins are evolutionarily related. We found many conserved amino acid residues between the two families of proteins, especially in regions spanning the transmembrane domains TM1, TM4-TM5, and TM6-TM7. These findings imply that these proteins form one large family, which we term ANO/TMC superfamily and that TMC proteins may also function as channels for Cl- or possibly other ions. The ANO/TMC superfamily proteins are present in almost all the diverse groups of eukaryotic organisms, suggesting that the proteins function in important biological processes, such as ion homeostasis, in eukaryotic cells.

HAHN, YOONSOO; KIM, DONG SEON; PASTAN, IRA H.; LEE, BYUNGKOOK

2009-01-01

53

WATER TEMPERATURE DYNAMICS IN EXPERIMENTAL FIELD CHANNELS: ANALYSIS AND MODELING  

EPA Science Inventory

This study is on water temperature dynamics in the shallow field channels of the USEPA Monticello Ecological Research Station (MERS). The hydraulic and temperature environment in the MERS channels was measured and simulated to provide some background for several biological studie...

54

Impaired olfaction in mice lacking aquaporin-4 water channels  

PubMed Central

Aquaporin-4 (AQP4) is a water-selective transport protein expressed in glial cells throughout the central nervous system. AQP4 deletion in mice produces alterations in several neuroexcitation phenomena, including hearing, vision, epilepsy, and cortical spreading depression. Here, we report defective olfaction and electroolfactogram responses in AQP4-null mice. Immunofluorescence indicated strong AQP4 expression in supportive cells of the nasal olfactory epithelium. The olfactory epithelium in AQP4-null mice had identical appearance, but did not express AQP4, and had ?12-fold reduced osmotic water permeability. Behavioral analysis showed greatly impaired olfaction in AQP4-null mice, with latency times of 17 ± 0.7 vs. 55 ± 5 s in wild-type vs. AQP4-null mice in a buried food pellet test, which was confirmed using an olfactory maze test. Electroolfactogram voltage responses to multiple odorants were reduced in AQP4-null mice, with maximal responses to triethylamine of 0.80 ± 0.07 vs. 0.28 ± 0.03 mV. Similar olfaction and electroolfactogram defects were found in outbred (CD1) and inbred (C57/bl6) mouse genetic backgrounds. Our results establish AQP4 as a novel determinant of olfaction, the deficiency of which probably impairs extracellular space K+ buffering in the olfactory epithelium.—Lu, D. C., Zhang, H., Zador, Z., Verkman, A. S. Impaired olfaction in mice lacking aquaporin-4 water channels.

Lu, Daniel C.; Zhang, Hua; Zador, Zsolt; Verkman, A. S.

2008-01-01

55

Water quality control in open channels  

Microsoft Academic Search

With the increasing degree of automation in Dutch water management the need for control systems and design procedures for control systems is also evolving. Traditionally these control systems were used for water level and discharge control only. With the measurement equipment that is currently available, improved water quality control also becomes possible. A PhD researcher is currently investigating the theoretical

A. Hof; W. Schuurmans

56

Vegetation and Channel Morphology Responses to Ordinary High Water Discharge Events in Arid West Stream Channels.  

National Technical Information Service (NTIS)

Waters of the United States (WoUS) are regulated by the U.S. Army Corps of Engineers under Section 404 of the Clean Water Act (33 U.S.C. 1344). The Corps lateral jurisdictional extent in Arid West stream channels is the upper level of the ordinary high wa...

B. Allen C. Photos D. Cate L. Dixon R. Lichvar

2009-01-01

57

Model studies of dense water overflows in the Faroese Channels  

NASA Astrophysics Data System (ADS)

The overflow of dense water from the Nordic Seas through the Faroese Channel system was investigated through combined laboratory experiments and numerical simulations using the Massachusetts Institute of Technology General Circulation Model. In the experimental study, a scaled, topographic representation of the Faroe-Shetland Channel, Wyville-Thomson Basin and Ridge and Faroe Bank Channel seabed bathymetry was constructed and mounted in a rotating tank. A series of parametric experiments was conducted using dye-tracing and drogue-tracking techniques to investigate deep-water overflow pathways and circulation patterns within the modelled region. In addition, the structure of the outflowing dense bottom water was investigated through density profiling along three cross-channel transects located in the Wyville-Thomson Basin and the converging, up-sloping approach to the Faroe Bank Channel. Results from the dye-tracing studies demonstrate a range of parametric conditions under which dense water overflow across the Wyville-Thomson Ridge is shown to occur, as defined by the Burger number, a non-dimensional length ratio and a dimensionless dense water volume flux parameter specified at the Faroe-Shetland Channel inlet boundary. Drogue-tracking measurements reveal the complex nature of flow paths and circulations generated in the modelled topography, particularly the development of a large anti-cyclonic gyre in the Wyville-Thompson Basin and up-sloping approach to the Faroe Bank Channel, which diverts the dense water outflow from the Faroese shelf towards the Wyville-Thomson Ridge, potentially promoting dense water spillage across the ridge itself. The presence of this circulation is also indicated by associated undulations in density isopycnals across the Wyville-Thomson Basin. Numerical simulations of parametric test cases for the main outflow pathways and density structure in a similarly-scaled Faroese Channels model domain indicate excellent qualitative agreement with the experimental observations and measurements. In addition, the comparisons show that strong temporal variability in the predicted outflow pathways and circulations have a strong influence in regulating the Faroe Bank Channel and Wyville-Thomson Ridge overflows, as well as in determining the overall response in the Faroese Channels to changes in the Faroe-Shetland Channel inlet boundary conditions.

Cuthbertson, Alan; Davies, Peter; Stashchuk, Nataliya; Vlasenko, Vasiliy

2014-01-01

58

Structural basis of water-specific transport through the AQP1 water channel  

NASA Astrophysics Data System (ADS)

Water channels facilitate the rapid transport of water across cell membranes in response to osmotic gradients. These channels are believed to be involved in many physiological processes that include renal water conservation, neuro-homeostasis, digestion, regulation of body temperature and reproduction. Members of the water channel superfamily have been found in a range of cell types from bacteria to human. In mammals, there are currently 10 families of water channels, referred to as aquaporins (AQP): AQP0-AQP9. Here we report the structure of the aquaporin 1 (AQP1) water channel to 2.2Å resolution. The channel consists of three topological elements, an extracellular and a cytoplasmic vestibule connected by an extended narrow pore or selectivity filter. Within the selectivity filter, four bound waters are localized along three hydrophilic nodes, which punctuate an otherwise extremely hydrophobic pore segment. This unusual combination of a long hydrophobic pore and a minimal number of solute binding sites facilitates rapid water transport. Residues of the constriction region, in particular histidine 182, which is conserved among all known water-specific channels, are critical in establishing water specificity. Our analysis of the AQP1 pore also indicates that the transport of protons through this channel is highly energetically unfavourable.

Sui, Haixin; Han, Bong-Gyoon; Lee, John K.; Walian, Peter; Jap, Bing K.

2001-12-01

59

5. GATE 5, INTAKE CHANNEL LOOKING SOUTH; WATER FROM GATE ...  

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

5. GATE 5, INTAKE CHANNEL LOOKING SOUTH; WATER FROM GATE 5 ENTERED DITCH AND IRRIGATED HONDIUS' FIELDS. - Hondius Water Line, 1.6 miles Northwest of Park headquarters building & 1 mile Northwest of Beaver Meadows entrance station, Estes Park, Larimer County, CO

60

Interaction of Sedlin with chloride intracellular channel proteins.  

PubMed

Sedlin is an evolutionarily conserved protein encoded by the causative gene SEDL for spondyloepiphyseal dysplasia tarda. Nevertheless, how Sedlin mutations cause the disease remains unknown. Here, the intracellular chloride channel protein CLIC1 was shown to associate with Sedlin by yeast two-hybrid screening. Green fluorescence protein-CLIC1 readily co-immunoprecipitated with FLAG-Sedlin. In addition, both proteins colocalized extensively in cytoplasmic vesicular/reticular structures in COS-7 cells, suggesting their interaction at intracellular membranous organelles. Sedlin also associated with CLIC2 in yeast two-hybrid assays. The link between Sedlin and the intracellular chloride channels is the first step to understand their functional interplays. PMID:12681486

Fan, Libin; Yu, Wei; Zhu, Xueliang

2003-04-10

61

Aquaporin-1 channel function is positively regulated by protein kinase C.  

PubMed

Aquaporin-1 (AQP1) channels contribute to osmotically induced water transport in several organs including the kidney and serosal membranes such as the peritoneum and the pleura. In addition, AQP1 channels have been shown to conduct cationic currents upon stimulation by cyclic nucleotides. To date, the short term regulation of AQP1 function by other major intracellular signaling pathways has not been studied. In the present study, we therefore investigated the regulation of AQP1 by protein kinase C. AQP1 wild type channels were expressed in Xenopus oocytes. Water permeability was assessed by hypotonic challenges. Activation of protein kinase C (PKC) by 1-oleoyl-2-acetyl-sn-glycerol (OAG) induced a marked increase of AQP1-dependent water permeability. This regulation was abolished in mutated AQP1 channels lacking both consensus PKC phosphorylation sites Thr(157) and Thr(239) (termed AQP1 DeltaPKC). AQP1 cationic currents measured with double-electrode voltage clamp were markedly increased after pharmacological activation of PKC by either OAG or phorbol 12-myristate 13-acetate. Deletion of either Thr(157) or Thr(239) caused a marked attenuation of PKC-dependent current increases, and deletion of both phosphorylation sites in AQP1 DeltaPKC channels abolished the effect. In vitro phosphorylation studies with synthesized peptides corresponding to amino acids 154-168 and 236-250 revealed that both Thr(157) and Thr(239) are phosphorylated by PKC. Upon stimulation by cyclic nucleotides, AQP1 wild type currents exhibited a strong activation. This regulation was not affected after deletion of PKC phosphorylation sites in AQP1 DeltaPKC channels. In conclusion, this is the first study to show that PKC positively regulates both water permeability and ionic conductance of AQP1 channels. This new pathway of AQP1 regulation is independent of the previously described cyclic nucleotide pathway and may contribute to the PKC stimulation of AQP1-modulated processes such as endothelial permeability, angiogenesis, and urine concentration. PMID:17522053

Zhang, Wei; Zitron, Edgar; Hömme, Meike; Kihm, Lars; Morath, Christian; Scherer, Daniel; Hegge, Stephan; Thomas, Dierk; Schmitt, Claus P; Zeier, Martin; Katus, Hugo; Karle, Christoph; Schwenger, Vedat

2007-07-20

62

Engineering Proteins for Custom Inhibition of CaV Channels  

NSDL National Science Digital Library

The influx of Ca2+ ions through voltage-dependent calcium (CaV) channels links electrical signals to physiological responses in all excitable cells. Not surprisingly, blocking CaV channel activity is a powerful method to regulate the function of excitable cells, and this is exploited for both physiological and therapeutic benefit. Nevertheless, the full potential for CaV channel inhibition is not being realized by currently available small-molecule blockers or second-messenger modulators due to limitations in targeting them either to defined groups of cells in an organism or to distinct subcellular regions within a single cell. Here, we review early efforts to engineer protein molecule blockers of CaV channels to fill this crucial niche. This technology would greatly expand the toolbox available to physiologists studying the biology of excitable cells at the cellular and systems level.

Xianghua Xu (College of Physicians and Surgeons, Columbia University Physiology and Cellular Biophysics); Henry Colecraft (Columbia University)

2009-08-01

63

Heterologous expression of tulip petal plasma membrane aquaporins in Pichia pastoris for water channel analysis.  

PubMed

Water channels formed by aquaporins (AQPs) play an important role in the control of water homeostasis in individual cells and in multicellular organisms. Plasma membrane intrinsic proteins (PIPs) constitute a subclass of plant AQPs. TgPIP2;1 and TgPIP2;2 from tulip petals are members of the PIP family. In this study, we overexpressed TgPIP2;1 and TgPIP2;2 in Pichia pastoris and monitored their water channel activity (WCA) either by an in vivo spheroplast-bursting assay performed after hypo-osmotic shock or by growth assay. Osmolarity, pH, and inhibitors of AQPs, protein kinases (PKs), and protein phosphatases (PPs) affect the WCA of heterologous AQPs in this expression system. The WCA of TgPIP2;2-expressing spheroplasts was affected by inhibitors of PKs and PPs, which indicates that the water channel of this homologue is regulated by phosphorylation in P. pastoris. From the results reported herein, we suggest that P. pastoris can be employed as a heterologous expression system to assay the WCA of PIPs and to monitor the AQP-mediated channel gating mechanism, and it can be developed to screen inhibitors/effectors of PIPs. PMID:19251885

Azad, Abul Kalam; Sawa, Yoshihiro; Ishikawa, Takahiro; Shibata, Hitoshi

2009-05-01

64

Regulation of the epithelial sodium channel by accessory proteins.  

PubMed Central

The epithelial sodium channel (ENaC) is of fundamental importance in the control of sodium fluxes in epithelial cells. Modulation of sodium reabsorption through the distal nephron ENaC is an important component in the overall control of sodium balance, blood volume and thereby of blood pressure. This is clearly demonstrated by rare genetic disorders of sodium-channel activity (Liddle's syndrome and pseudohypoaldosteronism type 1), associated with contrasting effects on blood pressure. The mineralocorticoid aldosterone is a well-established modulator of sodium-channel activity. Considerable insight has now been gained into the intracellular signalling pathways linking aldosterone-mediated changes in gene transcription with changes in ion transport. Activating pathways include aldosterone-induced proteins and especially the serum- and glucocorticoid-inducible kinase (SGK) and the small G-protein, K-Ras 2A. Targeting of the ENaC for endocytosis and degradation is now emerging as a major mechanism for the down-regulation of channel activity. Several proteins acting in concert are an intrinsic part of this process but Nedd4 (neural precursor cell expressed developmentally down-regulated 4) is of central importance. Other mechanisms known to interact with ENaC and affect sodium transport include channel-activating protease 1 (CAP-1), a membrane-anchored protein, and the cystic fibrosis transmembrane regulator. The implications of research on accessory factors controlling ENaC activity are wide-ranging. Understanding cellular mechanisms controlling ENaC activity may provide a more detailed insight not only of ion-channel abnormalities in cystic fibrosis but also of the link between abnormal renal sodium transport and essential hypertension.

Gormley, Kelly; Dong, Yanbin; Sagnella, Giuseppe A

2003-01-01

65

Explosive boiling of water in parallel micro-channels  

Microsoft Academic Search

The objective of this study is to visualize the flow pattern and to measure heat transfer coefficient during explosive boiling of water in parallel triangular micro-channels. Tests were performed in the range of inlet Reynolds number 25–60, mass flux 95–340 kg\\/m2s, and heat flux 80–330 kW\\/m2.The flow visualization showed that the behavior of long vapor bubbles, occurring in a micro-channel

G. Hetsroni; A. Mosyak; E. Pogrebnyak; Z. Segal

2005-01-01

66

Erosive dynamics of channels incised by subsurface water flow  

Microsoft Academic Search

We propose a dynamical model for channels incised into an erodible bed by subsurface water flow. The model is validated by the time-resolved topographic measurements of channel growth in a laboratory-scale experiment. Surface heights in the experiment are measured via a novel laser-aided imaging technique. The erosion rate in the model is composed of diffusive and advective components as well

Alexander E. Lobkovsky; Braunen E. Smith; Arshad Kudrolli; David C. Mohrig; Daniel H. Rothman

2007-01-01

67

Identification of chloride intracellular channel proteins in spermatozoa.  

PubMed

We have identified for the first time the presence of chloride intracellular channel (CLIC) proteins in bovine epididymal spermatozoa. CLIC1 was discovered during microsequencing of proteins that co-purified with protein phosphatase 1, PP1gamma2, in sperm extracts. In addition to CLIC1, Western blot showed that two additional CLIC family members, CLIC4 and CLIC5, are also present in spermatozoa. CLIC fusion proteins, GST-CLIC1, GST-CLIC4 and GST-CLIC5, were all able to bind to PP1gamma2 in sperm extracts during pull-down assays. Immunofluorescence microscopy revealed that each of the three isoforms occupies a distinct location within the cell. Given that PP1gamma2 is a key enzyme regulating sperm motility, PP1gamma2-binding proteins, such as the CLIC proteins, are likely to play significant roles in sperm function. PMID:15147883

Myers, Kimberley; Somanath, Payaningal R; Berryman, Mark; Vijayaraghavan, Srinivasan

2004-05-21

68

A Stochastic Model of Ionic Permeation in Protein Channels, I  

NASA Astrophysics Data System (ADS)

"A Stochastic Model of Ionic Permeation in Protein Channels, I" Z. Schuss, B. Nadler, A. Singer, R.S. Eisenberg Abstract The motion of ions through solution and protein channels is diffusion in an electrostatic field. The phenomenology of permeation through narrow channels is strongly nonlinear, exhibiting blocking, saturation, anomalous flux ratios, selectivity of different ionic species, nonlinear conductivity, and more. The confined geometry of the channels enhances the effect of ion-ion interactions in channel. This interaction is believed to be the cause of many of the nonlinear permeation phenomena. The steady-state diffusion of particles with short- and long-range interactions leads to fundamental problems in non-equilibrium statistical mechanics. In particular, continuum descriptions of such diffusion need to incorporate these interactions in order to reproduce the non-linear phenomena. The pair correlation function, analogous to that of equilibrium statistical mechanics, plays a key role in the description of ionic permeation by continuum equations. It is believed that it accounts for many of the non-linear channel phenomena, including selectivity. We show that the non-equilibrium pair correlation function of interacting ions can be determined from Poisson-Nernst-Planck equations (PNPEs) and we determine the boundary conditions it satisfies. This is a new result in non-equilibrium statistical mechanics. To decouple the PNPEs for the pair correlation function from these for the triplet correlation function, a closure relation is needed. We derive a closure relation for short-range interactions, such as Lennard-Jones repulsion, and propose an approach to the closure problem for long-range interactions, such as coulomb forces.

Schuss, Zeev

2003-03-01

69

Stability Analysis of a Uniformly Heated Channel with Supercritical Water  

SciTech Connect

The thermal-hydraulic stability of a uniformly heated channel at supercritical water pressure has been investigated to help understand the system instability phenomena which may occur in Supercritical Water Nuclear Reactors (SCWR). We have extended the modeling approach often used for Boiling Water Nuclear Reactor (BWR) stability analysis to supercritical pressure operation conditions. We have shown that Ledinegg excursive instabilities and pressure-drop oscillations (PDO) will not occur in supercritical water systems. The linear stability characteristics of a typical uniformly heated channel were computed by evaluating the eigenvalues of the model. An analysis of non-linear instability phenomena was also performed in the time domain and the dynamic bifurcations were evaluated. (authors)

Ortega Gomez, T.; Class, A.; Schulenberg, T. [Forschungszentrum Karlsruhe, Hermann-von-Helmholtz-Platz 1 76344 Eggenstein-Leopoldshafen (Germany); Lahey, R.T. Jr. [Center for Multiphase Research, Rensselaer Polytechnic, Troy, NY (United States)

2006-07-01

70

Probing Protein Channel Dynamics At The Single Molecule Level.  

NASA Astrophysics Data System (ADS)

It would be difficult to overstate the importance played by protein ion channels in cellular function. These macromolecular pores allow the passage of ions across the cellular membrane and play indispensable roles in all aspects of neurophysiology. While the patch-clamp technique continues to provide elegant descriptions of the kinetic processes involved in ion channel gating, the associated conformational changes remain a mystery. We are using the spectroscopic capabilities and single molecule fluorescence sensitivity of near-field scanning optical microscopy (NSOM) to probe these dynamics at the single channel level. Using a newly developed cantilevered NSOM probe capable of probing soft biological samples with single molecule fluorescence sensitivity, we have begun mapping the location of single NMDA receptors in intact rat cortical neurons with <100 nm spatial resolution. We will also present recent results exploring the conformational changes accompanying activation of nuclear pore channels located in the nuclear membrane of Xenopus oocytes. Our recent NSOM and AFM measurements on single nuclear pore complexes reveal large conformational changes taking place upon activation, providing rich, new molecular level details of channel function.

Lee, M. Ann; Dunn, Robert C.

1997-03-01

71

Bimodal modulation of the botulinum neurotoxin protein-conducting channel  

PubMed Central

Clostridium botulinum neurotoxin (BoNT) is the causative agent of botulism, a neuroparalytic disease. We describe here a semisynthetic strategy to identify inhibitors based on toosendanin, a traditional Chinese medicine reported to protect from BoNT intoxication. Using a single molecule assay of BoNT serotypes A and E light chain (LC) translocation through the heavy chain (HC) channel in neurons, we discovered that toosendanin and its tetrahydrofuran analog selectively arrest the LC translocation step of intoxication with subnanomolar potency, and increase the unoccluded HC channel propensity to open with micromolar efficacy. The inhibitory profile on LC translocation is accurately recapitulated in 2 different BoNT intoxication assays, namely the mouse protection and the primary rat spinal cord cell assays. Toosendanin has an unprecedented dual mode of action on the protein-conducting channel acting as a cargo-dependent inhibitor of translocation and as cargo-free channel activator. These results imply that the bimodal modulation by toosendanin depends on the dynamic interactions between channel and cargo, highlighting their tight interplay during the progression of LC transit across endosomes.

Fischer, Audrey; Nakai, Yuya; Eubanks, Lisa M.; Clancy, Colin M.; Tepp, William H.; Pellett, Sabine; Dickerson, Tobin J.; Johnson, Eric A.; Janda, Kim D.; Montal, Mauricio

2009-01-01

72

Zn2+ Interaction with Alzheimer Amyloid beta Protein Calcium Channels  

Microsoft Academic Search

The Alzheimer disease 40-residue amyloid beta protein (Abeta P[1-40]) forms cation-selective channels across acidic phospholipid bilayer membranes with spontaneous transitions over a wide range of conductances ranging from 40 to 4000 pS. Zn2+ has been reported to bind to Abeta P[1-40] with high affinity, and it has been implicated in the formation of amyloid plaques. We now report the functional

Nelson Arispe; Harvey B. Pollard; Eduardo Rojas

1996-01-01

73

ThermoTRP channels as modular proteins with allosteric gating.  

PubMed

Ion channels activate by sensing stimuli such as membrane voltage, ligand binding or temperature and transduce this information into conformational changes that open the channel pore. Thus, a key question in understanding ion channel function is how do the protein domains involved in sensing stimuli (sensors) and opening the pore (gates) communicate. In this regard, transient receptor potential (TRP) channels that confer thermosensation [A. Dhaka, V. Viswanath, A. Patapoutian, TRP ion channels and temperature sensation, Annu. Rev. Neurosci. 29 (2006) 135-161; I.S. Ramsey, M. Delling, D.E. Clapham, An introduction to TRP channels, Annu. Rev. Physiol. 68 (2006) 619-647] (thermoTRP; Q(10)>10) are unique to the extent that they integrate a variety of physical and chemical stimuli. In some cases such as, for example, the vanilloid receptor TRPV1 [M.J. Caterina, M.A. Schumacher, M. Tominaga, T.A. Rosen, J.D. Levine, D. Julius, The capsaicin receptor: a heat-activated ion channel in the pain pathway, Nature 389 (1997) 816-824] and TRPA1 [G.M. Story, A.M. Peier, A.J. Reeve, S.R. Eid, J. Mosbacher, T.R. Hricik, T.J. Earley, A.C. Hergarden, D.A. Andersson, S.W. Hwang, P. McIntyre, T. Jegla, S. Bevan, A. Patapoutian, ANKTM1, a TRP-like channel expressed in nociceptive neurons, is activated by cold temperatures, Cell 112 (2003) 819-829; S. Jordt, D. Julius, Molecular basis for species-specific sensitivity to "hot" chilli peppers, Cell 108 (2002) 421-430] the integration of these stimuli elicit pain [M. Tominaga, M.J. Caterina, A.B. Malmberg, T.A. Rosen, H. Gilbert, K. Skinner, B.E. Raumann, A.I. Basbaum, D. Julius, The cloned capsaicin receptor integrates multiple pain-producing stimuli, Neuron 21 (1998) 531-543; M. Bandell, A. Dubin, M. Petrus, A. Orth, J. Mathur, S. Hwang, A. Patapoutian, High-throughput random mutagenesis screen reveals TRPM8 residues specifically required for activation by menthol, Nat. Neurosci. 9 (2006) 466-468; S. Zurborg, B. Yurgionas, JA. Jira, O. Caspani, P.A. Heppenstall, Direct activation of the ion channel TRPA1 by Ca(2+), Nat. Neurosci. 10 (2007) 277-279]. These stimuli include voltage, pH, agonist binding, and temperature. Understanding how each of these distinct physiological signals regulate channel opening will be informative about the mechanical linkages that can act either independently or in concert to influence channel activation. In this paper we show that thermoTRP channel-forming proteins are modular in the sense that certain structure or structures (modules) confer temperature-dependent regulation, whereas others confer voltage-dependent regulation. We also discuss the thermodynamic basis of heat and cold activation in an effort to elucidate what confer to these channels the capability to be gated by temperature directly. PMID:17499848

Latorre, Ramon; Brauchi, Sebastian; Orta, Gerardo; Zaelzer, Cristián; Vargas, Guillermo

2007-01-01

74

Channelopathies from mutations in the cardiac sodium channel protein complex.  

PubMed

The cardiac sodium current underlies excitability in heart, and inherited abnormalities of the proteins regulating and conducting this current cause inherited arrhythmia syndromes. This review focuses on inherited mutations in non-pore forming proteins of sodium channel complexes that cause cardiac arrhythmia, and the deduced mechanisms by which they affect function and dysfunction of the cardiac sodium current. Defining the structure and function of these complexes and how they are regulated will contribute to understanding the possible roles for this complex in normal and abnormal physiology and homeostasis. This article is part of a Special Issue entitled "Na(+) Regulation in Cardiac Myocytes". PMID:23557754

Adsit, Graham S; Vaidyanathan, Ravi; Galler, Carla M; Kyle, John W; Makielski, Jonathan C

2013-08-01

75

Secretins: dynamic channels for protein transport across membranes  

PubMed Central

Secretins form mega-Dalton bacterial membrane channels in at least four sophisticated multi-protein systems that are crucial for translocation of proteins and assembled fibers across the outer membrane of many species of bacteria. Secretin subunits contain multiple domains, which interact with numerous other proteins, including pilotins, secretion system partner proteins and exoproteins. Our understanding of the structure of secretins is rapidly progressing, and we now recognize that features common to all secretins include a cylindrical arrangement of 12–15 subunits, a large periplasmic vestibule with a wide opening on one end and a periplasmic gate at the other end. Secretins might also play a key role in the biogenesis of their cognate secretion systems.

Korotkov, Konstantin V.; Gonen, Tamir; Hol, Wim G. J.

2011-01-01

76

Novel geminate recombination channel after indirect photoionization of water  

SciTech Connect

We studied the photolysis of neat protonated and heavy water using pump-probe and pump-repump-probe spectroscopy. A novel recombination channel is reported leading to ultrafast quenching (0.7 {+-} 0.1 ps) of almost one third of the initial number of photo-generated electrons. The efficiency and the recombination rate of this channel are lower in heavy water, 27 {+-} 5% and (0.9 {+-} 0.1 ps){sup -1}, respectively. Comparison with similar data measured after photodetachment of aqueous hydroxide provides evidence for the formation of short-lived OH:e{sup -} (OD:e{sup -}) pairs after indirect photoionization of water at 9.2 eV.

Fischer, Martin K.; Rossmadl, Hubert; Iglev, Hristo [Physik-Department E 11, Technische Universitaet Muenchen, D-85748 Garching (Germany)

2011-06-07

77

Challenging accepted ion channel biology: p64 and the CLIC family of putative intracellular anion channel proteins (Review).  

PubMed

Parchorin, p64 and the related chloride intracellular channel (CLIC) proteins are widely expressed in multicellular organisms and have emerged as candidates for novel, auto-inserting, self-assembling intracellular anion channels involved in a wide variety of fundamental cellular events including regulated secretion, cell division and apoptosis. Although the mammalian phosphoproteins p64 and parchorin (49 and 65K, respectively) have only been indirectly implicated in anion channel activity, two CLIC proteins (CLIC1 and CLIC4, 27 and 29K, respectively) appear to be essential molecular components of anion channels, and CLIC1 can form anion channels in planar lipid bilayers in the absence of other cellular proteins. However, these putative ion channel proteins are controversial because they exist in both soluble and membrane forms, with at least one transmembrane domain. Even more surprisingly, soluble CLICs share the same glutaredoxin fold as soluble omega class glutathione-S-transferases. Working out how these ubiquitous, soluble proteins unfold, insert into membranes and then refold to form integral membrane proteins, and how cells control this potentially dangerous process and make use of the associated ion channels, are challenging prospects. Critical to this future work is the need for better characterization of membrane topology, careful functional analysis of reconstituted and native channels, including their conductances and selectivities, and detailed structure/function studies including targeted mutagenesis to investigate the structure of the putative pore, the role of protein phosphorylation and the role of conserved cysteine residues. PMID:12745921

Ashley, R H

2003-01-01

78

High-Density Reconstitution of Functional Water Channels into Vesicular and Planar Block Copolymer Membranes  

PubMed Central

The exquisite selectivity and unique transport properties of membrane proteins can be harnessed for a variety of engineering and biomedical applications if suitable membranes can be produced. Amphiphilic block copolymers (BCPs), developed as stable lipid analogs, form membranes that functionally incorporate membrane proteins and are ideal for such applications. While high protein density and planar membrane morphology are most desirable, BCP–membrane protein aggregates have so far been limited to low protein densities in either vesicular or bilayer morphologies. Here, we used dialysis to reproducibly form planar and vesicular BCP membranes with a high density of reconstituted aquaporin-0 (AQP0) water channels. We show that AQP0 retains its biological activity when incorporated at high density in BCP membranes, and that the morphology of the BCP–protein aggregates can be controlled by adjusting the amount of incorporated AQP0. We also show that BCPs can be used to form two-dimensional crystals of AQP0.

2012-01-01

79

Filter gate closure inhibits ion but not water transport through potassium channels  

PubMed Central

The selectivity filter of K+ channels is conserved throughout all kingdoms of life. Carbonyl groups of highly conserved amino acids point toward the lumen to act as surrogates for the water molecules of K+ hydration. Ion conductivity is abrogated if some of these carbonyl groups flip out of the lumen, which happens (i) in the process of C-type inactivation or (ii) during filter collapse in the absence of K+. Here, we show that K+ channels remain permeable to water, even after entering such an electrically silent conformation. We reconstituted fluorescently labeled and constitutively open mutants of the bacterial K+ channel KcsA into lipid vesicles that were either C-type inactivating or noninactivating. Fluorescence correlation spectroscopy allowed us to count both the number of proteoliposomes and the number of protein-containing micelles after solubilization, providing the number of reconstituted channels per proteoliposome. Quantification of the per-channel increment in proteoliposome water permeability with the aid of stopped-flow experiments yielded a unitary water permeability pf of (6.9 ± 0.6) × 10?13 cm3?s?1 for both mutants. “Collapse” of the selectivity filter upon K+ removal did not alter pf and was fully reversible, as demonstrated by current measurements through planar bilayers in a K+-containing medium to which K+-free proteoliposomes were fused. Water flow through KcsA is halved by 200 mM K+ in the aqueous solution, which indicates an effective K+ dissociation constant in that range for a singly occupied channel. This questions the widely accepted hypothesis that multiple K+ ions in the selectivity filter act to mutually destabilize binding.

Hoomann, Torben; Jahnke, Nadin; Horner, Andreas; Keller, Sandro; Pohl, Peter

2013-01-01

80

Novel Channel Enzyme Fusion Proteins Confer Arsenate Resistance*  

PubMed Central

Steady exposure to environmental arsenic has led to the evolution of vital cellular detoxification mechanisms. Under aerobic conditions, a two-step process appears most common among microorganisms involving reduction of predominant, oxidized arsenate (H2AsVO4?/HAsVO42?) to arsenite (AsIII(OH)3) by a cytosolic enzyme (ArsC; Escherichia coli type arsenate reductase) and subsequent extrusion via ArsB (E. coli type arsenite transporter)/ACR3 (yeast type arsenite transporter). Here, we describe novel fusion proteins consisting of an aquaglyceroporin-derived arsenite channel with a C-terminal arsenate reductase domain of phosphotyrosine-phosphatase origin, providing transposable, single gene-encoded arsenate resistance. The fusion occurred in actinobacteria from soil, Frankia alni, and marine environments, Salinispora tropica; Mycobacterium tuberculosis encodes an analogous ACR3-ArsC fusion. Mutations rendered the aquaglyceroporin channel more polar resulting in lower glycerol permeability and enhanced arsenite selectivity. The arsenate reductase domain couples to thioredoxin and can complement arsenate-sensitive yeast strains. A second isoform with a nonfunctional channel may use the mycothiol/mycoredoxin cofactor pool. These channel enzymes constitute prototypes of a novel concept in metabolism in which a substrate is generated and compartmentalized by the same molecule. Immediate diffusion maintains the dynamic equilibrium and prevents toxic accumulation of metabolites in an energy-saving fashion.

Wu, Binghua; Song, Jie; Beitz, Eric

2010-01-01

81

Water-Protein Interactions: The Secret of Protein Dynamics  

PubMed Central

Water-protein interactions help to maintain flexible conformation conditions which are required for multifunctional protein recognition processes. The intimate relationship between the protein surface and hydration water can be analyzed by studying experimental water properties measured in protein systems in solution. In particular, proteins in solution modify the structure and the dynamics of the bulk water at the solute-solvent interface. The ordering effects of proteins on hydration water are extended for several angstroms. In this paper we propose a method for analyzing the dynamical properties of the water molecules present in the hydration shells of proteins. The approach is based on the analysis of the effects of protein-solvent interactions on water protons NMR relaxation parameters. NMR relaxation parameters, especially the nonselective (R1NS ) and selective (R1SE ) spin-lattice relaxation rates of water protons, are useful for investigating the solvent dynamics at the macromolecule-solvent interfaces as well as the perturbation effects caused by the water-macromolecule interactions on the solvent dynamical properties. In this paper we demonstrate that Nuclear Magnetic Resonance Spectroscopy can be used to determine the dynamical contributions of proteins to the water molecules belonging to their hydration shells.

Martini, Silvia; Bonechi, Claudia; Foletti, Alberto; Rossi, Claudio

2013-01-01

82

Aquaglyceroporins: Channel proteins with a conserved core, multiple functions, and variable surfaces  

Microsoft Academic Search

Membrane channels for water and small nonionic solutes are required for osmoregulation in bacteria, plants, and animals. Aquaporin-1, the water channel of human erythrocytes, is the first channel demonstrated to conduct water, by expression in Xenopus oocytes. Phylogenetic analyses reveal the existence of two clusters of subfamilies, the aquaporins (AQPs) and glycerol facilitators (GLPs). Sequence-based structure prediction provided a model

Andreas Engel; Henning Stahlberg

2002-01-01

83

A channeled tRNA cycle during mammalian protein synthesis.  

PubMed Central

In earlier studies it was shown that the mammalian translation system is highly organized in vivo and that the intermediates in the process, aminoacyl-tRNAs, are channeled--i.e., they are directly transferred from the aminoacyl-tRNA synthetases to the elongation factor to the ribosomes without dissociating into the cellular fluid. Here, we examine whether spent tRNAs leaving the ribosome enter the fluid phase or are transferred directly to their cognate aminoacyl-tRNA synthetases to complete a channeled tRNA cycle. Using a permeabilized CHO cell system that closely mimics living cells, we find that there is no leakage of endogenous tRNA during many cycles of translation, and protein synthesis remains linear during this period, even though free aminoacyl-tRNA is known to rapidly equilibrate between the inside and outside of these cells. We also find that exogenous tRNA and periodate-oxidized tRNA have no effect on protein synthesis in this system, indicating that they do not enter the translation machinery, despite the fact that exogenous tRNA rapidly distributes throughout the cells. Furthermore, most of the cellular aminoacyl-tRNA synthetases function only with endogenous tRNAs, although a portion can use exogenous tRNA molecules. However, aminoacylation of these exogenous tRNAs is strongly inhibited by oxidized tRNA; this inhibitor has no effect on endogenous aminoacylation. On the basis of these and the earlier observations, we conclude that endogenous tRNA is never free of the protein synthetic machinery at any stage of the translation process and, consequently, that there is a channeled tRNA cycle during protein synthesis in mammalian cells.

Stapulionis, R; Deutscher, M P

1995-01-01

84

Identification and characterization of the channel-forming protein in the cell wall of Corynebacterium amycolatum.  

PubMed

The mycolic-acid layer of certain gram-positive bacteria, the mycolata, represents an additional permeability barrier for the permeation of small water-soluble solutes. Consequently, it was shown in recent years that the mycolic acid layer of individual bacteria of the group mycolata contains pores, called porins, for the passage of hydrophilic solutes. Corynebacterium amycolatum, a pathogenic Corynebacterium species, belongs to the Corynebacteriaceae family but it lacks corynomycolic acids in its cell wall. Despite the absence of corynomycolic acids the cell wall of C. amycolatum contains a cation-selective cell wall channel, which may be responsible for the limited permeability of the cell wall of C. amycolatum. Based on partial sequencing of the protein responsible for channel formation derived from C. amycolatum ATCC 49368 we were able to identify the gene coram0001_1986 within the known genome sequence of C. amycolatum SK46 that codes for the cell wall channel. The corresponding gene of C. amycolatum ATCC 49368 was cloned into the plasmid pXHis for its expression in Corynebacterium glutamicum ?porA?porH. Biophysical characterization of the purified protein (PorAcoram) suggested that coram0001_1986 is indeed the gene coding for the pore-forming protein PorAcoram in C. amycolatum ATCC 49368. The protein belongs to the DUF (Domains of Unknown Function) 3068 superfamily of proteins, mainly found in bacteria from the family Corynebacteriaceae. The nearest relative to PorAcoram within this family is an ORF which codes for PorAcres, which was also recognized in reconstitution experiments as a channel-forming protein in Corynebacterium resistens. PMID:23811360

Soltan Mohammadi, Nafiseh; Mafakheri, Samaneh; Abdali, Narges; Bárcena-Uribarri, Iván; Tauch, Andreas; Benz, Roland

2013-11-01

85

Identification of a functional water channel in cytochrome P450 enzymes  

PubMed Central

Cytochrome P450 enzymes are monooxygenases that contain a functional heme b group linked to a conserved cysteine with a thiolate bond. In the native state, the central iron atom is hexacoordinated with a covalently bound water molecule. The exclusion of solvent molecules from the active site is essential for efficient enzymatic function. Upon substrate binding, water has to be displaced from the active site to prevent electron uncoupling that results in hydrogen peroxide or water. In contrast to typical hemoproteins, the protein surface is not directly accessible from the heme of cytochromes P450. We postulate a two-state model in which a conserved arginine, stabilizing the heme propionate in all known cytochrome P450 crystal structures, changes from the initial, stable side-chain conformation to another rotamer (metastable). In this new state, a functional water channel (aqueduct) is formed from the active site to a water cluster located on the thiolate side of the heme, close to the protein surface. This water cluster communicates with the surface in the closed state and is partly replaced by the flipping arginine side chain in the open state, allowing water molecules to exit to the surface or to reaccess the active site. This two-state model suggests the presence of an exit pathway for water between the active site and the protein surface.

Oprea, Tudor I.; Hummer, Gerhard; Garcia, Angel E.

1997-01-01

86

Atrial G Protein-Activated K^+ Channel: Expression Cloning and Molecular Properties  

Microsoft Academic Search

Activity of several ion channels is controlled by heterotrimeric GTP-binding proteins (G proteins) via a membrane-delimited pathway that does not involve cytoplasmic intermediates. The best studied example is the K^+ channel activated by muscarinic agonists in the atrium, which plays a crucial role in regulating the heartbeat. To enable studies of the molecular mechanisms of activation, this channel, denoted KGA,

Nathan Dascal; Wolfgang Schreibmayer; Nancy F. Lim; Weizhen Wang; Charles Chavkin; Lisa Dimagno; Cesar Labarca; Brigitte L. Kieffer; Claire Gaveriaxu-Ruff; David Trollinger; Henry A. Lester; Norman Davidson

1993-01-01

87

The functional interaction of accessory proteins and voltage-gated sodium channels  

Microsoft Academic Search

Sodium channels are important transmembrane proteins that underlie membrane excitability, including the excitability of neurons in pain pathways. The biophysical properties and densities of sodium channels are modulated by the presence of accessory ß-subunits, with the intracellular and extracellular binding properties of the ß1- subunit being particularly important in node of Ranvier formation. Other proteins interact with sodium channels, some

Kenji Okuse; Mark D. Baker

88

An Efficient Method for Modeling Kinetic Behavior of Channel Proteins in Cardiomyocytes  

Microsoft Academic Search

Characterization of the kinetic and conformational properties of channel proteins is a crucial element in the integrative study of congenital cardiac diseases. The proteins of the ion channels of cardiomyocytes represent an important family of biological components determining the physiology of the heart. Some computational studies aiming to understand the mechanisms of the ion channels of cardiomyocytes have concentrated on

Chong Wang; Peter Beyerlein; Heike Pospisil; Antje Krause; Chris Nugent; Werner Dubitzky

2012-01-01

89

Dual regulation of G proteins and the G-protein-activated K+ channels by lithium.  

PubMed

Lithium (Li(+)) is widely used to treat bipolar disorder (BPD). Cellular targets of Li(+), such as glycogen synthase kinase 3? (GSK3?) and G proteins, have long been implicated in BPD etiology; however, recent genetic studies link BPD to other proteins, particularly ion channels. Li(+) affects neuronal excitability, but the underlying mechanisms and the relevance to putative BPD targets are unknown. We discovered a dual regulation of G protein-gated K(+) (GIRK) channels by Li(+), and identified the underlying molecular mechanisms. In hippocampal neurons, therapeutic doses of Li(+) (1-2 mM) increased GIRK basal current (Ibasal) but attenuated neurotransmitter-evoked GIRK currents (Ievoked) mediated by Gi/o-coupled G-protein-coupled receptors (GPCRs). Molecular mechanisms of these regulations were studied with heterologously expressed GIRK1/2. In excised membrane patches, Li(+) increased Ibasal but reduced GPCR-induced GIRK currents. Both regulations were membrane-delimited and G protein-dependent, requiring both G? and G?? subunits. Li(+) did not impair direct activation of GIRK channels by G??, suggesting that inhibition of Ievoked results from an action of Li(+) on G?, probably through inhibition of GTP-GDP exchange. In direct binding studies, Li(+) promoted GPCR-independent dissociation of G?i(GDP) from G?? by a Mg(2+)-independent mechanism. This previously unknown Li(+) action on G proteins explains the second effect of Li(+), the enhancement of GIRK's Ibasal. The dual effect of Li(+) on GIRK may profoundly regulate the inhibitory effects of neurotransmitters acting via GIRK channels. Our findings link between Li(+), neuronal excitability, and both cellular and genetic targets of BPD: GPCRs, G proteins, and ion channels. PMID:24639496

Farhy Tselnicker, Isabella; Tsemakhovich, Vladimir; Rishal, Ida; Kahanovitch, Uri; Dessauer, Carmen W; Dascal, Nathan

2014-04-01

90

Protein-protein interactions in intracellular Ca2+-release channel function.  

PubMed Central

Release of Ca2+ ions from intracellular stores can occur via two classes of Ca2+-release channel (CRC) protein, the inositol 1,4, 5-trisphosphate receptors (InsP3Rs) and the ryanodine receptors (RyRs). Multiple isoforms and subtypes of each CRC class display distinct but overlapping distributions within mammalian tissues. InsP3Rs and RyRs interact with a plethora of accessory proteins which modulate the activity of their intrinsic channels. Although many aspects of CRC structure and function have been reviewed in recent years, the properties of proteins with which they interact has not been comprehensively surveyed, despite extensive current research on the roles of these modulators. The aim of this article is to review the regulation of CRC activity by accessory proteins and, wherever possible, to outline the structural details of such interactions. The CRCs are large transmembrane proteins, with the bulk of their structure located cytoplasmically. Intra- and inter-complex protein-protein interactions between these cytoplasmic domains also regulate CRC function. Some accessory proteins modulate channel activity of all CRC subtypes characterized, whereas other have class- or even isoform-specific effects. Certain accessory proteins exert both direct and indirect forms of regulation on CRCs, occasionally with opposing effects. Others are themselves modulated by changes in Ca2+ concentration, thereby participating in feedback mechanisms acting on InsP3R and RyR activity. CRCs are therefore capable of integrating numerous signalling events within a cell by virtue of such protein-protein interactions. Consequently, the functional properties of InsP3Rs and RyRs within particular cells and subcellular domains are 'customized' by the accessory proteins present.

MacKrill, J J

1999-01-01

91

Aquaporins: highly regulated channels controlling plant water relations.  

PubMed

Plant growth and development are dependent on tight regulation of water movement. Water diffusion across cell membranes is facilitated by aquaporins that provide plants with the means to rapidly and reversibly modify water permeability. This is done by changing aquaporin density and activity in the membrane, including posttranslational modifications and protein interaction that act on their trafficking and gating. At the whole organ level aquaporins modify water conductance and gradients at key "gatekeeper" cell layers that impact on whole plant water flow and plant water potential. In this way they may act in concert with stomatal regulation to determine the degree of isohydry/anisohydry. Molecular, physiological, and biophysical approaches have demonstrated that variations in root and leaf hydraulic conductivity can be accounted for by aquaporins but this must be integrated with anatomical considerations. This Update integrates these data and emphasizes the central role played by aquaporins in regulating plant water relations. PMID:24449709

Chaumont, François; Tyerman, Stephen D

2014-04-01

92

The role of transmembrane channel-like proteins in the operation of hair cell mechanotransducer channels  

PubMed Central

Sound stimuli elicit movement of the stereocilia that make up the hair bundle of cochlear hair cells, putting tension on the tip links connecting the stereocilia and thereby opening mechanotransducer (MT) channels. Tmc1 and Tmc2, two members of the transmembrane channel–like family, are necessary for mechanotransduction. To assess their precise role, we recorded MT currents elicited by hair bundle deflections in mice with null mutations of Tmc1, Tmc2, or both. During the first postnatal week, we observed a normal MT current in hair cells lacking Tmc1 or Tmc2; however, in the absence of both isoforms, we recorded a large MT current that was phase-shifted 180°, being evoked by displacements of the hair bundle away from its tallest edge rather than toward it as in wild-type hair cells. The anomalous MT current in hair cells lacking Tmc1 and Tmc2 was blocked by FM1-43, dihydrostreptomycin, and extracellular Ca2+ at concentrations similar to those that blocked wild type. MT channels in the double knockouts carried Ca2+ with a lower permeability than wild-type or single mutants. The MT current in double knockouts persisted during exposure to submicromolar Ca2+, even though this treatment destroyed the tip links. We conclude that the Tmc isoforms do not themselves constitute the MT channel but are essential for targeting and interaction with the tip link. Changes in the MT conductance and Ca2+ permeability observed in the absence of Tmc1 mutants may stem from loss of interaction with protein partners in the transduction complex.

Kim, Kyunghee X.; Beurg, Maryline; Hackney, Carole M.; Furness, David N.; Mahendrasingam, Shanthini

2013-01-01

93

Aquaporin-4 water channel oligomers are associated with the transverse tubules of skeletal myofibers.  

PubMed

Transverse (T) tubules comprise a tortuous network inside the skeletal myofibers enclosing a distinct osmotic environment. Here we have examined whether the T tubules contain aquaporin type 4 (AQP4) water channels to mediate rapid transmembrane water flow. Separation of T tubular and sarcolemmal membranes by sucrose density gradient centrifugation revealed that two main isoforms of AQP4, namely M23 and M1, were present in both membrane fractions. Compatible with this, expression of fluorescent Venus-AQP4.M23 in rat muscle showed the protein both in the T tubules and at the sarcolemma. Blue-Native polyacrylamide gel electrophoresis showed that higher order oligomers typical to the AQP4 water channel were present in both membrane compartments. Interestingly, ?-syntrophin that mediates binding of AQP4 to the sarcolemmal dystrophin glycoprotein complex was also present in the T tubule fraction. Deletion of the syntrophin-binding sequence of AQP4 increased its mobile fraction at the sarcolemma but not in the T tubules. Taken together, our results strongly suggest that both the sarcolemma and the T tubules harbor higher order oligomers of the AQP4 water channel but the interactions with adjacent macromolecules are different. PMID:20849844

Kaakinen, Mika; Zelenin, Sergei; Metsikkö, Kalervo

2011-01-01

94

Actin directly interacts with different membrane channel proteins and influences channel activities: AQP2 as a model.  

PubMed

The interplay between actin and 10 membrane channel proteins that have been shown to directly bind to actin are reviewed. The 10 membrane channel proteins covered in this review are aquaporin 2 (AQP2), cystic fibrosis transmembrane conductance regulator (CFTR), ClC2, short form of ClC3 (sClC3), chloride intracellular channel 1 (CLIC1), chloride intracellular channel 5 (CLIC5), epithelial sodium channel (ENaC), large-conductance calcium-activated potassium channel (Maxi-K), transient receptor potential vanilloid 4 (TRPV4), and voltage-dependent anion channel (VDAC), with particular attention to AQP2. In regard to AQP2, most reciprocal interactions between actin and AQP2 occur during intracellular trafficking, which are largely mediated through indirect binding. Actin and the actin cytoskeleton work as cables, barriers, stabilizers, and force generators for motility. However, as with ENaC, the effects of actin cytoskeleton on channel gating should be investigated further. This article is part of a Special Issue entitled: Reciprocal influences between cell cytoskeleton and membrane channels, receptors and transporters. Guest Editor: Jean Claude Hervé. PMID:23770358

Sasaki, Sei; Yui, Naofumi; Noda, Yumi

2014-02-01

95

Existence of a tightly regulated water channel in Saccharomyces cerevisiae.  

PubMed

The Saccharomyces cerevisiae strain Sigma1278b possesses two putative aquaporins, Aqy1-1p and Aqy2-1p. Previous work demonstrated that Aqy1-1p functions as a water channel in Xenopus oocyte. However, no function could be attributed to Aqy2-1p in this system. Specific antibodies were used to follow the expression of Aqy1-1p and Aqy2-1p in the yeast. Aqy1-1p was never detected whatever the growth phase and culture conditions tested. In contrast, Aqy2-1p was detected only during the exponential growth phase in rich medium containing glucose. Aqy2-1p expression was repressed by hyper-osmotic culture conditions. Both immunocytochemistry and biochemical subcellular fractionation demonstrated that Aqy2-1p is located on the endoplasmic reticulum (ER) as well as on the plasma membrane. In microsomal vesicles enriched in ER, a water channel activity due to Aqy2-1p was detected by stopped-flow analysis. Our results show that the expression of aquaporins is tightly controlled. The physiological relevance of aquaporin-mediated water transport in yeast is discussed. PMID:11168368

Meyrial, V; Laizé, V; Gobin, R; Ripoche, P; Hohmann, S; Tacnet, F

2001-01-01

96

Molecular mechanism of H+ conduction in the single-file water chain of the gramicidin channel.  

PubMed Central

The conduction of protons in the hydrogen-bonded chain of water molecules (or "proton wire") embedded in the lumen of gramicidin A is studied with molecular dynamics free energy simulations. The process may be described as a "hop-and-turn" or Grotthuss mechanism involving the chemical exchange (hop) of hydrogen nuclei between hydrogen-bonded water molecules arranged in single file in the lumen of the pore, and the subsequent reorganization (turn) of the hydrogen-bonded network. Accordingly, the conduction cycle is modeled by two complementary steps corresponding respectively to the translocation 1) of an ionic defect (H+) and 2) of a bonding defect along the hydrogen-bonded chain of water molecules in the pore interior. The molecular mechanism and the potential of mean force are analyzed for each of these two translocation steps. It is found that the mobility of protons in gramicidin A is essentially determined by the fine structure and the dynamic fluctuations of the hydrogen-bonded network. The translocation of H+ is mediated by spontaneous (thermal) fluctuations in the relative positions of oxygen atoms in the wire. In this diffusive mechanism, a shallow free-energy well slightly favors the presence of the excess proton near the middle of the channel. In the absence of H+, the water chain adopts either one of two polarized configurations, each of which corresponds to an oriented donor-acceptor hydrogen-bond pattern along the channel axis. Interconversion between these two conformations is an activated process that occurs through the sequential and directional reorientation of water molecules of the wire. The effect of hydrogen-bonding interactions between channel and water on proton translocation is analyzed from a comparison to the results obtained previously in a study of model nonpolar channels, in which such interactions were missing. Hydrogen-bond donation from water to the backbone carbonyl oxygen atoms lining the pore interior has a dual effect: it provides a coordination of water molecules well suited both to proton hydration and to high proton mobility, and it facilitates the slower reorientation or turn step of the Grotthuss mechanism by stabilizing intermediate configurations of the hydrogen-bonded network in which water molecules are in the process of flipping between their two preferred, polarized states. This mechanism offers a detailed molecular model for the rapid transport of protons in channels, in energy-transducing membrane proteins, and in enzymes.

Pomes, Regis; Roux, Benoit

2002-01-01

97

Water channel simulation of the atmospheric boundary layer  

NASA Astrophysics Data System (ADS)

As part of a programme of work designed to assess the feasibility of modelling the dispersion of heavy plumes in a water channel it has been necessary to develop artificially structured shear layers which attempt to simulate atmospheric conditions. For a variety of reasons the choice of simulation is similar to that developed by Counihan (1969) and consists of a rough surface preceded by a castellated barrier and a number of profiled vorticity generators. Mean velocity and turbulence distributions, together with turbulent spectra and integral length scales, compared favourably with boundary layers modelled in wind tunnels and with full scale experiments in rural surroundings.

Cheah, S. C.; Cleaver, J. W.; Millward, A.

98

Selectivity and conductance among the glycerol and water conducting aquaporin family of channels.  

PubMed

The atomic structures of a transmembrane water plus glycerol conducting channel (GlpF), and now of aquaporin Z (AqpZ) from the same species, Escherichia coli, bring the total to three atomic resolution structures in the aquaporin (AQP) family. Members of the AQP family each assemble as tetramers of four channels. Common helical axes support a wider channel in the glycerol plus water channel paradigm, GlpF. Water molecules form a single hydrogen bonded file throughout the 28 A long channel in AqpZ. The basis for absolute exclusion of proton or hydronium ion conductance through the line of water is explored using simulations. PMID:14630323

Stroud, Robert M; Savage, David; Miercke, Larry J W; Lee, John K; Khademi, Shahram; Harries, William

2003-11-27

99

Highly selective water channel activity measured by voltage clamp: Analysis of planar lipid bilayers reconstituted with purified AqpZ  

PubMed Central

Aquaporins are membrane channels selectively permeated by water or water plus glycerol. Conflicting reports have described ion conductance associated with some water channels, raising the question of whether ion conductance is a general property of the aquaporin family. To clarify this question, a defined system was developed to simultaneously measure water permeability and ion conductance. The Escherichia coli water channel aquaporin-Z (AqpZ) was studied, because it is a highly stable tetramer. Planar lipid bilayers were formed from unilamellar vesicles containing purified AqpZ. The hydraulic conductivity of bilayers made from the total extract of E. coli lipids increased 3-fold if reconstituted with AqpZ, but electric conductance was unchanged. No channel activity was detected under voltage-clamp conditions, indicating that less than one in 109 transport events is electrogenic. Microelectrode measurements were simultaneously undertaken adjacent to the membrane. Changes in sodium concentration profiles accompanying transmembrane water flow permitted calculation of the activation energies: 14 kcal/mol for protein-free lipid bilayers and 4 kcal/mol for lipid bilayers containing AqpZ. Neither the water permeability nor the electric conductivity exhibited voltage dependence. This sensitive system demonstrated that AqpZ is permeated by water but not charged ions and should permit direct analyses of putative electrogenic properties of other aquaporins.

Pohl, Peter; Saparov, Sapar M.; Borgnia, Mario J.; Agre, Peter

2001-01-01

100

Ligand-gated ion channel interacting proteins and their role in neuroprotection  

PubMed Central

Ion channel receptors are a vital component of nervous system signaling, allowing rapid and direct conversion of a chemical neurotransmitter message to an electrical current. In recent decades, it has become apparent that ionotropic receptors are regulated by protein-protein interactions with other ion channels, G-protein coupled receptors and intracellular proteins. These other proteins can also be modulated by these interactions with ion channel receptors. This bidirectional functional cross-talk is important for critical cellular functions such as excitotoxicity in pathological and disease states like stroke, and for the basic dynamics of activity-dependent synaptic plasticity. Protein interactions with ion channel receptors can therefore increase the computational capacity of neuronal signaling cascades and also represent a novel target for therapeutic intervention in neuropsychiatric disease. This review will highlight some examples of ion channel receptor interactions and their potential clinical utility for neuroprotection.

Li, Shupeng; Wong, Albert H. C.; Liu, Fang

2014-01-01

101

Alcohol modulation of G-protein-gated inwardly rectifying potassium channels: from binding to therapeutics  

PubMed Central

Alcohol (ethanol)-induced behaviors may arise from direct interaction of alcohol with discrete protein cavities within brain proteins. Recent structural and biochemical studies have provided new insights into the mechanism of alcohol-dependent activation of G protein-gated inwardly rectifying potassium (GIRK) channels, which regulate neuronal responses in the brain reward circuit. GIRK channels contain an alcohol binding pocket formed at the interface of two adjacent channel subunits. Here, we discuss the physiochemical properties of the alcohol pocket and the roles of G protein ?? subunits and membrane phospholipid PIP2 in regulating the alcohol response of GIRK channels. Some of the features of alcohol modulation of GIRK channels may be common to other alcohol-sensitive brain proteins. We discuss the possibility of alcohol-selective therapeutics that block alcohol access to the pocket. Understanding alcohol recognition and modulation of brain proteins is essential for development of therapeutics for alcohol abuse and addiction.

Bodhinathan, Karthik; Slesinger, Paul A.

2014-01-01

102

Suppression of aquaporin, a mediator of water channel control in the carcinogenic liver fluke, Opisthorchis viverrini  

PubMed Central

Background Opisthorchiasis and Opisthorchis viverrini-associated bile duct cancer represent major public health threats in Thailand and Laos. The tegument of this food borne fluke plays pivotal roles in parasite metabolism, homeostasis and osmoregulation. Excretory/secretory products also pass from the fluke to the biliary environment, products that likely underlie pathogenesis of liver fluke infection. Aquaporins (AQPs), belong to the major intrinsic protein superfamily of integral plasma membrane channel proteins that selectively transport water across cell membranes. AQPs play key roles as water and ion transport channels through the tegument of helminth parasites. Methods Here, two forms of AQP mRNAs from the adult developmental stage of O. viverrini, termed O. viverrini aquaporin-1 and -2 (Ov-aqp-1 and -2) were investigated. Roles of Ov-aqp-1 and -2 in the movement of water across the tegument of this carcinogenic liver fluke were investigated using RNA interference. Results Ov-AQP-1 and Ov-AQP-2 contain unique characteristic asparagine-proline-alanine (NPA) motifs of AQP transmembrane proteins. Phylogenetic analysis indicated that Ov-AQPs belong to an expanding group of aquaglyceroporin-like water channel proteins characterized from helminth and protozoan parasites, which is pivotal to the specialized requirements of water and solute control during parasitism. Elevated transcription of Ov-aqp-1 was evident in the egg, cercaria, metacercaria and adult stages of O. viverrini, whereas Ov-aqp-2 transcripts were detected at higher level in egg, metacercaria, cercaria and adult stage, respectively. RNA interference using electroporated dsRNA suppressed transcript levels of Ov-aqp-1 and Ov-aqp-2 in adult worms by 58-99% over periods of up to 16 days in vitro. Suppression of Ov-aqp-1 and Ov-aqp-2 in vitro disabled water transport in adult flukes. Conclusion The apparently pivotal roles of Ov-AQP in solute homeostasis at the fluke surface suggest that deeper investigation will be informative for the pathophysiology of O. viverrini, and may uncover intervention targets, particularly in view of the singularly notable predilection of this pathogen for residence within ducts of the biliary tree.

2014-01-01

103

Time-correlation analysis of simulated water motion in flexible and rigid gramicidin channels.  

PubMed

Molecular dynamics simulations have been done on a system consisting of the polypeptide membrane channel former gramicidin, plus water molecules in the channel and caps of waters at the two ends of the channel. In the absence of explicit simulation of the surrounding membrane, the helical form of the channel was maintained by artificial restraints on the peptide motion. The characteristic time constant of the artificial restraint was varied to assess the effect of the restraints on the channel structure and water motions. Time-correlation analysis was done on the motions of individual channel waters and on the motions of the center of mass of the channel waters. It is found that individual water molecules confined in the channel execute higher frequency motions than bulk water, for all degrees of channel peptide restraint. The center-of-mass motion of the chain of channel waters (which is the motion that is critical for transmembrane transport, due to the mandatory single filing of water in the channel) does not exhibit these higher frequency motions. The mobility of the water chain is dramatically reduced by holding the channel rigid. Thus permeation through the channel is not like flow through a rigid pipe; rather permeation is facilitated by peptide motion. For the looser restraints we used, the mobility of the water chain was not very much affected by the degree of restraint. Depending on which set of experiments is considered, the computed mobility of our water chain in the flexible channel is four to twenty times too high to account for the experimentally measured resistance of the gramicidin channel to water flow. From this result it appears likely that the peptide motions of an actual gramicidin channel embedded in a lipid membrane may be more restrained than in our flexible channel model, and that these restraints may be a significant modulator of channel permeability. For the completely rigid channel model the "trapping" of the water molecules in preferred positions throughout the molecular dynamics run precludes a reasonable assessment of mobility, but it seems to be quite low. PMID:1715766

Chiu, S W; Jakobsson, E; Subramaniam, S; McCammon, J A

1991-07-01

104

Leucine zipper domain targets cAMP-dependent protein kinase to mammalian BK channels.  

PubMed

Large conductance, calcium- and voltage-activated potassium (BK) channels control excitability in many tissues and are regulated by several protein kinases and phosphatases that remain associated with the channels in cell-free patches of membrane. Here, we report the identification of a highly conserved, non-canonical, leucine zipper (LZ1) in the C terminus of mammalian BK channels that is required for cAMP-dependent protein kinase (PKA) to associate with the channel and regulate its activity. A synthetic polypeptide encompassing the central d position leucine residues in LZ1 blocks the regulation of recombinant mouse BK channels by endogenous PKA in HEK293 cells. In contrast, neither an alanine-substituted LZ1 peptide nor a peptide corresponding to another, more C-terminal putative leucine zipper, LZ2, had any effect on regulation of the channels by endogenous PKA. Mutagenesis of the central two LZ1 d position leucines to alanine in the BK channel also eliminated regulation by endogenous PKA in HEK293 cells without altering the channel sensitivity to activation by voltage or by exogenous purified PKA. Inclusion of the STREX splice insert in the BK channel protein, which switches channel regulation by PKA from stimulation to inhibition, did not alter the requirement for an intact LZ1. Although PKA does not bind directly to the channel protein in vitro, mutation of LZ1 abolished co-immunoprecipitation of PKA and the respective BK channel splice variant from HEK293 cells. Furthermore, a 127-amino acid fusion protein encompassing the functional LZ1 domain co-immunoprecipitates a PKA-signaling complex from rat brain. Thus LZ1 is required for the association and regulation of mammalian BK channels by PKA, and other putative leucine zippers in the BK channel protein may provide anchoring for other regulatory enzyme complexes. PMID:12509433

Tian, Lijun; Coghill, Lorraine S; MacDonald, Stephen H-F; Armstrong, David L; Shipston, Michael J

2003-03-01

105

Eukaryotic integral membrane protein expression utilizing the Escherichia coli glycerol-conducting channel protein (GlpF)  

Microsoft Academic Search

A fusion protein expression system is described that allows for production of eukaryotic integral membrane proteins in Escherichia coli (E. coli). The eukaryotic membrane protein targets are fused to the C terminus of the highly expressed E. coli inner membrane protein, GlpF (the glycerol-conducting channel protein). The generic utility of this system for heterologous\\u000a membrane-protein expression is demonstrated by the

Irene Neophytou; Richard Harvey; Jayne Lawrence; Phil Marsh; Barry Panaretou; David Barlow

2007-01-01

106

COALBED METHANE DISCHARGE WATER INTERACTION WITH STREAM CHANNEL SEDIMENT IN THE POWDER RIVER BASIN  

Microsoft Academic Search

Extraction of methane (natural gas) from coal deposits is facilitated by pumping of aquifer water. Coalbed methane (CBM) product water, produced from pumping groundwater, is discharged into associated unlined holding ponds or downstream channels. The objective of this study was to examine the chemistry of CBM discharge water reacting with an ephemeral stream channel sediment in the Powder River Basin,

Marji Pat; Katta J. Reddy; Quentin D. Skinner

107

Biological gas channels for NH3 and CO2: evidence that Rh (Rhesus) proteins are CO2 channels.  

PubMed

Physiological evidence from our laboratory indicates that Amt/Mep proteins are gas channels for NH3, the first biological gas channels to be described. This view has now been confirmed by structural evidence and is displacing the previous belief that Amt/Mep proteins were active transporters for the NH4+ ion. Still disputed is the physiological substrate for Rh proteins, the only known homologues of Amt/Mep proteins. Many think they are mammalian ammonium (NH4+ or NH3) transporters. Following Monod's famous dictum, "Anything found to be true of E. coli must also be true of elephants" [Perspect. Biol. Med. 47(1) (2004) 47], we explored the substrate for Rh proteins in the unicellular green alga Chlamydomonas reinhardtii. C. reinhardtii is one of the simplest organisms to have Rh proteins and it also has Amt proteins. Physiological studies in this microbe indicate that the substrate for Rh proteins is CO2 and confirm that the substrate for Amt proteins is NH3. Both are readily hydrated gases. Knowing that transport of CO2 is the ancestral function of Rh proteins supports the inference from hematological research that a newly evolving role of the human Rh30 proteins, RhCcEe and RhD, is to help maintain the flexible, flattened shape of the red cell. PMID:16563833

Kustu, S; Inwood, W

2006-01-01

108

Immunohistochemical Localization of the Water Channels AQP4 and AQP5 in the Rat Pituitary Gland  

PubMed Central

The pituitary gland is composed of the adenohypophysis and neurohypophysis. The adenohypophysis contains endocrine cells, folliculo-stellate (FS) cells, and marginal layer cells, whereas the neurohypophysis mainly comprises axons and pituicytes. To understand the molecular nature of water transfer in the pituitary gland, we examined the immunohistochemical localization of the membrane water channels aquaporin-4 (AQP4) and AQP5 in rat tissue. Double immunofluorescence analysis of AQP4 and S100 protein, a known marker for FS cells, marginal layer cells, and pituicytes, clearly revealed that FS cells and marginal layer cells in the adenohypophysis and the pituicytes in pars nervosa are positive for AQP4. AQP5 was found to be localized at the apical membrane in some marginal layer cells surrounding the Rathke’s residual pouch, in which AQP4 was observed to be localized on the basolateral membranes. These results suggest the following possibilities: 1) FS cells especially require water for their functions and 2) transepithelial water transfer could occur between the lumen of Rathke’s residual pouch and the interstitial fluid in the adenohypophysis through the AQP4 and AQP5 channels in the marginal layer cells.

Matsuzaki, Toshiyuki; Inahata, Yuki; Sawai, Nobuhiko; Yang, Chun-Ying; Kobayashi, Makito; Takata, Kuniaki; Ozawa, Hitoshi

2011-01-01

109

Immunohistochemical Localization of the Water Channels AQP4 and AQP5 in the Rat Pituitary Gland.  

PubMed

The pituitary gland is composed of the adenohypophysis and neurohypophysis. The adenohypophysis contains endocrine cells, folliculo-stellate (FS) cells, and marginal layer cells, whereas the neurohypophysis mainly comprises axons and pituicytes. To understand the molecular nature of water transfer in the pituitary gland, we examined the immunohistochemical localization of the membrane water channels aquaporin-4 (AQP4) and AQP5 in rat tissue. Double immunofluorescence analysis of AQP4 and S100 protein, a known marker for FS cells, marginal layer cells, and pituicytes, clearly revealed that FS cells and marginal layer cells in the adenohypophysis and the pituicytes in pars nervosa are positive for AQP4. AQP5 was found to be localized at the apical membrane in some marginal layer cells surrounding the Rathke's residual pouch, in which AQP4 was observed to be localized on the basolateral membranes. These results suggest the following possibilities: 1) FS cells especially require water for their functions and 2) transepithelial water transfer could occur between the lumen of Rathke's residual pouch and the interstitial fluid in the adenohypophysis through the AQP4 and AQP5 channels in the marginal layer cells. PMID:22282586

Matsuzaki, Toshiyuki; Inahata, Yuki; Sawai, Nobuhiko; Yang, Chun-Ying; Kobayashi, Makito; Takata, Kuniaki; Ozawa, Hitoshi

2011-12-28

110

Concentrating Toxoplasma gondii and Cyclospora cayetanensis from Surface Water and Drinking Water by Continuous Separation Channel Centrifugation  

EPA Science Inventory

Aims: To evaluate the effectiveness of continuous separation channel centrifugation for concentrating Toxoplasma gondii and Cyclospora cayetanensis from drinking water and environmental waters. Methods and Results: Ready-to-seed vials with known quantities of Toxoplasma gondii a...

111

Selectivity and conductance among the glycerol and water conducting aquaporin family of channels  

Microsoft Academic Search

The atomic structures of a transmembrane water plus glycerol conducting channel (GlpF), and now of aquaporin Z (AqpZ) from the same species, Escherichia coli, bring the total to three atomic resolution structures in the aquaporin (AQP) family. Members of the AQP family each assemble as tetramers of four channels. Common helical axes support a wider channel in the glycerol plus

Robert M. Stroud; David Savage; Larry J. W. Miercke; John K. Lee; Shahram Khademi; William Harries

2003-01-01

112

A novel approach for assessing protein synthesis in channel catfish, Ictalurus punctatus.  

PubMed

A comprehensive understanding of animal growth requires adequate knowledge of protein synthesis (PS), which in fish, has traditionally been determined by the flooding dose method. However, this procedure is limited to short-term assessments and may not accurately describe fish growth over extended periods of time. Since deuterium oxide ((2)H(2)O) has been used to non-invasively quantify PS in mammals over short- and long-term periods, we aimed at determining if (2)H(2)O could also be used to measure PS in channel catfish. Fish were stocked in a 40-L aquarium with approximately 4% (2)H(2)O and sampled at 4, 8 and 24h (n=6 at each time period) to determine (2)H-labeling of body water (plasma), as well as protein-free and protein-bound (2)H-labeled alanine. The labeling of body water reflected that of aquarium water and the labeling of protein-free alanine remained constant over 24h and was approximately 3.8 times greater than that of body water. By measuring (2)H-labeled alanine incorporation after 24h of (2)H(2)O exposure we were able to calculate a rate of PS: 0.04+/-0.01% h(-1). These results demonstrate that PS in fish can be effectively measured using (2)H(2)O and, because this method yields integrative measures of PS, is relatively inexpensive and accounts for perturbations such as feeding, it is a novel and practical assessment option. PMID:19563906

Gasier, Heath G; Previs, Stephen F; Pohlenz, Camilo; Fluckey, James D; Gatlin, Delbert M; Buentello, J Alejandro

2009-10-01

113

Structural basis for allosteric coupling at the membrane-protein interface in Gloeobacter violaceus ligand-gated ion channel (GLIC).  

PubMed

Ligand binding at the extracellular domain of pentameric ligand-gated ion channels initiates a relay of conformational changes that culminates at the gate within the transmembrane domain. The interface between the two domains is a key structural entity that governs gating. Molecular events in signal transduction at the interface are poorly defined because of its intrinsically dynamic nature combined with functional modulation by membrane lipid and water vestibules. Here we used electron paramagnetic resonance spectroscopy to delineate protein motions underlying Gloeobacter violaceus ligand-gated ion channel gating in a membrane environment and report the interface conformation in the closed and the desensitized states. Extensive intrasubunit interactions were observed in the closed state that are weakened upon desensitization and replaced by newer intersubunit contacts. Gating involves major rearrangements of the interfacial loops, accompanied by reorganization of the protein-lipid-water interface. These structural changes may serve as targets for modulation of gating by lipids, alcohols, and amphipathic drug molecules. PMID:24338475

Velisetty, Phanindra; Chalamalasetti, Sreevatsa V; Chakrapani, Sudha

2014-01-31

114

[Ion channels--target proteins for some animal toxins].  

PubMed

Advances in molecular biology along with improvements in electrophysiological techniques have increased the knowledge in the structure and function of many ion channels. Voltage-gated Na+ and K+ channels may become sensitive to biologically active substances such as animal toxins. Study of animal toxins can help to understand the molecular mechanisms of their action, but it can also reveal the tools for the future study of molecular physiology of ion channels. PMID:10568075

Patocka, J; Strunecká, A

1999-08-01

115

Mechanosensitivity of GIRK channels is mediated by protein kinase C-dependent channel-phosphatidylinositol 4,5-bisphosphate interaction.  

PubMed

Gprotein-activated inwardly rectifying K+ channel (GIRK or Kir3) currents are inhibited by mechanical stretch of the cell membrane, but the underlying mechanisms are not understood. In Xenopus oocytes heterologously expressing GIRK channels, membrane stretch induced by 50% reduction of osmotic pressure caused a prompt reduction of GIRK1/4, GIRK1, and GIRK4 currents by 16.6-42.6%. Comparable GIRK current reduction was produced by protein kinase C (PKC) activation (phorbol 12-myristate 13-acetate). The mechanosensitivity of the GIRK4 current was abolished by pretreatment with PKC inhibitors (staurosporine or calphostin C). Neither hypo-osmotic challenge nor PKC activation affected IRK1 currents. GIRK4 chimera (GIRK4-IRK1-(Lys207-Leu245)) and single point mutant (GIRK4(I229L)), in which the phosphatidylinositol 4,5-bisphosphate (PIP2) binding domain or residue was replaced by the corresponding region of IRK1 to strengthen the channel-PIP2 interaction, showed no mechanosensitivity and minimal PKC sensitivity. IRK1 gained mechanosensitivity and PKC sensitivity by reverse double point mutation of the PIP2 binding domain (L222I/R213Q). Overexpression of Gbetagamma, which is known to strengthen the channel-PIP2 interaction, attenuated the mechanosensitivity of GIRK4 channels. In oocytes expressing a pleckstrin homology domain of PLC-delta tagged with green fluorescent protein, hypo-osmotic challenge or PKC activation caused a translocation of the fluorescence signal from the cell membrane to the cytosol, reflecting PIP2 hydrolysis. The translocation was prevented by pretreatment with PKC inhibitors. Involvement of PKC activation in the mechanosensitivity of muscarinic K+ channels was confirmed in native rabbit atrial myocytes. These results suggest that the mechanosensitivity of GIRK channels is mediated primarily by channel-PIP2 interaction, with PKC playing an important role in modulating the interaction probably through PIP2 hydrolysis. PMID:14660621

Zhang, Liyan; Lee, Jong-Kook; John, Scott A; Uozumi, Nobuyuki; Kodama, Itsuo

2004-02-20

116

Porin channels in Escherichia coli: studies with liposomes reconstituted from purified proteins.  

PubMed Central

Rates of diffusion of uncharged and charged solute molecules through porin channels were determined by using liposomes reconstituted from egg phosphatidylcholine and purified Escherichia coli porins OmpF (protein 1a), OmpC (protein 1b), and PhoE (protein E). All three porin proteins appeared to produce channels of similar size, although the OmpF channel appeared to be 7 to 9% larger than the OmpC and PhoE channels in an equivalent radius. Hydrophobicity of the solute retarded the penetration through all three channels in a similar manner. The presence of one negative charge on the solute resulted in about a threefold reduction in penetration rates through OmpF and OmpC channels, whereas it produced two- to tenfold acceleration of diffusion through the PhoE channel. The addition of the second negatively charged group to the solutes decreased the diffusion rates through OmpF and OmpC channels further, whereas diffusion through the PhoE channel was not affected much. These results suggest that PhoE specializes in the uptake of negatively charged solutes. At the present level of resolution, no sign of true solute specificity was found in OmpF and OmpC channels; peptides, for example, diffused through both of these channels at rates expected from their molecular size, hydrophobicity, and charge. However, the OmpF porin channel allowed influx of more solute molecules per unit time than did the equivalent weight of the OmpC porin when the flux was driven by a concentration gradient of the same size. This apparent difference in "efficiency" became more pronounced with larger solutes, and it is likely to be the consequence of the difference in the sizes of OmpF and OmpC channels.

Nikaido, H; Rosenberg, E Y

1983-01-01

117

Cytoskeletal and scaffolding proteins as structural and functional determinants of TRP channels.  

PubMed

Transient receptor potential (TRP) channels are six transmembrane-spanning proteins, with variable selectivity for cations, that play a relevant role in intracellular Ca(2+) homeostasis. There is a large body of evidence that shows association of TRP channels with the actin cytoskeleton or even the microtubules and demonstrating the functional importance of this interaction for TRP channel function. Conversely, cation currents through TRP channels have also been found to modulate cytoskeleton rearrangements. The interplay between TRP channels and the cytoskeleton has been demonstrated to be essential for full activation of a variety of cellular functions. Furthermore, TRP channels have been reported to take part of macromolecular complexes including different signal transduction proteins. Scaffolding proteins play a relevant role in the association of TRP proteins with other signaling molecules into specific microdomains. Especially relevant are the roles of the Homer family members for the regulation of TRPC channel gating in mammals and INAD in the modulation of Drosophila TRP channels. This article is part of a Special Issue entitled: Reciprocal influences between cell cytoskeleton and membrane channels, receptors and transporters. Guest Editor: Jean Claude Hervé. PMID:23333715

Smani, Tarik; Dionisio, Natalia; López, José J; Berna-Erro, Alejandro; Rosado, Juan A

2014-02-01

118

Colon water transport in transgenic mice lacking aquaporin-4 water channels  

PubMed Central

Transgenic null mice were used to test the hypothesis that water channel aquaporin-4 (AQP4) is involved in colon water transport and fecal dehydration. AQP4 was immunolocalized to the basolateral membrane of colonic surface epithelium of wild-type (+/+) mice and was absent in AQP4 null (?/?) mice. The transepithelial osmotic water permeability coefficient (Pf) of in vivo perfused colon of +/+ mice, measured using the volume marker 14C-labeled polyethylene glycol, was 0.016 ± 0.002 cm/s. Pf of proximal colon was greater than that of distal colon (0.020 ± 0.004 vs. 0.009 ± 0.003 cm/s, P < 0.01). Pf was significantly lower in ?/? mice when measured in full-length colon (0.009 ± 0.002 cm/s, P < 0.05) and proximal colon (0.013 ± 0.002 cm/s, P < 0.05) but not in distal colon. There was no difference in water content of cecal stool from +/+ vs. ?/? mice (0.80 ± 0.01 vs. 0.81 ± 0.01), but there was a slightly higher water content in defecated stool from +/+ mice (0.68 ± 0.01 vs. 0.65 ± 0.01, P < 0.05). Despite the differences in water permeability with AQP4 deletion, theophylline-induced secretion was not impaired (50 ± 9 vs. 51 ± 8 ?l · min?1 · g?1). These results provide evidence that transcellular water transport through AQP4 water channels in colonic epithelium facilitates transepithelial osmotic water permeability but has little or no effect on colonic fluid secretion or fecal dehydration.

WANG, KASPER S.; MA, TONGHUI; FILIZ, FERDA; VERKMAN, A. S.; BASTIDAS, J. AUGUSTO

2012-01-01

119

EFFECT OF ELEVATED WATER TEMPERATURE ON INSECT EMERGENCE IN OUTDOOR EXPERIMENTAL CHANNELS  

EPA Science Inventory

Emergence of adult aquatic insects was evaluated weekly with plexiglass traps positioned in two outdoor experimental channels from April through August 1977. One channel was seasonal Mississippi River water temperature and the other maintained at 10C above. Maximum water temperat...

120

Multiple Scales in the Simulation of Ion Channels and Proteins  

PubMed Central

Computation of living processes creates great promise for the everyday life of mankind and great challenges for physical scientists. Simulations molecular dynamics have great appeal to biologists as a natural extension of structural biology. Once a biologist sees a structure, she/he wants to see it move. Molecular biology has shown that a small number of atoms, sometimes even one messenger ion, like Ca2+, can control biological function on the scale of cells, organs, tissues, and organisms. Enormously concentrated ions—at number densities of ~20 M—in protein channels and enzymes are responsible for many of the characteristics of living systems, just as highly concentrated ions near electrodes are responsible for many of the characteristics of electrochemical systems. Here we confront the reality of the scale differences of ions. We show that the scale differences needed to simulate all the atoms of biological cells are 107 in linear dimension, 1021 in three dimensions, 109 in resolution, 1011 in time, and 1013 in particle number (to deal with concentrations of Ca2+). These scales must be dealt with simultaneously if the simulation is to deal with most biological functions. Biological function extends across all of them, all at once in most cases. We suggest a computational approach using explicit multiscale analysis instead of implicit simulation of all scales. The approach is based on an energy variational principle EnVarA introduced by Chun Liu to deal with complex fluids. Variational methods deal automatically with multiple interacting components and scales. When an additional component is added to the system, the resulting Euler Lagrange field equations change form automatically—by algebra alone—without additional unknown parameters. Multifaceted interactions are solutions of the resulting equations. We suggest that ionic solutions should be viewed as complex fluids with simple components. Highly concentrated solutions—dominated by interactions of components—are easily computed by EnVarA. Successful computation of ions concentrated in special places may be a significant step to understanding the defining characteristics of biological and electrochemical systems. Indeed, computing ions near proteins and nucleic acids may prove as important to molecular biology and chemical technology as computing holes and electrons has been to our semiconductor and digital technology.

Eisenberg, Bob

2010-01-01

121

The effect of water on protein dynamics.  

PubMed

Neutron diffraction and spectroscopy were applied to describe the hydration and dynamics of a soluble protein and a natural membrane from extreme halophilic Archaea. The quantitative dependence of protein motions on water activity was clearly illustrated, and it was established that a minimum hydration shell is required for the systems to access their functional resilience, i.e. a dynamics state that allows biological activity. PMID:15306381

Zaccai, G

2004-08-29

122

Oligomerization state of water channels and glycerol facilitators. Involvement of loop E.  

PubMed

The major intrinsic protein (MIP) family includes water channels aquaporins (AQPs) and facilitators for small solutes such as glycerol (GlpFs). Velocity sedimentation on sucrose gradients demonstrates that heterologous AQPcic expressed in yeast or Xenopus oocytes behaves as an homotetramer when extracted by n-octyl beta-D-glucopyranoside (OG) and as a monomer when extracted by SDS. We performed an analysis of GlpF solubilized from membranes of Escherichia coli or of mRNA-injected Xenopus oocytes. The GlpF protein extracted either by SDS or by nondenaturing detergents, OG and Triton X-100, exhibits sedimentation coefficients only compatible with a monomeric form of the protein in micelles. We then substituted in loop E of AQPcic two amino acids predicted to play a role in the functional/structural properties of the MIPs. In two expression systems, yeast and oocytes, the mutant AQPcic-S205D is monomeric in OG and in SDS. The A209K mutation does not modify the tetrameric form of the heterologous protein in OG. This study shows that the serine residue at position 205 is essential for AQPcic tetramerization. Because the serine in this position is highly conserved among aquaporins and systematically replaced by an acid aspartic in GlpFs, we postulate that glycerol facilitators are monomers whereas aquaporins are organized in tetramers. Our data suggest that the role of loop E in MIP properties partly occurs through its ability to allow oligomerization of the proteins. PMID:9852047

Lagrée, V; Froger, A; Deschamps, S; Pellerin, I; Delamarche, C; Bonnec, G; Gouranton, J; Thomas, D; Hubert, J F

1998-12-18

123

Regulation of Chloride Channels by Protein Kinase C in Normal and Cystic Fibrosis Airway Epithelia  

NASA Astrophysics Data System (ADS)

Apical membrane chloride channels control chloride secretion by airway epithelial cells. Defective regulation of these channels is a prominent characteristic of cystic fibrosis. In normal intact cells, activation of protein kinase C (PKC) by phorbol ester either stimulated or inhibited chloride secretion, depending on the physiological status of the cell. In cell-free membrane patches, PKC also had a dual effect: at a high calcium concentration, PKC inactivated chloride channels; at a low calcium concentration, PKC activated chloride channels. In cystic fibrosis cells, PKC-dependent channel inactivation was normal, but activation was defective. Thus it appears that PKC phosphorylates and regulates two different sites on the channel or on an associated membrane protein, one of which is defective in cystic fibrosis.

Li, Ming; McCann, John D.; Anderson, Matthew P.; Clancy, John P.; Liedtke, Carole M.; Nairn, Angus C.; Greengard, Paul; Welsh, Michael J.

1989-06-01

124

Remodeling of Channel-Forming ORAI Proteins Determines an Oncogenic Switch in Prostate Cancer.  

PubMed

ORAI family channels have emerged as important players in malignant transformation, yet the way in which they reprogram cancer cells remains elusive. Here we show that the relative expression levels of ORAI proteins in prostate cancer are different from that in noncancerous tissue. By mimicking ORAI protein remodeling observed in primary tumors, we demonstrate in in vitro models that enhanced ORAI3 expression favors heteromerization with ORAI1 to form a novel channel. These channels support store-independent Ca(2+) entry, thereby promoting cell proliferation and a smaller number of functional homomeric ORAI1-based store-operated channels, which are important in supporting susceptibility to apoptosis. Thus, our findings highlight disrupted dynamic equilibrium of channel-forming proteins as an oncogenic mechanism. PMID:24954132

Dubois, Charlotte; Vanden Abeele, Fabien; Lehen'kyi, V'yacheslav; Gkika, Dimitra; Guarmit, Basma; Lepage, Gilbert; Slomianny, Christian; Borowiec, Anne Sophie; Bidaux, Gabriel; Benahmed, Mohamed; Shuba, Yaroslav; Prevarskaya, Natalia

2014-07-14

125

Cold-acclimation-induced protein hypertrophy in channel catfish and green sunfish  

Microsoft Academic Search

Summary 1.Following acclimation of channel catfish to a reduction in temperature from 25° to 15°C, there were approximately two-fold increases in liver mass, cell size, total protein, and total enzyme activity, relative to activity per milligram of protein and per gram wet weight of tissue, indicating tissue hypertrophy. There was no change in either total liver DNA content or protein

Jeffrey Kent; Michael Koban; C. Ladd Prosser

1988-01-01

126

Accessory Protein Facilitated CFTR-CFTR Interaction, a Molecular Mechanism to Potentiate the Chloride Channel Activity  

Microsoft Academic Search

The c ystic f ibrosis t ransmembrane conductance r egulator (CFTR) gene encodes a chloride channel protein that belongs to the superfamily of A TP b inding c assette (ABC) transporters. Phosphorylation by protein kinase A in the presence of ATP activates the CFTR-mediated chloride conductance of the apical membranes. We have identified a novel hydrophilic CFTR binding protein, CAP70,

Shusheng Wang; Hongwen Yue; Rachel B. Derin; William B. Guggino; Min Li

2000-01-01

127

Effects of Protein, Amino Acid Levels, and Feeding Methods on Growth of Fingerling Channel Catfish.  

National Technical Information Service (NTIS)

Survival of fingerling channel catfish was nearly identical for fish on diets containing 25 percent protein and 22 percent protein. Average gain per fish, gain per pond, and feed conversion were improved when fish received diets with 25 percent protein. L...

C. W. Deyoe O. W. Tiemeier C. Suppes

1968-01-01

128

Proton transfer via a transient linear water-molecule chain in a membrane protein  

PubMed Central

High-resolution protein ground-state structures of proton pumps and channels have revealed internal protein-bound water molecules. Their possible active involvement in protein function has recently come into focus. An illustration of the formation of a protonated protein-bound water cluster that is actively involved in proton transfer was described for the membrane protein bacteriorhodopsin (bR) [Garczarek F, Gerwert K (2006) Nature 439:109–112]. Here we show through a combination of time-resolved FTIR spectroscopy and molecular dynamics simulations that three protein-bound water molecules are rearranged by a protein conformational change that resulted in a transient Grotthuss-type proton-transfer chain extending through a hydrophobic protein region of bR. This transient linear water chain facilitates proton transfer at an intermediate conformation only, thereby directing proton transfer within the protein. The rearrangement of protein-bound water molecules that we describe, from inactive positions in the ground state to an active chain in an intermediate state, appears to be energetically favored relative to transient incorporation of water molecules from the bulk. Our discovery provides insight into proton-transfer mechanisms through hydrophobic core regions of ubiquitous membrane spanning proteins such as G-protein coupled receptors or cytochrome C oxidases.

Freier, Erik; Wolf, Steffen; Gerwert, Klaus

2011-01-01

129

Diffusion, molecular separation, and drug delivery from lipid mesophases with tunable water channels.  

PubMed

Lyotropic liquid crystals characterized by a bicontinuous cubic phase (BCP) have a structure characterized by interpenetrated water channels following triply periodic minimal surfaces, which can be stable in excess water conditions and thus suitable in a multitude of applications. The control of the water channels size in these systems has a direct impact on their use for drug delivery, crystallization, and membrane separation processes. In this work we carry out systematic diffusion studies to show how the control on the water channel dimensions directly correlates with the release and separation performance of bicontinuous cubic phases. Specifically, we tune the water channels diameter of the monolinolein/water system by adding different amounts of sucrose stearate, which, having hydration-enhancing properties, can shift the boundaries of the phase diagram. We then design a model bicontinuous cubic phase lipidic membrane of the Im3m space group, having a sugar ester to monolinolein ratio of 20%, and we follow the diffusion within its water channels, by using molecules that differ systematically in size and molecular conformation, and we demonstrate, for each class of molecules, a diffusion-enhanced process upon increase of the water channel diameter. Finally, we also show the ability of the bicontinuous cubic phase to efficiently and selectively separate nanoparticles of a target size, by choosing an amount of sucrose stearate for which the water channel diameter and the nanoparticle dimensions match, demonstrating the possible use of these systems as filtering membranes of tunable molecular cutoff. PMID:23116138

Negrini, Renata; Mezzenga, Raffaele

2012-11-27

130

Phycodnavirus Potassium Ion Channel Proteins Question the Virus Molecular Piracy Hypothesis  

PubMed Central

Phycodnaviruses are large dsDNA, algal-infecting viruses that encode many genes with homologs in prokaryotes and eukaryotes. Among the viral gene products are the smallest proteins known to form functional K+ channels. To determine if these viral K+ channels are the product of molecular piracy from their hosts, we compared the sequences of the K+ channel pore modules from seven phycodnaviruses to the K+ channels from Chlorella variabilis and Ectocarpus siliculosus, whose genomes have recently been sequenced. C. variabilis is the host for two of the viruses PBCV-1 and NY-2A and E. siliculosus is the host for the virus EsV-1. Systematic phylogenetic analyses consistently indicate that the viral K+ channels are not related to any lineage of the host channel homologs and that they are more closely related to each other than to their host homologs. A consensus sequence of the viral channels resembles a protein of unknown function from a proteobacterium. However, the bacterial protein lacks the consensus motif of all K+ channels and it does not form a functional channel in yeast, suggesting that the viral channels did not come from a proteobacterium. Collectively, our results indicate that the viruses did not acquire their K+ channel-encoding genes from their current algal hosts by gene transfer; thus alternative explanations are required. One possibility is that the viral genes arose from ancient organisms, which served as their hosts before the viruses developed their current host specificity. Alternatively the viral proteins could be the origin of K+ channels in algae and perhaps even all cellular organisms.

Hamacher, Kay; Greiner, Timo; Ogata, Hiroyuki; Van Etten, James L.; Gebhardt, Manuela; Villarreal, Luis P.; Cosentino, Cristian; Moroni, Anna; Thiel, Gerhard

2012-01-01

131

Phycodnavirus potassium ion channel proteins question the virus molecular piracy hypothesis.  

PubMed

Phycodnaviruses are large dsDNA, algal-infecting viruses that encode many genes with homologs in prokaryotes and eukaryotes. Among the viral gene products are the smallest proteins known to form functional K(+) channels. To determine if these viral K(+) channels are the product of molecular piracy from their hosts, we compared the sequences of the K(+) channel pore modules from seven phycodnaviruses to the K(+) channels from Chlorella variabilis and Ectocarpus siliculosus, whose genomes have recently been sequenced. C. variabilis is the host for two of the viruses PBCV-1 and NY-2A and E. siliculosus is the host for the virus EsV-1. Systematic phylogenetic analyses consistently indicate that the viral K(+) channels are not related to any lineage of the host channel homologs and that they are more closely related to each other than to their host homologs. A consensus sequence of the viral channels resembles a protein of unknown function from a proteobacterium. However, the bacterial protein lacks the consensus motif of all K(+) channels and it does not form a functional channel in yeast, suggesting that the viral channels did not come from a proteobacterium. Collectively, our results indicate that the viruses did not acquire their K(+) channel-encoding genes from their current algal hosts by gene transfer; thus alternative explanations are required. One possibility is that the viral genes arose from ancient organisms, which served as their hosts before the viruses developed their current host specificity. Alternatively the viral proteins could be the origin of K(+) channels in algae and perhaps even all cellular organisms. PMID:22685610

Hamacher, Kay; Greiner, Timo; Ogata, Hiroyuki; Van Etten, James L; Gebhardt, Manuela; Villarreal, Luis P; Cosentino, Cristian; Moroni, Anna; Thiel, Gerhard

2012-01-01

132

Primary structure and functional expression of a rat G-protein-coupled muscarinic potassium channel  

Microsoft Academic Search

PARASYMPATHETIC nerve stimulation causes slowing of the heart rate by activation of muscarinic receptors and the subsequent opening of muscarinic K+ channels in the sinoatrial node and atrium1-4. This inwardly rectifying K+ channel is coupled directly with G protein5-10. Based on sequence homology with cloned inwardly rectifying K+ channels, ROMK1 (ref. 11) and IRK1 (ref. 12), we have isolated a

Yoshihiro Kubo; Eitan Reuveny; Paul A. Slesinger; Yuh Nung Jan; Lily Y. Jan

1993-01-01

133

Nonlinear Data assimilation for shallow water equations in branched channels  

NASA Astrophysics Data System (ADS)

A framework has been developed for the optimal combination of a network of time series observations with a numerical model of nonlinear shallow water wave propagation in branched channels. The model results and observations are weighted according to their reliability to produce estimates of surface elevation and transport. Two filtering techniques are adopted to perform the dynamic-stochastic modeling. For weakly nonlinear conditions a modified incremental covariance Chandrasekhar-type algorithm is employed, whereas for more strongly nonlinear dynamics a Bierman square root form of the extended Kalman filter is used. Numerical experiments performed using parameters from the Great Bay estuary in New Hampshire demonstrate that both filters successfully estimate the time- and space-dependent elevation and transport distributions under conditions of stochastic forcing (model error) and measurement noise. Although the Chandrasekhar algorithm underestimates the covariances of the filter estimates, it performs nearly as well as the fully nonlinear algorithm, even for the significantly nonlinear test case. It is shown that the Chandrasekhar filter performance does not degrade when the algorithm is applied to systems in which the process noise and measurement noise structures are purely sinusoidal, thus violating the assumption of white noise under which the filter algorithm is derived.

Budgell, W. P.

1986-09-01

134

Ferritin Protein Nanocages Use Ion Channels, Catalytic Sites, and Nucleation Channels To Manage Iron/Oxygen Chemistry  

PubMed Central

The ferritin superfamily is composed of ancient, nanocage proteins with an internal cavity, 60% of total volume, that reversibly synthesize solid minerals of hydrated ferric oxide; the minerals are iron concentrates for cell nutrition as well as antioxidants due to ferrous and oxygen consumption during mineralization. The cages have multiple iron entry/exit channels, oxidoreductase enzyme sites, and, in eukaryotes, Fe(III)O nucleation channels with clustered exits that extend protein activity to include facilitated mineral growth. Ferritin protein cage differences include size, amino acid sequence, and location of the active sites, oxidant substrate and crystallinity of the iron mineral. Genetic regulation depends on iron and oxygen signals, which in animals includes direct ferrous signaling to RNA to release and to ubiquitin-ligases to degrade the protein repressors. Ferritin biosynthesis forms, with DNA, mRNA and the protein product, a feedback loop where the genetic signals are also protein substrates. The ferritin protein nanocages, which are required for normal iron homeostasis and are finding current use in delivery of nanodrugs, novel nanomaterials, and nanocatalysts, are likely contributors to survival and success during the transition from anaerobic to aerobic life.

Theil, Elizabeth C.

2011-01-01

135

Voltage Gated Sodium Channel Associated Proteins and Alternative Mechanisms of Inactivation and Block  

PubMed Central

Voltage-gated sodium channels mediate inward current of action potentials upon membrane depolarization of excitable cells. The initial transient sodium current is restricted to milliseconds through three distinct channel inactivating and blocking mechanisms. All pore-forming alpha subunits of sodium channels possess structural elements mediating fast inactivation upon depolarization and recovery within milliseconds upon membrane repolarization. Accessory subunits modulate fast inactivation dynamics, but these proteins can also limit current by contributing distinct inactivation and blocking particles. A-type isoforms of fibroblast growth factor homologous factors (FHFs) bear a particle that induces long-term channel inactivation, while sodium channel subunit Nav?4 employs a blocking particle that rapidly dissociates upon membrane repolarization to generate resurgent current. Despite their different physiological functions, the FHF and Nav?4 particles have similarity in amino acid composition and mechanisms for docking within sodium channels. The three competing channel inactivating and blocking processes functionally interact to regulate a neuron’s intrinsic excitability.

Goldfarb, Mitchell

2011-01-01

136

The ABC protein turned chloride channel whose failure causes cystic fibrosis  

Microsoft Academic Search

CFTR chloride channels are encoded by the gene mutated in patients with cystic fibrosis. These channels belong to the superfamily of ABC transporter ATPases. ATP-driven conformational changes, which in other ABC proteins fuel uphill substrate transport across cellular membranes, in CFTR open and close a gate to allow transmembrane flow of anions down their electrochemical gradient. New structural and biochemical

David C. Gadsby; Paola Vergani; László Csanády

2006-01-01

137

The jellyfish green fluorescent protein: A new tool for studying ion channel expression and function  

Microsoft Academic Search

Two methods are described for using the jellyfish green fluorescent protein (GFP) as a reporter gene for ion channel expression. GFP fluorescence can be used to identify the transfected cells, and to estimate the relative levels of ion channel expression, in cotransfection experiments. A GFP-NMDAR1 chimera can be constructed that produces a functional, fluorescent receptor subunit. These methods should facilitate

John Marshall; Raymond Molloy; Guy W. J Moss; James R Howe; Thomas E Hughes

1995-01-01

138

Slow modal gating of single G protein-activated K+ channels expressed in Xenopus oocytes  

PubMed Central

The slow kinetics of G protein-activated K+ (GIRK) channels expressed in Xenopus oocytes were studied in single-channel, inside-out membrane patches. Channels formed by GIRK1 plus GIRK4 subunits, which are known to form the cardiac acetylcholine (ACh)-activated GIRK channel (KACh), were activated by a near-saturating dose of G protein ?? subunits (G??; 20 nM). The kinetic parameters of the expressed GIRK1/4 channels were similar to those of cardiac KACh. GIRK1/4 channels differed significantly from channels formed by GIRK1 with the endogenous oocyte subunit GIRK5 (GIRK1/5) in some of their kinetic parameters and in a 3-fold lower open probability, Po. The unexpectedly low Po (0.025) of GIRK1/4 was due to the presence of closures of hundreds of milliseconds; the channel spent ?90 % of the time in the long closed states. GIRK1?4 channels displayed a clear modal behaviour: on a time scale of tens of seconds, the G??-activated channels cycled between a low-Po mode (Po of about 0.0034) and a bursting mode characterized by an ?30-fold higher Po and a different set of kinetic constants (and, therefore, a different set of channel conformations). The available evidence indicates that the slow modal transitions are not driven by binding and unbinding of G??. The GTP?S-activated G?i1 subunit, previously shown to inhibit GIRK channels, substantially increased the time spent in closed states and apparently shifted the channel to a mode similar, but not identical, to the low-Po mode. This is the first demonstration of slow modal transitions in GIRK channels. The detailed description of the slow gating kinetics of GIRK1?4 may help in future analysis of mechanisms of GIRK gating.

Yakubovich, Daniel; Pastushenko, Vassili; Bitler, Arkadi; Dessauer, Carmen W; Dascal, Nathan

2000-01-01

139

Effect of Heavy Water on Protein Flexibility  

Microsoft Academic Search

The effects of heavy water (D2O) on internal dynamics of proteins were assessed by both the intrinsic phosphorescence lifetime of deeply buried Trp residues, which reports on the local structure about the triplet probe, and the bimolecular acrylamide phosphorescence quenching rate constant that is a measure of the average acrylamide diffusion coefficient through the macromolecule. The results obtained with several

Patrizia Cioni; Giovanni B. Strambini

2002-01-01

140

Bioluminescence Methodology for the Detection of Protein-Protein Interactions Within the Voltage-Gated Sodium Channel Macromolecular Complex  

PubMed Central

Abstract Protein–protein interactions are critical molecular determinants of ion channel function and emerging targets for pharmacological interventions. Yet, current methodologies for the rapid detection of ion channel macromolecular complexes are still lacking. In this study we have adapted a split-luciferase complementation assay (LCA) for detecting the assembly of the voltage-gated Na+ (Nav) channel C-tail and the intracellular fibroblast growth factor 14 (FGF14), a functionally relevant component of the Nav channelosome that controls gating and targeting of Nav channels through direct interaction with the channel C-tail. In the LCA, two complementary N-terminus and C-terminus fragments of the firefly luciferase were fused, respectively, to a chimera of the CD4 transmembrane segment and the C-tail of Nav1.6 channel (CD4-Nav1.6-NLuc) or FGF14 (CLuc-FGF14). Co-expression of CLuc-FGF14 and CD4-Nav1.6-NLuc in live cells led to a robust assembly of the FGF14:Nav1.6 C-tail complex, which was attenuated by introducing single-point mutations at the predicted FGF14:Nav channel interface. To evaluate the dynamic regulation of the FGF14:Nav1.6 C-tail complex by signaling pathways, we investigated the effect of kinase inhibitors on the complex formation. Through a platform of counter screenings, we show that the p38/MAPK inhibitor, PD169316, and the I?B kinase inhibitor, BAY 11-7082, reduce the FGF14:Nav1.6 C-tail complementation, highlighting a potential role of the p38MAPK and the I?B/NF?B pathways in controlling neuronal excitability through protein–protein interactions. We envision the methodology presented here as a new valuable tool to allow functional evaluations of protein–channel complexes toward probe development and drug discovery targeting ion channels implicated in human disorders.

Shavkunov, Alexander; Panova, Neli; Prasai, Anesh; Veselenak, Ron; Bourne, Nigel; Stoilova-McPhie, Svetla

2012-01-01

141

Ancient association between cation leak channels and Mid1 proteins is conserved in fungi and animals  

PubMed Central

Neuronal resting potential can tune the excitability of neural networks, affecting downstream behavior. Sodium leak channels (NALCN) play a key role in rhythmic behaviors by helping set, or subtly changing neuronal resting potential. The full complexity of these newly described channels is just beginning to be appreciated, however. NALCN channels can associate with numerous subunits in different tissues and can be activated by several different peptides and second messengers. We recently showed that NALCN channels are closely related to fungal calcium channels, which they functionally resemble. Here, we use this relationship to predict a family of NALCN-associated proteins in animals on the basis of homology with the yeast protein Mid1, the subunit of the yeast calcium channel. These proteins all share a cysteine-rich region that is necessary for Mid1 function in yeast. We validate this predicted association by showing that the Mid1 homolog in Drosophila, encoded by the CG33988 gene, is coordinately expressed with NALCN, and that knockdown of either protein creates identical phenotypes in several behaviors associated with NALCN function. The relationship between Mid1 and leak channels has therefore persisted over a billion years of evolution, despite drastic changes to both proteins and the organisms in which they exist.

Ghezzi, Alfredo; Liebeskind, Benjamin J.; Thompson, Ammon; Atkinson, Nigel S.; Zakon, Harold H.

2014-01-01

142

Investigation of water droplet dynamics in PEM fuel cell gas channels  

NASA Astrophysics Data System (ADS)

Water management in Proton Exchange Membrane Fuel Cell (PEMFC) has remained one of the most important issues that need to be addressed before its commercialization in automotive applications. Accumulation of water on the gas diffusion layer (GDL) surface in a PEMFC introduces a barrier for transport of reactant gases through the GDL to the catalyst layer. Despite the fact that the channel geometry is one of the key design parameters of a fluidic system, very limited research is available to study the effect of microchannel geometry on the two-phase flow structure. In this study, the droplet-wall dynamics and two-phase pressure drop across the water droplet present in a typical PEMFC channel, were examined in auto-competitive gas channel designs (0.4 x 0.7 mm channel cross section). The liquid water flow pattern inside the gas channel was analyzed for different air velocities. Experimental data was analyzed using the Concus-Finn condition to determine the wettability characteristics in the corner region. It was confirmed that the channel angle along with the air velocity and the channel material influences the water distribution and holdup within the channel. Dynamic contact angle emerged as an important parameter in controlling the droplet-wall interaction. Experiments were also performed to understand how the inlet location of the liquid droplet on the GDL surface affects the droplet dynamic behavior in the system. It was found that droplets emerging near the channel wall or under the land lead to corner filling of the channel. Improvements in the channel design has been proposed based on the artificial channel roughness created to act as capillary grooves to transport the liquid water away from the land area. For droplets emerging near the center of the channel, beside the filling and no-filling behavior reported in the literature, a new droplet jumping behavior was observed. As droplets grew and touched the sidewalls, they jumped off to the sidewall leaving the whole GDL exposed for gases to diffuse to the catalyst layer. A theoretical model was developed and a criterion was proposed to predict the droplet jumping behavior in the gas channel. A theoretical force balance model was proposed to predict the pressure force and air velocity required to remove the droplet from the channel to avoid complete channel blockage. The overall goal of this work was to identify the gas channel configuration that provides efficient water removal with a lower pressure drop in the system efficiency while meeting the US Department of Energy's specifications for a PEMFC for automotive application.

Gopalan, Preethi

143

Lens cell-to-cel channel protein: II. Conformational change in the presence of calmodulin  

Microsoft Academic Search

Summary Lens fibers are coupled by communicating junctions, clusters of cell-to-cell channels composed of a 28-kD intrinsic membrane protein (MIP26). Evidence suggests that these and other cell-to-cell channels may close as a result of protein conformational change induced by activated calmodulin. To test the validity of this hypothesis, we have measured the intrinsic fluorescence emission and far-ultraviolet circular dichroism of

S. J. Girsch; C. Peracchia

1985-01-01

144

Role of Derlin-1 protein in proteostasis regulation of ATP-sensitive potassium channels.  

PubMed

ATP-sensitive potassium (K(ATP)) channels composed of sulfonylurea receptor 1 (SUR1) and Kir6.2 regulate insulin secretion by linking glucose metabolism with membrane potential. The number of K(ATP) channels in the plasma membrane affects the sensitivity of ?-cells to glucose. Aberrant surface channel expression leads to insulin secretion disease. Previously, we have shown that K(ATP) channel proteins undergo endoplasmic reticulum (ER)-associated degradation (ERAD) via the ubiquitin-proteasome pathway, and inhibition of proteasome function results in an increase in channel surface expression. Here, we investigated whether Derlin-1, a protein involved in retrotranslocation of misfolded or misassembled proteins across the ER membrane for degradation by cytosolic proteasomes, plays a role in ERAD and, in turn, biogenesis efficiency of K(ATP) channels. We show that both SUR1 and Kir6.2 form a complex with Derlin-1 and an associated AAA-ATPase, p97. Overexpression of Derlin-1 led to a decrease in the biogenesis efficiency and surface expression of K(ATP) channels. Conversely, knockdown of Derlin-1 by RNA interference resulted in increased processing of SUR1 and a corresponding increase in surface expression of K(ATP) channels. Importantly, knockdown of Derlin-1 increased the abundance of disease-causing misfolded SUR1 or Kir6.2 proteins and even partially rescued surface expression in a mutant channel. We conclude that Derlin-1, by being involved in ERAD of SUR1 and Kir6.2, has a role in modulating the biogenesis efficiency and surface expression of K(ATP) channels. The results suggest that physiological or pathological changes in Derlin-1 expression levels may affect glucose-stimulated insulin secretion by altering surface expression of K(ATP) channels. PMID:22311976

Wang, Fang; Olson, Erik M; Shyng, Show-Ling

2012-03-23

145

CORONAVIRUS E PROTEIN FORMS ION CHANNELS WITH FUNCTIONALLY AND STRUCTURALLY-INVOLVED MEMBRANE LIPIDS  

PubMed Central

Coronavirus (CoV) envelope (E) protein ion channel activity was determined in channels formed in planar lipid bilayers by peptides representing either the transmembrane domain of severe acute respiratory syndrome CoV (SARS-CoV) E protein, or the full-length E protein. Both of them formed a voltage independent ion conductive pore with symmetric ion transport properties. Mutations N15A and V25F located in the transmembrane domain, prevented the ion conductivity. E protein derived channels showed no cation preference in non-charged lipid membranes, whereas they behaved as pores with mild cation selectivity in negatively-charged lipid membranes. The ion conductance was also controlled by the lipid composition of the membrane. Lipid charge also regulated the selectivity of a HCoV-229E E protein derived peptide. These results suggested that the lipids are functionally involved in E protein ion channel activity, forming a protein-lipid pore, a novel concept for CoV E protein ion channel entity.

Verdia-Baguena, Carmina; Nieto-Torres, Jose L.; Alcaraz, Antonio; DeDiego, Marta L.; Torres, Jaume; Aguilella, Vicente M.; Enjuanes, Luis

2012-01-01

146

The ABC protein turned chloride channel whose failure causes cystic fibrosis  

NASA Astrophysics Data System (ADS)

CFTR chloride channels are encoded by the gene mutated in patients with cystic fibrosis. These channels belong to the superfamily of ABC transporter ATPases. ATP-driven conformational changes, which in other ABC proteins fuel uphill substrate transport across cellular membranes, in CFTR open and close a gate to allow transmembrane flow of anions down their electrochemical gradient. New structural and biochemical information from prokaryotic ABC proteins and functional information from CFTR channels has led to a unifying mechanism explaining those ATP-driven conformational changes.

Gadsby, David C.; Vergani, Paola; Csanády, László

2006-03-01

147

A novel mouse Nedd4 protein suppresses the activity of the epithelial Na+ channel  

Microsoft Academic Search

Liddle's syndrome is a form of inherited hypertension linked to mutations in the genes encoding the epithelial Na1 channel (ENaC). These mutations alter or delete PY motifs involved in protein-protein interactions with a ubiquitin-protein ligase, Nedd4. Here we show that Na1 transporting cells, derived from mouse cortical collecting duct, express two Nedd4 proteins with different structural organization and characteristics of

ELENA KAMYNINA; CHRISTOPHE DEBONNEVILLE; MARCELLE BENS; ALAIN VANDEWALLE; OLIVIER STAUB

2001-01-01

148

Aquaporin-4 water channels and synaptic plasticity in the hippocampus.  

PubMed

Aquaporin-4 (AQP4) is the major water channel expressed in the central nervous system (CNS) and is primarily expressed in glial cells. Many studies have shown that AQP4 regulates the response of the CNS to insults or injury, but far less is known about the potential for AQP4 to influence synaptic plasticity or behavior. Recent studies have examined long-term potentiation (LTP), long-term depression (LTD), and behavior in AQP4 knockout (KO) and wild-type mice to gain more insight into its potential role. The results showed a selective effect of AQP4 deletion on LTP of the Schaffer collateral pathway in hippocampus using an LTP induction protocol that simulates pyramidal cell firing during theta oscillations (theta-burst stimulation; TBS). However, LTP produced by a different induction protocol was unaffected. There was also a defect in LTD after low frequency stimulation (LFS) in AQP4 KO mice. Interestingly, some slices from AQP4 KO mice exhibited LTD after TBS instead of LTP, or LTP following LFS instead of LTD. These data suggest that AQP4 and astrocytes influence the polarity of long-term synaptic plasticity (potentiation or depression). These potentially powerful roles expand the influence of AQP4 and astrocytes beyond the original suggestions related to regulation of extracellular potassium and water balance. Remarkably, AQP4 KO mice did not show deficits in basal transmission, suggesting specificity for long-term synaptic plasticity. The mechanism appears to be related to neurotrophins and specifically brain-derived neurotrophic factor (BDNF) because pharmacological blockade of neurotrophin trk receptors or scavenging ligands such as BDNF restored plasticity. The in vitro studies predicted effects in vivo of AQP4 deletion because AQP4 KO mice performed worse using a task that requires memory for the location of objects (object placement). However, performance on other hippocampal-dependent tasks was spared. The results suggest an unanticipated and selective role of AQP4 in synaptic plasticity and spatial memory, and underscore the growing appreciation of the role of glial cells in functions typically attributed to neurons. Implications for epilepsy are discussed because of the previous evidence that AQP4 influences seizures, and the role of synaptic plasticity in epileptogenesis. PMID:23684954

Scharfman, Helen E; Binder, Devin K

2013-12-01

149

Akt regulates L-type Ca2+ channel activity by modulating Cav?1 protein stability  

PubMed Central

The insulin IGF-1–PI3K–Akt signaling pathway has been suggested to improve cardiac inotropism and increase Ca2+ handling through the effects of the protein kinase Akt. However, the underlying molecular mechanisms remain largely unknown. In this study, we provide evidence for an unanticipated regulatory function of Akt controlling L-type Ca2+ channel (LTCC) protein density. The pore-forming channel subunit Cav?1 contains highly conserved PEST sequences (signals for rapid protein degradation), and in-frame deletion of these PEST sequences results in increased Cav?1 protein levels. Our findings show that Akt-dependent phosphorylation of Cav?2, the LTCC chaperone for Cav?1, antagonizes Cav?1 protein degradation by preventing Cav?1 PEST sequence recognition, leading to increased LTCC density and the consequent modulation of Ca2+ channel function. This novel mechanism by which Akt modulates LTCC stability could profoundly influence cardiac myocyte Ca2+ entry, Ca2+ handling, and contractility.

Zhang, Deng-Hong; DeSantiago, Jaime; Aimond, Franck; Barbara, Guillaume; Chemin, Jean; Bonci, Desire; Picht, Eckard; Rusconi, Francesca; Dalton, Nancy D.; Peterson, Kirk L.; Richard, Sylvain; Bers, Donald M.; Brown, Joan Heller; Condorelli, Gianluigi

2009-01-01

150

G Protein ?? Gating Confers Volatile Anesthetic Inhibition to Kir3 Channels*  

PubMed Central

G protein-activated inwardly rectifying potassium (GIRK or Kir3) channels are directly gated by the ?? subunits of G proteins and contribute to inhibitory neurotransmitter signaling pathways. Paradoxically, volatile anesthetics such as halothane inhibit these channels. We find that neuronal Kir3 currents are highly sensitive to inhibition by halothane. Given that Kir3 currents result from increased G?? available to the channels, we asked whether reducing available G?? to the channel would adversely affect halothane inhibition. Remarkably, scavenging G?? using the C-terminal domain of ?-adrenergic receptor kinase (c?ARK) resulted in channel activation by halothane. Consistent with this effect, channel mutants that impair G?? activation were also activated by halothane. A single residue, phenylalanine 192, occupies the putative G?? gate of neuronal Kir3.2 channels. Mutation of Phe-192 at the gate to other residues rendered the channel non-responsive, either activated or inhibited by halothane. These data indicated that halothane predominantly interferes with G??-mediated Kir3 currents, such as those functioning during inhibitory synaptic activity. Our report identifies the molecular correlate for anesthetic inhibition of Kir3 channels and highlights the significance of these effects in modulating neurotransmitter-mediated inhibitory signaling.

Styer, Amanda M.; Mirshahi, Uyenlinh L.; Wang, Chuan; Girard, Laura; Jin, Taihao; Logothetis, Diomedes E.; Mirshahi, Tooraj

2010-01-01

151

Structure and chromosomal localization of a human water channel (AQP3) gene.  

PubMed

A cDNA encoding rat AQP3, a water channel and a member of the MIP family, that is expressed predominantly in kidney medulla and colon was cloned recently. To determine the structure, tissue distribution, and chromosomal localization of the human AQP3 gene, we screened a human kidney cDNA library with rat AQP3 probe and isolated a cDNA coding for human AQP3 protein. The deduced amino acid sequence of human AQP3 was 91% identical to rat AQP3. Human AQP3 mRNA was expressed in colon, kidney, liver, pancreas, lung, peripheral leukocytes, spleen, and prostate. The human AQP3 gene was mapped to 7q36.2-q36.3 by chromosome fluorescence in situ hybridization. PMID:7558005

Ishibashi, K; Sasaki, S; Saito, F; Ikeuchi, T; Marumo, F

1995-05-20

152

Structure and chromosomal localization of a human water channel (AQP3) gene  

SciTech Connect

A cDNA encoding rat AQP3, a water channel and a member of the MIP family, that is expressed predominantly in kidney medulla and colon was cloned recently. To determine the structure, tissue distribution, and chromosomal localization of the human AQP3 gene, the authors screened a human kidney cDNA library with rat AQP3 probe and isolated a cDNA coding for human AQP3 protein. The deduced amino acid sequence of human AQP3 was 91% identical to rat AQP3. Human AQP3 mRNA was expressed in colon, kidney, liver, pancreas, lung, peripheral leukocytes, spleen, and prostate. The human AQP3 gene was mapped to 7q36.2-q36.3 by chromosome fluorescence in situ hybridization. 10 refs., 3 figs.

Ishibashi, Kenichi; Sasaki, Sei; Saito, Fumiko [Tokyo Medical and Dental Univ. (Japan)] [and others] [Tokyo Medical and Dental Univ. (Japan); and others

1995-05-20

153

Monte Carlo simulation of the water in a channel with charges.  

PubMed Central

A Monte Carlo simulation of water in a channel with charges suggests the existence of water in immobile, high density, essentially glasslike form near the charges. The channel model has a conical section with an opening through which water molecules can pass, at the narrow end of the cone, and a cylindrical section at the other end. When the charges are placed near the narrow section of the model, the "glass" effectively blocks the channel; with the charges removed, the channel opens. The effect can be determined from the rate of passage of the water molecules through the pore, from the average orientation of the water molecule, and from distortion of the distribution of molecules. In the simulations carried out to date, no external ions have been considered. In addition to the energy, the Helmholtz free energy has been calculated.

Green, M E; Lewis, J

1991-01-01

154

Ion channel activity of brain abundant protein BASP1 in planar lipid bilayers.  

PubMed

BASP1 (also known as CAP-23 and NAP-22) is a brain abundant myristoylated protein localized at the inner surface of the presynaptic plasma membrane. Emerging evidence suggests that BASP1 is critically involved in various cellular processes, in particular, in the accumulation of phosphatidylinositol-4,5-diphosphate (PIP(2)) in lipid raft microdomains. We have recently shown that BASP1 forms heterogeneously-sized oligomers and higher aggregates with an outward similarity to oligomers and protofibrils of amyloid proteins. However, BASP1 is not known to be related to any amyloid disease. In the present study, we show that BASP1 induces single channel currents across negatively-charged planar lipid bilayers (containing phosphatidylserine or PIP(2)) bathed in 0.1-0.2 M KCl (pH 7.5). By their characteristics, BASP1 channels are similar to amyloid protein channels. BASP1 channels exhibit multiple conductance levels, in the range 10-3000 pS, with the most frequently observed conductance state of approximately 50 pS. The channels demonstrate a linear current-voltage relationship and voltage-independent kinetics of opening and closing. Their K(+) to Cl(-) permeability ratio is approximately 14, indicating that BASP1 channels are cation-selective. The ion channel activity of BASP1 is in accordance with the pore-like structure of BASP1 oligomers observed by electron microscopy on a lipid monolayer. Neuronal protein GAP-43, which is functionally related to BASP1 and also forms oligomers, elicited no ion channel currents under the conditions used in the present study. Elucidation of the physiological or pathological roles of ion channel activity of membrane-bound BASP1 oligomers will help to define the precise mechanism of amyloid protein toxicity. PMID:21156029

Ostroumova, Olga S; Schagina, Ludmila V; Mosevitsky, Mark I; Zakharov, Vladislav V

2011-02-01

155

76 FR 47529 - Port Access Route Study: In the Waters of Montauk Channel and Block Island Sound  

Federal Register 2010, 2011, 2012, 2013

...Waters of Montauk Channel and Block Island Sound AGENCY: Coast Guard, DHS. ACTION: Notice...Waters of Montauk Channel and Block Island Sound. The goals of the study are to help reduce...Waters of Montauk Channel and Block Island Sound. All comments received will be...

2011-08-05

156

Particularities of the dynamics of river channel processes in the Khabarovsk water junction  

Microsoft Academic Search

The dynamics of river channel processes were analyzed in the region of complex floodplain-channel branching of the Amur River\\u000a near Khabarovsk using remote sensing data of the Earth’s surface. Zones of steady accumulation and long-term erosion of the\\u000a river channel are established within the Khabarovsk water junction over the period of 1964–2004. The main problems related\\u000a to restoring the Amur

V. I. Sinyukov; V. A. Glukhov; O. V. Rybas; A. N. Makhinov; V. I. Kim; N. V. Berdnikov

2008-01-01

157

Cl Channels in CF: Lack of Activation by Protein Kinase C and cAMP-Dependent Protein Kinase  

Microsoft Academic Search

Secretory chloride channels can be activated by adenosine 3',5'-monophosphate (cAMP)-dependent protein kinase in normal airway epithelial cells but not in cells from individuals with cystic fibrosis (CF). In excised, inside-out patches of apical membrane of normal human airway cells and airway cells from three patients with CF, the chloride channels exhibited a characteristic outwardly rectifying current-voltage relation and depolarization-induced activation.

Tzyh-Chang Hwang; Luo Lu; Pamela L. Zeitlin; Dieter C. Gruenert; Richard Huganir; William B. Guggino

1989-01-01

158

Efficient water-filling algorithms for a Gaussian multiaccess channel with ISI  

Microsoft Academic Search

Although the optimum transmit power spectral density (PSD) of each user in a Gaussian multiaccess channel is theoretically available, efficient algorithms are needed to obtain the optimal PSD numerically. This paper presents two water-filling algorithms for the scenario where all users have the same priorities. First we propose for a two-user channel an exact algorithm with a computational complexity of

Chaohuang Zeng; Louise M. C. Hoo; John M. Cioffi

2000-01-01

159

Multicriteria decision analysis in water resources management: the malnichara channel improvement  

Microsoft Academic Search

Multicriteria decision analysis tool is used in many water resources and environmental management projects. The Malnichara is one of the natural channels in Sylhet city (Bangladesh) responsible for storm runoff conveyance to the downstream Surma river. The channel is found to be encroached at many locations of the city and found to be very vulnerable. The authority has taken decision

R. K. Chowdhury; R. Rahman

2008-01-01

160

Nano-environmental changes by KCNE proteins modify KCNQ channel function.  

PubMed

The KCNQ1 channel is a voltage-dependent potassium channel, which is widely expressed in various tissues of the human body including heart, inner ear, intestine, kidney and pancreas.  The ion channel properties of KCNQ1 change remarkably when auxiliary subunit KCNE proteins co-exist.  The mechanisms of KCNQ1 channel regulation by KCNE proteins are of longstanding interest but are still far from being fully understood.  The pore region (S5-S6 segments) of KCNQ1 is thought to be the main interaction site for KCNE proteins.  However, some recent reports showed that the voltage-sensing domain (S1-S4 segments) is critically involved in the regulation of KCNQ1 by KCNE proteins.  In addition, we recently re-examined the stoichiometry of the KCNQ1-KCNE1 complex and found that the stoichiometry is not fixed but rather flexible and the KCNQ1 channel can have up to four associated KCNE1 proteins.  We will review these recent findings concerning the mechanisms of KCNQ1 regulation by KCNE proteins. PMID:21654200

Nakajo, Koichi; Kubo, Yoshihiro

2011-01-01

161

The Major Outer Membrane Protein of Chlamydia psittaci Functions as a Porin-Like Ion Channel  

Microsoft Academic Search

The major outer membrane protein (MOMP) of Chlamydia species shares several biochemical properties with classical porin proteins. Secondary structure analysis by circular dichroism now reveals that MOMP purified from Chlamydia psittaci has a predominantly b-sheet content (62%), which is also typical of bacterial porins. Can MOMP form functional ion channels? To directly test the \\

SUSAN WYLLIE; RICHARD H. ASHLEY; DAVID LONGBOTTOM; ALAN J. HERRING

1998-01-01

162

A fast in silico simulation of ion flux through the large-pore channel proteins.  

PubMed Central

The PSST program (see accompanying article) utilizes the detailed structure of a large-pore channel protein as the sole input for selection of trajectories along which negative and positive ions propagate. In the present study we applied this program to reconstruct the ion flux through five large-pore channel proteins (PhoE, OmpF, the WT R. blastica general diffusion porin and two of its mutants). The conducting trajectories, one for positive and one for negative particles, are contorted pathways that run close to arrays of charged residues on the inner surface of the channel. In silico propagation of the charged particles yielded passage time values that are compatible with the measured average passage time of ions. The calculated ionic mobilities are close to those of the electrolyte solution of comparable concentrations. Inspection of the transition probabilities along the channel revealed no region that could impose a rate-limiting step. It is concluded that the ion flux is a function of the whole array of local barriers. Thus, the conductance of the large-pore channel protein is determined by the channel's shape and charge distribution, while the selectivity also reflects the features of the channel's vestibule.

Bransburg-Zabary, Sharron; Nachliel, Esther; Gutman, Menachem

2002-01-01

163

Gabapentin activates ROMK1 channels by a protein kinase A (PKA)-dependent mechanism  

PubMed Central

Background and purpose: Gabapentin is an effective anticonvulsant. The major physiological function of renal outer medullary potassium (ROMK1) channels is to maintain the resting membrane potential (RMP). We investigated the effect of gabapentin on ROMK1 channels and the mechanism involved. Experimental approach: Xenopus oocytes were injected with mRNA coding for wild-type or mutant ROMK1 channels and giant inside-out patch-clamp recordings were performed. Key results: Gabapentin increased the activity of ROMK1 channels, concentration-dependently and enhanced the activity of wild-type and an intracellular pH (pHi)-gating residue mutant (K80M) channels over a range of pHi. Gabapentin also increased activity of channels mutated at phosphatidylinositol 4,5-bisphosphate (PIP2)-binding sites (R188Q, R217A and K218A). However, gabapentin failed to enhance channel activity in the presence of protein kinase A (PKA) inhibitors and did not activate phosphorylation site mutants (S44A, S219A or S313A), mutants that mimicked the negative charge carried by a phosphate group bound to a serine (S44D, S219D or S313D), or a mutated channel with a positive charge (S219R). These findings show that gabapentin activates ROMK1 channels independently of the pHi and not via a PIP2-dependent pathway. The effects of gabapentin on ROMK1 channels may be due to a PKA-mediated phosphorylation-induced conformational change, but not to charge–charge interactions. Conclusions and implications: ROMK1 channels are the main channels responsible for maintaining the RMP during cellular excitation. Gabapentin increased the activity of ROMK1 channels by a PKA-dependent mechanism, reducing neuronal excitability, and this may play an important role in its antiepileptic effect.

Lee, C-H; Tsai, T-S; Liou, H-H

2008-01-01

164

Modulation of skeletal muscle sodium channels by human myotonin protein kinase.  

PubMed Central

In myotonic muscular dystrophy, abnormal muscle Na currents underlie myotonic discharges. Since the myotonic muscular dystrophy gene encodes a product, human myotonin protein kinase, with structural similarity to protein kinases, we tested the idea that human myotonin protein kinase modulates skeletal muscle Na channels. Coexpression of human myotonin protein kinase with rat skeletal muscle Na channels in Xenopus oocytes reduced the amplitude of Na currents and accelerated current decay. The effect required the presence of a potential phosphorylation site in the inactivation mechanism of the channel. The mutation responsible for human disease, trinucleotide repeats in the 3' untranslated region, did not prevent the effect. The consequence of an abnormal amount of the kinase would be altered muscle cell excitability, consistent with the clinical finding of myotonia in myotonic dystrophy. Images

Mounsey, J P; Xu, P; John, J E; Horne, L T; Gilbert, J; Roses, A D; Moorman, J R

1995-01-01

165

Influenza B virus BM2 protein has ion channel activity that conducts protons across membranes.  

PubMed

Successful uncoating of the influenza B virus in endosomes is predicted to require acidification of the interior of the virus particle. We report that a virion component, the BM2 integral membrane protein, when expressed in Xenopus oocytes or in mammalian cells, causes acidification of the cells and possesses ion channel activity consistent with proton conduction. Furthermore, coexpression of BM2 with hemagglutinin (HA) glycoprotein prevents HA from adopting its low-pH-induced conformation during transport to the cell surface, and overexpression of BM2 causes a delay in intracellular transport in the exocytic pathway and causes morphological changes in the Golgi. These data are consistent with BM2 equilibrating the pH gradient between the Golgi and the cytoplasm. The transmembrane domain of BM2 protein and the influenza A virus A/M2 ion channel protein both contain the motif HXXXW, and, for both proteins, the His and Trp residues are important for channel function. PMID:12852861

Mould, Jorgen A; Paterson, Reay G; Takeda, Makoto; Ohigashi, Yuki; Venkataraman, Padma; Lamb, Robert A; Pinto, Lawrence H

2003-07-01

166

Modulation of skeletal muscle sodium channels by human myotonin protein kinase.  

PubMed

In myotonic muscular dystrophy, abnormal muscle Na currents underlie myotonic discharges. Since the myotonic muscular dystrophy gene encodes a product, human myotonin protein kinase, with structural similarity to protein kinases, we tested the idea that human myotonin protein kinase modulates skeletal muscle Na channels. Coexpression of human myotonin protein kinase with rat skeletal muscle Na channels in Xenopus oocytes reduced the amplitude of Na currents and accelerated current decay. The effect required the presence of a potential phosphorylation site in the inactivation mechanism of the channel. The mutation responsible for human disease, trinucleotide repeats in the 3' untranslated region, did not prevent the effect. The consequence of an abnormal amount of the kinase would be altered muscle cell excitability, consistent with the clinical finding of myotonia in myotonic dystrophy. PMID:7738201

Mounsey, J P; Xu, P; John, J E; Horne, L T; Gilbert, J; Roses, A D; Moorman, J R

1995-05-01

167

Restoration of Urban River Water Quality: Introduction of Secondary Treated Domestic Wastewater Into the Nobidome Channel.  

National Technical Information Service (NTIS)

Comprehensive surveys for water quality and benthic invertebrates were carried out in the Nobidome Channel in which filtered secondary wastewater was, introduced to restore, the waterfront. The suspended solids concentration increased downstream as a resu...

M. Okada H. Kawahara S. Fukushima A. Mutoh

1992-01-01

168

Direct Interaction of Adenosine with the TRPV1 Channel Protein  

Microsoft Academic Search

Vanilloid receptor 1 (TRPV1), a nonspecific cation channel expressed primarily in small sensory neurons, mediates inflammatory thermal pain sensation. The function and expression of TRPV1 are enhanced during inflammation and certain neuropathies, leading to sustained hyperalgesia. Activation of TRPV1 in the spinal cord and periphery promotes release of adenosine, which produces analgesia by activating A1 and A2A adenosine receptor (AR)

Preeti Puntambekar; Jeremy Van Buren; Manish Raisinghani; Louis S. Premkumar; Vickram Ramkumar

2004-01-01

169

Ordinary High Water Mark in ephemeral and intermittent channels in the arid southwestern United States  

NASA Astrophysics Data System (ADS)

The Ordinary High Water Mark (OHWM) is important for regulating stream channels, as it defines the lateral extent of non-wetland waters, as defined under Waters of the United States in the Clean Water Act (33 CFR Part 328.3). Prior to this study, little was known about the OHWM and geomorphically effective events in arid southwest ephemeral and intermittent channels. The hydrogeomorphic floodplain features and flow dynamics are challenging to identify in these channels because of the unstable channel morphology, flashy flood events, and lack of gage data. Through field surveys, LiDAR high-resolution topography data collection, HEC-RAS flow modeling, analysis of aerial photography, and determination of recurrence intervals using gage data, we have developed a methodology to identify the hydrogeomorphic floodplain units and delineate the OHWM. The hydrogeomorphic floodplain units include a migratory low-flow channel, an active floodplain associated with low to moderate discharge events, and a low terrace inundated by moderate to high floods. We identified primary OHWM indicators such as a break in slope, a change in sediment texture, and a change in vegetation characteristics that create a repeatable and reliable signature in the channel morphology that is associated with the OHW boundary between the active floodplain and the low terrace. Secondary indicators such as drift, ripples, and silt deposits are randomly distributed throughout the channel and cannot be linked to a particular inundation extent. In an analysis of 14 gaged ephemeral and intermittent channels, we determined that the recurrence intervals varied from <1 to 15.5 yr. Lower recurrence intervals were typically associated with more stable channels, while channels with a sandy, erodable substrate had higher recurrence intervals. A more complete understanding of the channel morphology and the highly variable OHW recurrence intervals allows more reliable and repeatable delineations in southwest ephemeral and intermittent streams.

Curtis, K. E.; Lichvar, R.; Dixon, L.

2010-12-01

170

Cardiac sodium channel Na(v)1.5 and interacting proteins: Physiology and pathophysiology.  

PubMed

The cardiac voltage-gated Na(+) channel Na(v)1.5 generates the cardiac Na(+) current (INa). Mutations in SCN5A, the gene encoding Na(v)1.5, have been linked to many cardiac phenotypes, including the congenital and acquired long QT syndrome, Brugada syndrome, conduction slowing, sick sinus syndrome, atrial fibrillation, and dilated cardiomyopathy. The mutations in SCN5A define a sub-group of Na(v)1.5/SCN5A-related phenotypes among cardiac genetic channelopathies. Several research groups have proposed that Na(v)1.5 may be part of multi-protein complexes composed of Na(v)1.5-interacting proteins which regulate channel expression and function. The genes encoding these regulatory proteins have also been found to be mutated in patients with inherited forms of cardiac arrhythmias. The proteins that associate with Na(v)1.5 may be classified as (1) anchoring/adaptor proteins, (2) enzymes interacting with and modifying the channel, and (3) proteins modulating the biophysical properties of Na(v)1.5 upon binding. The aim of this article is to review these Na(v)1.5 partner proteins and to discuss how they may regulate the channel's biology and function. These recent investigations have revealed that the expression level, cellular localization, and activity of Na(v)1.5 are finely regulated by complex molecular and cellular mechanisms that we are only beginning to understand. PMID:19744495

Abriel, Hugues

2010-01-01

171

Emodin augments calcium activated chloride channel in colonic smooth muscle cells by Gi/Go protein.  

PubMed

Emodin is a natural anthraquinone in rhubarb. It has been identified as a prokinetic drug for gastrointestinal motility in Chinese traditional medicine. Emodin contracts smooth muscle by increasing the concentration of intracellular Ca(2+). In many smooth muscles, increasing intracellular Ca(2+) activates Ca(2+)-activated Cl(-) channels (ClCA). The study was aimed to investigate the effects of emodin on ClCA channels in colonic smooth muscle. 4 channel physiology signal acquire system was used to measure isometric contraction of smooth muscle strips. ClCA currents were recorded by EPC10 with perforated whole cell model. Emodin contracted strips and cells in colonic smooth muscle and augmented ClCA currents. Niflumic acid (NFA) and 4', 4'-diisothiostilbene-2, 2-disulfonic acid (DIDS) blocked the effects. Gi/Go protein inhibits protein kinase A (PKA) and protein kinase C (PKC), and PKA and PKC reduced ClCA currents. Pertussis toxin (PTX, a special inhibitor of Gi/Go protein), 8-bromoadenosine 38, 58-cyclic monophosphate (8-BrcAMP, a membrane-permeant protein kinase A activator) and Phorbol-12-myristate-13-acetate (PMA, a membrane-permeant protein kinase C activator) inhibited the effects on ClCA currents significantly. Our findings suggest that emodin augments ClCA channels to contract smooth muscle in colon, and the effect is induced mostly by enhancement of membrane Gi/Go protein signal transducer pathway. PMID:19409890

Xu, Long; Ting-Lou; Lv, Nonghua; Zhu, Xuan; Chen, Youxiang; Yang, Jing

2009-08-01

172

14-3-3 protein regulation of proton pumps and ion channels  

Microsoft Academic Search

In addition to their regulation of cytoplasmic enzymes, the 14-3-3 proteins are important regulators of membrane localised proteins. In particular, many of the cells' ion pumps and channels are either directly or indirectly modulated by 14-3-3 proteins. Binding of 14-3-3 can lead to the activation of pump activity as in the case of the plasma membrane H+-ATPase or inhibition as

Tom D. Bunney; Paul W. J. van den Wijngaard; Albertus H. de Boer

2002-01-01

173

Electron spin echo envelope modulation (ESEEM) reveals water and phosphate interactions with the KcsA potassium channel  

PubMed Central

Electron spin-echo envelope modulation (ESEEM) spectroscopy is a well-established technique for the study of naturally occurring paramagnetic metal centers. The technique has been used to study copper complexes, hemes, enzyme mechanisms, micellar water content, and water permeation profiles in membranes, among other applications. In the present study, we combine ESEEM spectroscopy with site-directed spin labeling (SDSL) and X-ray crystallography in order to evaluate the technique's potential as a structural tool to describe the native environment of membrane proteins. Using the KcsA potassium channel as a model system, we demonstrate that deuterium ESEEM can detect water permeation along the lipid-exposed surface of the KcsA outer helix. We further demonstrate that 31P ESEEM is able to identify channel residues that interact with the phosphate head group of the lipid bilayer. In combination with X-ray crystallography, the 31P data may be used to define the phosphate interaction surface of the protein. The results presented here establish ESEEM as a highly informative technique for SDSL studies of membrane proteins.

Cieslak, John A.; Focia, Pamela J.; Gross, Adrian

2010-01-01

174

Affinity of C60 neat fullerenes with membrane proteins: a computational study on potassium channels.  

PubMed

Most studies of the interactions of neat and functionalized fullerenes with cells have focused so far on their ability to cross the cell membrane envelopes. Membranes are, however, also host to a large number of proteins responsible for various cellular functions. Among these, ion channels are prominent components of the nervous system. Recently, it was shown that fullerenes may act as blockers or modulators of a variety of K+ channels. Here we use computer simulations to investigate the propensity of such nanocompounds to bind to K+ channels. Our results based on extensive atomistic molecular dynamics simulations reveal a variety of specific binding sites depending on the structure and properties of the channel. The corresponding binding free energies and putative mechanisms suggest that C60 may indeed effectively hinder the function of K+ channels and hence induce toxicity. PMID:20568711

Kraszewski, Sebastian; Tarek, Mounir; Treptow, Werner; Ramseyer, Christophe

2010-07-27

175

Dengue virus M protein C-terminal peptide (DVM-C) forms ion channels.  

PubMed

A chemically synthesized peptide consisting of the C-terminus of the M protein of the Dengue virus type 1 strain Singapore S275/90 (DVM-C) produced ion channel activity in artificial lipid bilayers. The channels had a variable conductance and were more permeable to sodium and potassium ions than to chloride ions and more permeable to chloride ions than to calcium ions. Hexamethylene amiloride (100 microM) and amantadine (10 microM), blocked channels formed by DVM-C. Ion channels may play an important role in the life cycle of many viruses and drugs that block these channels may prove to be useful antiviral agents. PMID:16007501

Premkumar, A; Horan, C R; Gage, P W

2005-03-01

176

Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines  

Microsoft Academic Search

BACKGROUND: Previous data from our laboratory has indicated that there is a functional link between the ?-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. METHODS: GIRK1-4 expression and levels

Howard K Plummer III; Madhu S Dhar; Maria Cekanova; Hildegard M Schuller

2005-01-01

177

Evaluation of Protein Reduction and Lysine Supplementation of Production Diets for Channel Catfish  

Microsoft Academic Search

A 2-year continuous production trial was conducted in earthen ponds to evaluate lysine supplementation of practical diets as a means of reducing the need for total dietary protein and limiting the nitrogenous waste production of channel catfish Ictalurus punctatus. Fingerling channel catfish with an initial weight of 34.1 g were stocked in twelve 0.04-ha ponds in early summer at a

T. Gibson Gaylord; Wendy M. Sealey; Delbert M. Gatlin III

2002-01-01

178

G protein control of potassium channel activity in a mast cell line  

Microsoft Academic Search

Using the patch-clamp technique, we studied regulation of potassium channels by G protein activators in the histamine-secreting rat basophilic leukemia (RBL-2H3) cell line. These cells normally express inward rectifier K + channels, with a macroscopic whole-cell conductance in normal Ringer ranging from 1 to 16 nS\\/cell. This conductance is stabilized by including ATP or GTP in the pipette solution. Intracellular

M. A. McCloskey; MICHAEL D. CAHALAN

1990-01-01

179

Mechanisms underlying the protein-kinase mediated regulation of the HERG potassium channel synthesis  

PubMed Central

The HERG (human ether-a-go-go related gene) potassium channel aids in repolarization of the cardiomyocyte membrane at the end of each action potential. We have previously shown that sustained protein kinase A or C (PKA and PKC) activity specifically enhances channel synthesis over the course of hours to days in heterologous expression and cardiac myocytes. The kinase-mediated augmentation of the channel is post-transcriptional and occurs near or at the endoplasmic reticulum. Here we report our further investigations into the mechanisms of kinase-mediated augmentation of HERG channel protein. We show that HERG channel phosphorylation alone is not sufficient for the PKA-dependent increase to occur. In vitro translation studies indicate that an additional factor is required for the process. Pharmacologic inhibitors suggest that the channel augmentation is not due to kinase-mediated alteration in proteasome or lysosome activity. PKA activation had no effect on stability of HERG mRNA and polyribosomal profiling showed that kinase activity did not elevate translation from low to high rates. Transcriptional inhibition results suggest that the additional cellular factor is a PKA-regulated protein. Together, these findings suggest that PKA-mediated augmentation of HERG abundance is more complex than previously appreciated involving enhancement of already active translation rates, phosphorylation of the channel protein and at least one other cAMP/PKA-responsive protein. Further exploration of molecular components of this regulatory pathway will be necessary to determine exact mechanism and the biomedical impact of this process in vivo.

Krishnan, Yamini; Li, Yan; Zheng, Renjian; Kanda, Vikram; McDonald, Thomas V.

2012-01-01

180

Membrane Incorporation, Channel Formation, and Disruption of Calcium Homeostasis by Alzheimer's ?-Amyloid Protein  

PubMed Central

Oligomerization, conformational changes, and the consequent neurodegeneration of Alzheimer's ?-amyloid protein (A?P) play crucial roles in the pathogenesis of Alzheimer's disease (AD). Mounting evidence suggests that oligomeric A?Ps cause the disruption of calcium homeostasis, eventually leading to neuronal death. We have demonstrated that oligomeric A?Ps directly incorporate into neuronal membranes, form cation-sensitive ion channels (“amyloid channels”), and cause the disruption of calcium homeostasis via the amyloid channels. Other disease-related amyloidogenic proteins, such as prion protein in prion diseases or ?-synuclein in dementia with Lewy bodies, exhibit similarities in the incorporation into membranes and the formation of calcium-permeable channels. Here, based on our experimental results and those of numerous other studies, we review the current understanding of the direct binding of A?P into membrane surfaces and the formation of calcium-permeable channels. The implication of composition of membrane lipids and the possible development of new drugs by influencing membrane properties and attenuating amyloid channels for the treatment and prevention of AD is also discussed.

Kawahara, Masahiro; Ohtsuka, Isao; Yokoyama, Shoko; Kato-Negishi, Midori; Sadakane, Yutaka

2011-01-01

181

Pxmp2 Is a Channel-Forming Protein in Mammalian Peroxisomal Membrane  

PubMed Central

Background Peroxisomal metabolic machinery requires a continuous flow of organic and inorganic solutes across peroxisomal membrane. Concerning small solutes, the molecular nature of their traffic has remained an enigma. Methods/Principal Findings In this study, we show that disruption in mice of the Pxmp2 gene encoding Pxmp2, which belongs to a family of integral membrane proteins with unknown function, leads to partial restriction of peroxisomal membrane permeability to solutes in vitro and in vivo. Multiple-channel recording of liver peroxisomal preparations reveals that the channel-forming components with a conductance of 1.3 nS in 1.0 M KCl were lost in Pxmp2?/? mice. The channel-forming properties of Pxmp2 were confirmed with recombinant protein expressed in insect cells and with native Pxmp2 purified from mouse liver. The Pxmp2 channel, with an estimated diameter of 1.4 nm, shows weak cation selectivity and no voltage dependence. The long-lasting open states of the channel indicate its functional role as a protein forming a general diffusion pore in the membrane. Conclusions/Significance Pxmp2 is the first peroxisomal channel identified, and its existence leads to prediction that the mammalian peroxisomal membrane is permeable to small solutes while transfer of “bulky” metabolites, e.g., cofactors (NAD/H, NADP/H, and CoA) and ATP, requires specific transporters.

Rokka, Aare; Soininen, Raija; Immonen, Hanna L.; Pirila, Paivi L.; Bergmann, Ulrich; Sormunen, Raija T.; Weckstrom, Matti; Hiltunen, J. Kalervo

2009-01-01

182

The study of venous channel water pollution load reduction technologies  

Microsoft Academic Search

In this paper, Hongqi River of WuJin River watershed (china) was selected as an example, aims to use QUAL2K water quality model simulation water environmental capacity of the Hongqi River, forecasting the pollution load of the Hongqi River basin, and compared to the actual emissions of pollution load, calculated the load reduction rate for satisfying the water quality objectives. At

Ruibin Zhang; Yanyan Dou; Ping Sun; Yiyao Bai; Xin Qian

2011-01-01

183

Aquaporin3 is a sperm water channel essential for postcopulatory sperm osmoadaptation and migration.  

PubMed

In the journey from the male to female reproductive tract, mammalian sperm experience a natural osmotic decrease (e.g., in mouse, from ~415 mOsm in the cauda epididymis to ~310 mOsm in the uterine cavity). Sperm have evolved to utilize this hypotonic exposure for motility activation, meanwhile efficiently silence the negative impact of hypotonic cell swelling. Previous physiological and pharmacological studies have shown that ion channel-controlled water influx/efflux is actively involved in the process of sperm volume regulation; however, no specific sperm proteins have been found responsible for this rapid osmoadaptation. Here, we report that aquaporin3 (AQP3) is a sperm water channel in mice and humans. Aqp3-deficient sperm show normal motility activation in response to hypotonicity but display increased vulnerability to hypotonic cell swelling, characterized by increased tail bending after entering uterus. The sperm defect is a result of impaired sperm volume regulation and progressive cell swelling in response to physiological hypotonic stress during male-female reproductive tract transition. Time-lapse imaging revealed that the cell volume expansion begins at cytoplasmic droplet, forcing the tail to angulate and form a hairpin-like structure due to mechanical membrane stretch. The tail deformation hampered sperm migration into oviduct, resulting in impaired fertilization and reduced male fertility. These data suggest AQP3 as an essential membrane pathway for sperm regulatory volume decrease (RVD) that balances the "trade-off" between sperm motility and cell swelling upon physiological hypotonicity, thereby optimizing postcopulatory sperm behavior. PMID:21135872

Chen, Qi; Peng, Hongying; Lei, Li; Zhang, Ying; Kuang, Haibin; Cao, Yujing; Shi, Qi-Xian; Ma, Tonghui; Duan, Enkui

2011-06-01

184

Identification and characterisation of a functional aquaporin water channel (Anomala cuprea DRIP) in a coleopteran insect.  

PubMed

Water transport across the plasma membrane depends on the presence of the water channel aquaporin (AQP), which mediates the bulk movement of water through osmotic and pressure gradients. In terrestrial insects, which are solid and/or plant feeders, the entrance and exit of water is primarily executed along the alimentary tract, where the hindgut, particularly the rectum, is the major site of water conservation. A cDNA encoding the homologue of the water-specific Drosophila AQP [Drosophila integral protein (DRIP)] was identified through the RT-PCR of RNA isolated from the rectum of the cupreous chafer larvae, Anomala cuprea, a humus and plant root feeder. This gene (Anocu AQP1) has a predicted molecular mass of 26.471 kDa, similar to the DRIP clade of insect AQPs characterised from caterpillars, flies and several liquid-feeding insects. When expressed in Xenopus laevis oocytes, Anocu AQP1 showed the hallmarks of aquaporin-mediated water transport but no glycerol or urea permeability, and the reversible inhibition of elevated water transport through 1 mmol l(-1) HgCl2. This is the first experimental demonstration of the presence of a water-specific AQP, namely DRIP, in the Coleoptera. The genome of the model beetle Tribolium castaneum contains six putative AQP sequences, one of which (Trica-1a, XP_972862) showed the highest similarity to Anocu AQP1 (~60% amino acid identity). Anocu AQP1 is predominantly expressed in the rectum. Using a specific antibody raised against DRIP in the silkworm Bombyx mori (AQP-Bom1), Anocu AQP1 was localised to the apical plasma membrane of rectal epithelial cells, and lacking in the midgut and gastric caecal epithelia. Based on the BeetleBase prediction, there are three putative AQPs (Trica-3a, 3b, 3c: XP_970728, 970912, 970791) that are homologous to B. mori aquaglyceroporin [AQP-Bom2 (GLP)]. The immunocytochemical studies using the specific anti-peptide antibody against AQP-Bom2 revealed the presence of the GLP homologue at the apical plasma membrane of enterocytes in the midgut and gastric caeca. Thus, DRIP (Anocu AQP1) and the putative GLP share epithelial fluid-transporting roles along the alimentary tract in cupreous chafer larvae. PMID:23531819

Nagae, Tomone; Miyake, Seiji; Kosaki, Shiho; Azuma, Masaaki

2013-07-15

185

Mercury-sensitive water channels as possible sensors of water potentials in pollen  

PubMed Central

The growing pollen tube is central to plant reproduction and is a long-standing model for cellular tip growth in biology. Rapid osmotically driven growth is maintained under variable conditions, which requires osmosensing and regulation. This study explores the mechanism of water entry and the potential role of osmosensory regulation in maintaining pollen growth. The osmotic permeability of the plasmalemma of Lilium pollen tubes was measured from plasmolysis rates to be 1.32±0.31×10–3 cm s–1. Mercuric ions reduce this permeability by 65%. Simulations using an osmotic model of pollen tube growth predict that an osmosensor at the cell membrane controls pectin deposition at the cell tip; inhibiting the sensor is predicted to cause tip bursting due to cell wall thinning. It was found that adding mercury to growing pollen tubes caused such a bursting of the tips. The model indicates that lowering the osmotic permeability per se does not lead to bursting but rather to thickening of the tip. The time course of induced bursting showed no time lag and was independent of mercury concentration, compatible with a surface site of action. The submaximal bursting response to intermediate mercuric ion concentration was independent of the concentration of calcium ions, showing that bursting is not due to a competitive inhibition of calcium binding or entry. Bursting with the same time course was also shown by cells growing on potassium-free media, indicating that potassium channels (implicated in mechanosensing) are not involved in the bursting response. The possible involvement of mercury-sensitive water channels as osmosensors and current knowledge of these in pollen cells are discussed.

Hill, Adrian E.

2013-01-01

186

Mercury-sensitive water channels as possible sensors of water potentials in pollen.  

PubMed

The growing pollen tube is central to plant reproduction and is a long-standing model for cellular tip growth in biology. Rapid osmotically driven growth is maintained under variable conditions, which requires osmosensing and regulation. This study explores the mechanism of water entry and the potential role of osmosensory regulation in maintaining pollen growth. The osmotic permeability of the plasmalemma of Lilium pollen tubes was measured from plasmolysis rates to be 1.32±0.31×10(-3) cm s(-1). Mercuric ions reduce this permeability by 65%. Simulations using an osmotic model of pollen tube growth predict that an osmosensor at the cell membrane controls pectin deposition at the cell tip; inhibiting the sensor is predicted to cause tip bursting due to cell wall thinning. It was found that adding mercury to growing pollen tubes caused such a bursting of the tips. The model indicates that lowering the osmotic permeability per se does not lead to bursting but rather to thickening of the tip. The time course of induced bursting showed no time lag and was independent of mercury concentration, compatible with a surface site of action. The submaximal bursting response to intermediate mercuric ion concentration was independent of the concentration of calcium ions, showing that bursting is not due to a competitive inhibition of calcium binding or entry. Bursting with the same time course was also shown by cells growing on potassium-free media, indicating that potassium channels (implicated in mechanosensing) are not involved in the bursting response. The possible involvement of mercury-sensitive water channels as osmosensors and current knowledge of these in pollen cells are discussed. PMID:24098048

Shachar-Hill, Bruria; Hill, Adrian E; Powell, Janet; Skepper, Jeremy N; Shachar-Hill, Yair

2013-11-01

187

Voltage-gated sodium channel (NaV) protein dissection creates a set of functional pore-only proteins.  

PubMed

Many voltage-gated ion channel (VGIC) superfamily members contain six-transmembrane segments in which the first four form a voltage-sensing domain (VSD) and the last two form the pore domain (PD). Studies of potassium channels from the VGIC superfamily together with identification of voltage-sensor only proteins have suggested that the VSD and the PD can fold independently. Whether such transmembrane modularity is common to other VGIC superfamily members has remained untested. Here we show, using protein dissection, that the Silicibacter pomeroyi voltage-gated sodium channel (Na(V)Sp1) PD forms a stand-alone, ion selective pore (Na(V)Sp1p) that is tetrameric, ?-helical, and that forms functional, sodium-selective channels when reconstituted into lipid bilayers. Mutation of the Na(V)Sp1p selectivity filter from LESWSM to LDDWSD, a change similar to that previously shown to alter ion selectivity of the bacterial sodium channel Na(V)Bh1 (NaChBac), creates a calcium-selective pore-only channel, Ca(V)Sp1p. We further show that production of PDs can be generalized by making pore-only proteins from two other extremophile Na(V)s: one from the hydrocarbon degrader Alcanivorax borkumensis (Na(V)Ab1p), and one from the arsenite oxidizer Alkalilimnicola ehrlichei (Na(V)Ae1p). Together, our data establish a family of active pore-only ion channels that should be excellent model systems for study of the factors that govern both sodium and calcium selectivity and permeability. Further, our findings suggest that similar dissection approaches may be applicable to a wide range of VGICs and, thus, serve as a means to simplify and accelerate biophysical, structural, and drug development efforts. PMID:21746903

Shaya, David; Kreir, Mohamed; Robbins, Rebecca A; Wong, Stephanie; Hammon, Justus; Brüggemann, Andrea; Minor, Daniel L

2011-07-26

188

Structure, dynamics and reactions of protein hydration water.  

PubMed Central

The apparent simplicity of the water molecule belies the wide range of fascinating protein phenomena in which it participates. We review recent computer simulation work on buried, internal water molecules, discussing the thermodynamics of water molecule binding and the participation of water in proton transfer reactions. Surface water molecules are also considered, with emphasis on the modification of average solvent structure on a protein surface, the role of water in the protein dynamical 'glass' transition and a simplified description of the protein motions thereby activated.

Smith, Jeremy C; Merzel, Franci; Bondar, Ana-Nicoleta; Tournier, Alexander; Fischer, Stefan

2004-01-01

189

Pore waters regulate ion permeation in a calcium release-activated calcium channel  

PubMed Central

The recent crystal structure of Orai, the pore unit of a calcium release-activated calcium (CRAC) channel, is used as the starting point for molecular dynamics and free-energy calculations designed to probe this channel’s conduction properties. In free molecular dynamics simulations, cations localize preferentially at the extracellular channel entrance near the ring of Glu residues identified in the crystal structure, whereas anions localize in the basic intracellular half of the pore. To begin to understand ion permeation, the potential of mean force (PMF) was calculated for displacing a single Na+ ion along the pore of the CRAC channel. The computed PMF indicates that the central hydrophobic region provides the major hindrance for ion diffusion along the permeation pathway, thereby illustrating the nonconducting nature of the crystal structure conformation. Strikingly, further PMF calculations demonstrate that the mutation V174A decreases the free energy barrier for conduction, rendering the channel effectively open. This seemingly dramatic effect of mutating a nonpolar residue for a smaller nonpolar residue in the pore hydrophobic region suggests an important role for the latter in conduction. Indeed, our computations show that even without significant channel-gating motions, a subtle change in the number of pore waters is sufficient to reshape the local electrostatic field and modulate the energetics of conduction, a result that rationalizes recent experimental findings. The present work suggests the activation mechanism for the wild-type CRAC channel is likely regulated by the number of pore waters and hence pore hydration governs the conductance.

Dong, Hao; Fiorin, Giacomo; Carnevale, Vincenzo; Treptow, Werner; Klein, Michael L.

2013-01-01

190

OPTIMIZATION OF WATER CHANNEL DESIGN IN BEAM-BLANK MOLDS  

Microsoft Academic Search

The magnitude and uniformity of the hotface temperature greatly affects the life of continuous casting molds, in addition to affecting surface defects in the product, such as longitudinal cracks. In this work, several different designs for the cooling channels of a beam-blank mold are evaluated using two-dimensional finite-element heat conduction models in order to minimize hotface temperature and to improve

B. G. Thomas; J. Jiang; D. Lorento

191

Asymmetric fluxes of water and sediments in a mesotidal mudflat channel  

NASA Astrophysics Data System (ADS)

The hydrodynamics of a small tributary channel and its adjacent mudflat is studied in Willapa Bay, Washington State, USA. Velocity profiles and water levels are simultaneously measured at different locations in the channel and on the mudflat for two weeks. The above tidal flat and channel hydrodynamics differ remarkably during the tidal cycle. When the water surface level is above the tidal flat elevation, the channel is inactive. At this stage, the above tidal flat flow is predominantly aligned along the Bay axis, oscillating with the tide as a standing wave with peak velocities up to 0.3 m/s. When the mudflat becomes emergent, the flow concentrates in the channel. During this stage, current velocities up to 1 m/s are measured during ebb; and up to 0.6 m/s during flood. Standard equations for open-channel flow are utilized to study the channel hydrodynamics. From the continuity equation, a lateral inflow is predicted during ebb, which likely originates from the drainage of the mudflat through the lateral runnels. Both advective acceleration and lateral discharge terms, estimated directly from the velocity profiles, play a significant role in the momentum equation. The computed drag coefficient for bottom friction is small, due to an absence of vegetation and bottom bedforms in the channel. Sediment fluxes are calculated by combining flow and suspended sediment concentration estimated using the acoustic backscatter signal of the instruments. A net export of the sediment from the channel is found during ebb, which is not balanced by the sediment import during flood. When the mudflat is submerged, ebb-flood asymmetries in suspended sediment concentration are present, leading to a net sediment flux toward the inner part of the Willapa Bay. Finally, a residual flow is detected inside the channel at high slack water, probably associated with the thermohaline circulation.

Mariotti, G.; Fagherazzi, S.

2011-01-01

192

Water and sediment transport of channel-flat systems in a mesotidal mudflat: Willapa Bay, Washington  

NASA Astrophysics Data System (ADS)

The muddy tidal flats of southern Willapa Bay, Washington are tidally dominated and receive little direct freshwater input. We use data from instruments deployed in channels of different size and on their adjacent flats to investigate the hydrodynamics and sediment dynamics of each morphological setting under a range of seasonal and meteorological conditions, including rain and wind events. Interaction between the morphology of the channel/flat complex and tidal water-level variations produces well-defined velocity pulses during both flooding and ebbing tides. These pulses represent about 27% of the total along-channel water transport and 35% of the suspended-sediment transport of the system. Maintenance of continuity produces the velocity pulse, and pulse magnitude is determined by tidal range. Wind alters the flow regime in channels and on the flat, enhancing over-flat ebb flow in this study location while decreasing ebb-pulse intensity. Wind speed was positively correlated with minimum suspended-sediment concentration. Precipitation falling directly on flats was found to erode flat sediment, which subsequently formed a temporary deposit in the adjacent channel. Residual along-channel water transport in channels and on nearby flats was flood dominant under all seasonal conditions sampled, and sediment flux was flood dominant during winter and spring deployments.

Nowacki, Daniel J.; Ogston, Andrea S.

193

Water and sediment transport of channel-flat systems in a mesotidal mudflat: Willapa Bay, Washington  

NASA Astrophysics Data System (ADS)

The muddy tidal flats of southern Willapa Bay, Washington are tidally dominated and receive little direct freshwater input. We use data from instruments deployed in channels of different size and on their adjacent flats to investigate the hydrodynamics and sediment dynamics of each morphological setting under a range of seasonal and meteorological conditions, including rain and wind events. Interaction between the morphology of the channel/flat complex and tidal water-level variations produces well-defined velocity pulses during both flooding and ebbing tides. These pulses represent about 27% of the total along-channel water transport and 35% of the suspended-sediment transport of the system. Maintenance of continuity produces the velocity pulse, and pulse magnitude is determined by tidal range. Wind alters the flow regime in channels and on the flat, enhancing over-flat ebb flow in this study location while decreasing ebb-pulse intensity. Wind speed was positively correlated with minimum suspended-sediment concentration. Precipitation falling directly on flats was found to erode flat sediment, which subsequently formed a temporary deposit in the adjacent channel. Residual along-channel water transport in channels and on nearby flats was flood dominant under all seasonal conditions sampled, and sediment flux was flood dominant during winter and spring deployments.

Nowacki, Daniel J.; Ogston, Andrea S.

2013-06-01

194

Behavior of Interfacial Waves in Cocurrent Oil-Water Channel Flow  

Microsoft Academic Search

Measurements of the spectral behavior of interfacial waves in cocurrent oil-water pressure driven channel flow are obtained and compared to a weakly nonlinear simulation. The goal is to develop an understanding of the utility and limitations of linear and weakly nonliner theories in two layer flows. Experiments are conducted with water and a light hydrocarbon oil with a density of

Bennett Woods; Mark McCready

1999-01-01

195

An improved vertical-axis water-current turbine incorporating a channelling device  

Microsoft Academic Search

Water-Current Turbines (WCTs) are non-polluting electricity generation plants that harness the kinetic energy of natural water courses, using several kinds of rotors. At the School of Engineering of the University of Buenos Aires, researchers are developing a WCT whose particular characteristics improve technical and economic performance. A channelling device, integrated into the flotation system, is used to modify flow conditions

Fernando Ponta; Gautam Shankar Dutt

2000-01-01

196

Cooperative endocytosis of the endosomal SNARE protein syntaxin-8 and the potassium channel TASK-1.  

PubMed

The endosomal SNARE protein syntaxin-8 interacts with the acid-sensitive potassium channel TASK-1. The functional relevance of this interaction was studied by heterologous expression of these proteins (and mutants thereof) in Xenopus oocytes and in mammalian cell lines. Coexpression of syntaxin-8 caused a fourfold reduction in TASK-1 current, a corresponding reduction in the expression of TASK-1 at the cell surface, and a marked increase in the rate of endocytosis of the channel. TASK-1 and syntaxin-8 colocalized in the early endosomal compartment, as indicated by the endosomal markers 2xFYVE and rab5. The stimulatory effect of the SNARE protein on the endocytosis of the channel was abolished when both an endocytosis signal in TASK-1 and an endocytosis signal in syntaxin-8 were mutated. A syntaxin-8 mutant that cannot assemble with other SNARE proteins had virtually the same effect as wild-type syntaxin-8. Total internal reflection fluorescence microscopy showed formation and endocytosis of vesicles containing fluorescence-tagged clathrin, TASK-1, and/or syntaxin-8. Our results suggest that the unassembled form of syntaxin-8 and the potassium channel TASK-1 are internalized via clathrin-mediated endocytosis in a cooperative manner. This implies that syntaxin-8 regulates the endocytosis of TASK-1. Our study supports the idea that endosomal SNARE proteins can have functions unrelated to membrane fusion. PMID:24743596

Renigunta, Vijay; Fischer, Thomas; Zuzarte, Marylou; Kling, Stefan; Zou, Xinle; Siebert, Kai; Limberg, Maren M; Rinné, Susanne; Decher, Niels; Schlichthörl, Günter; Daut, Jürgen

2014-06-15

197

The threshold of vapor channel formation in water induced by pulsed CO2 laser  

NASA Astrophysics Data System (ADS)

Water plays an important role in laser ablation. There are two main interpretations of laser-water interaction: hydrokinetic effect and vapor phenomenon. The two explanations are reasonable in some way, but they can't explain the mechanism of laser-water interaction completely. In this study, the dynamic process of vapor channel formation induced by pulsed CO2 laser in static water layer was monitored by high-speed camera. The wavelength of pulsed CO2 laser is 10.64 um, and pulse repetition rate is 60 Hz. The laser power ranged from 1 to 7 W with a step of 0.5 W. The frame rate of high-speed camera used in the experiment was 80025 fps. Based on high-speed camera pictures, the dynamic process of vapor channel formation was examined, and the threshold of vapor channel formation, pulsation period, the volume, the maximum depth and corresponding width of vapor channel were determined. The results showed that the threshold of vapor channel formation was about 2.5 W. Moreover, pulsation period, the maximum depth and corresponding width of vapor channel increased with the increasing of the laser power.

Guo, Wenqing; Zhang, Xianzeng; Zhan, Zhenlin; Xie, Shusen

2012-12-01

198

The Small Hydrophobic Protein of the Human Respiratory Syncytial Virus Forms Pentameric Ion Channels*  

PubMed Central

The small hydrophobic (SH) protein is encoded by the human respiratory syncytial virus. Its absence leads to viral attenuation in the context of whole organisms, and it prevents apoptosis in infected cells. Herein, we have examined the structure of SH protein in detergent micelles and in lipid bilayers, by solution NMR and attenuated total reflection-Fourier transform infrared spectroscopy, respectively. We found that SH protein has a single ?-helical transmembrane domain and forms homopentamers in several detergents. In detergent micelles, the transmembrane domain is flanked N-terminally by an ?-helix that forms a ring around the lumen of the pore and C-terminally by an extended ?-turn. SH protein was found in the plasma membrane of transiently expressing HEK 293 cells, which showed pH-dependent (acid-activated) channel activity. Channel activity was abolished in mutants lacking both native His residues, His22 and His51, but not when either His was present. Herein, we propose that the pentameric model of SH protein presented is a physiologically relevant conformation, albeit probably not the only one, in which SH contributes to RSV infection and replication. Viroporins are short (?100 amino acids) viral membrane proteins that form oligomers of a defined size, act as proton or ion channels, and in general enhance membrane permeability in the host. However, with some exceptions, their precise biological role of their channel activity is not understood. In general, viroporins resemble poorly specialized proteins but are nevertheless critical for viral fitness. In vivo, viruses lacking viroporins usually exhibit an attenuated or weakened phenotype, altered tropism, and diminished pathological effects. We have chosen to study the SH protein, 64 amino acids long, found in the human respiratory syncytial virus because of the effect of RSV on human health and the lack of adequate antivirals. We show that SH protein forms oligomers that behave as ion channels when activated at low pH. This study adds SH protein to a growing group of viroporins that have been structurally characterized. Although the precise biological role of this pentameric channel is still unknown, this report is nevertheless essential to fill some of the many gaps that exist in the understanding of SH protein function.

Gan, Siok-Wan; Tan, Edward; Lin, Xin; Yu, Dejie; Wang, Juejin; Tan, Gregory Ming-Yeong; Vararattanavech, Ardcharaporn; Yeo, Chiew Ying; Soon, Cin Huang; Soong, Tuck Wah; Pervushin, Konstantin; Torres, Jaume

2012-01-01

199

Effect of heavy water on protein flexibility.  

PubMed

The effects of heavy water (D(2)O) on internal dynamics of proteins were assessed by both the intrinsic phosphorescence lifetime of deeply buried Trp residues, which reports on the local structure about the triplet probe, and the bimolecular acrylamide phosphorescence quenching rate constant that is a measure of the average acrylamide diffusion coefficient through the macromolecule. The results obtained with several protein systems (ribonuclease T1, superoxide dismutase, beta-lactoglobulin, liver alcohol dehydrogenase, alkaline phosphatase, and apo- and Cd-azurin) demonstrate that in most cases D(2)O does significantly increase the rigidity the native structure. With the exception of alkaline phosphatase, the kinetics of the structure tightening effect of deuteration are rapid compared with the rate of H/D exchange of internal protons, which would then assign the dampening of structural fluctuations in D(2)O to a solvent effect, rather than to stronger intramolecular D bonding. Structure tightening by heavy water is generally amplified at higher temperatures, supporting a mostly hydrophobic nature of the underlying interaction, and under conditions that destabilize the globular fold. PMID:12023248

Cioni, Patrizia; Strambini, Giovanni B

2002-06-01

200

Interfacial complexes between a protein and lipophilic ions at an oil-water interface.  

PubMed

The interaction between an intact protein and two lipophilic ions at an oil-water interface has been investigated using cyclic voltammetry, impedance based techniques and a newly developed method in which the biphasic oil-water system is analyzed by biphasic electrospray ionization mass spectrometry (BESI-MS), using a dual-channel electrospray emitter. It is found that the protein forms interfacial complexes with the lipophilic ions and that it specifically requires the presence of the oil-water interface to be formed under the experimental conditions. Furthermore, impedance based techniques and BESI-MS with a common ion to polarize the interface indicated that the Galvani potential difference across the oil-water interface significantly influences the interfacial complexation degree. The ability to investigate protein-ligand complexes formed at polarized liquid-liquid interfaces is thus a new analytical method for assessing potential dependent interfacial complexation using a structure elucidating detection principle. PMID:20735009

Hartvig, Rune A; Méndez, Manuel A; van de Weert, Marco; Jorgensen, Lene; Østergaard, Jesper; Girault, Hubert H; Jensen, Henrik

2010-09-15

201

p.R254Q mutation in the aquaporin-2 water channel causing dominant nephrogenic diabetes insipidus is due to a lack of arginine vasopressin-induced phosphorylation  

Microsoft Academic Search

Vasopressin regulates human water homeostasis by re-distributing homotetrameric aquaporin-2 (AQP2) water channels from intracellular vesicles to the apical membrane of renal principal cells, a process in which phosphorylation of AQP2 at S256 by cAMP-dependent protein kinase A (PKA) is thought to be essential. Dominant nephrogenic diabetes insipidus (NDI), a disease in which the kidney is unable to concentrate urine in

Paul JM Savelkoul; Fabrizio De Mattia; Yuedan Li; Erik-Jan Kamsteeg; Irene BM Konings; Peter van der Sluijs; Peter MT Deen

2009-01-01

202

33 CFR 207.640 - Sacramento Deep Water Ship Channel Barge Lock and Approach Canals; use, administration, and...  

Code of Federal Regulations, 2013 CFR

33 Navigation and Navigable Waters 3 2013-07-01 2013-07-01 false Sacramento Deep Water Ship Channel Barge Lock and Approach Canals...Section 207.640 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF...

2013-07-01

203

33 CFR 207.640 - Sacramento Deep Water Ship Channel Barge Lock and Approach Canals; use, administration, and...  

Code of Federal Regulations, 2011 CFR

33 Navigation and Navigable Waters 3 2011-07-01 2011-07-01 false Sacramento Deep Water Ship Channel Barge Lock and Approach Canals...Section 207.640 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF...

2011-07-01

204

33 CFR 207.640 - Sacramento Deep Water Ship Channel Barge Lock and Approach Canals; use, administration, and...  

Code of Federal Regulations, 2012 CFR

33 Navigation and Navigable Waters 3 2012-07-01 2012-07-01 false Sacramento Deep Water Ship Channel Barge Lock and Approach Canals...Section 207.640 Navigation and Navigable Waters CORPS OF ENGINEERS, DEPARTMENT OF...

2012-07-01

205

Gating and conduction of nano-channel forming proteins: a computational approach.  

PubMed

Monitoring conformational changes in ion channels is essential to understand their gating mechanism. Here, we explore the structural dynamics of four outer membrane proteins with different structures and functions in the slowest nonzero modes of vibration. Normal mode analysis was performed on the modified elastic network model of channel in the membrane. According to our results, when membrane proteins were analyzed in the dominant mode, the composed pores, TolC and ?-hemolysin showed large motions at the intramembrane ?-barrel region while, in other porins, OmpA and OmpF, largest motions observed in the region of external flexible loops. A criterion based on equipartition theorem was used to measure the possible amplitude of vibration in channel forming proteins. The current approach complements theoretical and experimental techniques including HOLE, Molecular Dynamics (MD), and voltage clamp used to address the channel's structure and dynamics and provides the means to conduct a theoretical simultaneous study of the structure and function of the channel. An animated interactive 3D complement (I3DC) is available in Proteopedia at http://proteopedia.org/w/Journal:JBSD:3. PMID:22928968

Besya, A B; Mobasheri, H; Ejtehadi, M R

2013-01-01

206

IgG marker of optic-spinal multiple sclerosis binds to the aquaporin-4 water channel  

PubMed Central

Neuromyelitis optica (NMO) is an inflammatory demyelinating disease that selectively affects optic nerves and spinal cord. It is considered a severe variant of multiple sclerosis (MS), and frequently is misdiagnosed as MS, but prognosis and optimal treatments differ. A serum immunoglobulin G autoantibody (NMO-IgG) serves as a specific marker for NMO. Here we show that NMO-IgG binds selectively to the aquaporin-4 water channel, a component of the dystroglycan protein complex located in astrocytic foot processes at the blood-brain barrier. NMO may represent the first example of a novel class of autoimmune channelopathy.

Lennon, Vanda A.; Kryzer, Thomas J.; Pittock, Sean J.; Verkman, A.S.; Hinson, Shannon R.

2005-01-01

207

Preparation of semi-solid aluminum alloy slurry poured through a water-cooled serpentine channel  

NASA Astrophysics Data System (ADS)

A water-cooled serpentine channel pouring process was invented to produce semi-solid A356 aluminum alloy slurry for rheocasting, and the effects of pouring temperature and circulating cooling water flux on the microstructure of the slurry were investigated. The results show that at the pouring temperature of 640-680°C and the circulating cooling water flux of 0.9 m3/h, the semi-solid A356 aluminum alloy slurry with spherical primary ?(Al) grains can be obtained, whose shape factors are between 0.78 and 0.86 and the grain diameter can reach 48-68 ?m. When the pouring temperatures are at 660-680°C, only a very thin solidified shell remains inside the serpentine channel and can be removed easily. When the serpentine channel is cooled with circulating water, the microstructure of the semi-solid slurry can be improved, and the serpentine channel is quickly cooled to room temperature after the completion of one pouring. In terms of the productivity of the special equipment, the water-cooled serpentine channel is economical and efficient.

Chen, Zheng-Zhou; Mao, Wei-Min; Wu, Zong-Chuang

2012-01-01

208

The human AQP4 gene: definition of the locus encoding two water channel polypeptides in brain.  

PubMed Central

The aquaporin family of membrane water transport proteins are expressed in diverse tissues, and in brain the predominant water channel protein is AQP4. Here we report the isolation and characterization of the human AQP4 cDNAs and genomic DNA. Two cDNAs were isolated corresponding to the two initiating methionines (M1 in a 323-aa polypeptide and M23 in a 301-aa polypeptide) previously identified in rat [Jung, J.S., Bhat, R.V., Preston, G.M., Guggino, W.B. & Agre, P. (1994) Proc. Natl. Acad. Sci. USA 91, 13052-13056]. Similar to other aquaporins, the AQP4 gene is composed of four exons encoding 127, 55, 27, and 92 amino acids separated by introns of 0.8, 0.3, and 5.2 kb. Unlike other aquaporins, an alternative coding initiation sequence (designated exon 0) was located 2.7 kb upstream of exon 1. When spliced together, M1 and the subsequent 10 amino acids are encoded by exon 0; the next 11 amino acids and M23 are encoded by exon 1. Transcription initiation sites have been mapped in the proximal promoters of exons 0 and 1. RNase protection revealed distinct transcripts corresponding to M1 and M23 mRNAs, and AQP4 immunoblots of cerebellum demonstrated reactive polypeptides of 31 and 34 kDa. Using a P1 and a lambda EMBL subclone, the chromosomal site of the human AQP4 gene was mapped to chromosome 18 at the junction of q11.2 and q12.1 by fluorescence in situ hybridization. These studies may now permit molecular characterization of AQP4 during human development and in clinical disorders. Images Fig. 4 Fig. 5

Lu, M; Lee, M D; Smith, B L; Jung, J S; Agre, P; Verdijk, M A; Merkx, G; Rijss, J P; Deen, P M

1996-01-01

209

Protein methylation activates reconstituted ryanodine receptor-ca release channels from coronary artery myocytes.  

PubMed

Ryanodine receptors (RyR) play an important role in the regulation of intracellular Ca(2+) concentration and in the control of vascular tone. However, the mechanism regulating the activity of RyR is poorly understood. The present study determined whether protein methylation participates in the control of RyR activity. Using a planar lipid bilayer clamping system, S-adenosyl-L-methionine (SAM), a methyl donor, significantly increased the activity of a 245-pS reconstituted Ca(2+) release channel from coronary arterial smooth muscle (CASM) in a concentration-dependent manner. Addition of the protein methylation blockers, 3-deazaadenosine, S-adenosylhomocysteine or sinefungin into the cis solution markedly attenuated SAM-induced activation of RyR/Ca(2+) release channels. By Western blot analysis, arginine N-methyltransferase (PRMT1) and FK506 binding protein (FKBP) were detected in the SR used for reconstitution of RyR. In the presence of anti-PRMT1 antibody (1:100), SAM-induced activation of RyR/Ca(2+) channel was completely abolished. In addition, this SAM-induced increase in RyR/Ca(2+) channel activity was blocked by 30 microM ryanodine and by FK506 (100 microM), a ligand for the RyR accessory protein. These results suggest that protein methylation activates RyR/Ca(2+) release channels and may participate in the control of intracellular Ca(2+) mobilization in CASM cells by transferring a methyl group to the arginine moiety of the RyR accessory protein, FKBP 12. PMID:15118362

Chen, Ya-Fei; Zhang, Andrew Y; Zou, Ai-Ping; Campbell, William B; Li, Pin-Lan

2004-01-01

210

Intrinsically disordered proteins aggregate at fungal cell-to-cell channels and regulate intercellular connectivity  

PubMed Central

Like animals and plants, multicellular fungi possess cell-to-cell channels (septal pores) that allow intercellular communication and transport. Here, using a combination of MS of Woronin body-associated proteins and a bioinformatics approach that identifies related proteins based on composition and character, we identify 17 septal pore-associated (SPA) proteins that localize to the septal pore in rings and pore-centered foci. SPA proteins are not homologous at the primary sequence level but share overall physical properties with intrinsically disordered proteins. Some SPA proteins form aggregates at the septal pore, and in vitro assembly assays suggest aggregation through a nonamyloidal mechanism involving mainly ?-helical and disordered structures. SPA loss-of-function phenotypes include excessive septation, septal pore degeneration, and uncontrolled Woronin body activation. Together, our data identify the septal pore as a complex subcellular compartment and focal point for the assembly of unstructured proteins controlling diverse aspects of intercellular connectivity.

Lai, Julian; Koh, Chuan Hock; Tjota, Monika; Pieuchot, Laurent; Raman, Vignesh; Chandrababu, Karthik Balakrishna; Yang, Daiwen; Wong, Limsoon; Jedd, Gregory

2012-01-01

211

GTP-binding proteins mediate transmitter inhibition of voltage-dependent calcium channels  

Microsoft Academic Search

The modulation of voltage-dependent calcium channels by hormones and neurotransmitters has important implications for the control of many Ca2+-dependent cellular functions including exocytosis and contractility1-7. We made use of electrophysiological techniques, including whole-cell patch-clamp recordings from dorsal root ganglion (DRG) neurones, to demonstrate a role for GTP-binding proteins (G-proteins) as signal transducers in the noradrenaline- and gamma-aminobutyric acid (GABA)-induced inhibition

George G. Holz; Stanley G. Rane; Kathleen Dunlap

1986-01-01

212

Structure of the E. coli protein-conducting channel bound to a translating ribosome  

Microsoft Academic Search

Secreted and membrane proteins are translocated across or into cell membranes through a protein-conducting channel (PCC). Here we present a cryo-electron microscopy reconstruction of the Escherichia coli PCC, SecYEG, complexed with the ribosome and a nascent chain containing a signal anchor. This reconstruction shows a messenger RNA, three transfer RNAs, the nascent chain, and detailed features of both a translocating

Kakoli Mitra; Christiane Schaffitzel; Tanvir Shaikh; Florence Tama; Simon Jenni; Charles L. Brooks; Nenad Ban; Joachim Frank

2005-01-01

213

G-protein-gated TRP-like Cationic Channel Activated by Muscarinic Receptors  

PubMed Central

There is little information about the mechanisms by which G-protein–coupled receptors gate ion channels although many ionotropic receptors are well studied. We have investigated gating of the muscarinic cationic channel, which mediates the excitatory effect of acetylcholine in smooth muscles, and proposed a scheme consisting of four pairs of closed and open states. Channel kinetics appeared to be the same in cell-attached or outside-out patches whether the channel was activated by carbachol application or by intracellular dialysis with GTP?S. Since in the latter case G-proteins are permanently active, it is concluded that the cationic channel is the major determinant of its own gating, similarly to the KACh channel (Ivanova-Nikolova, T.T., and G.E. Breitwieser. 1997. J. Gen. Physiol. 109:245–253). Analysis of adjacent-state dwell times revealed connections between the states that showed features conserved among many other ligand-gated ion channels (e.g., nAChR, BKCa channel). Open probability (PO) of the cationic channel was increased by membrane depolarization consistent with the prominent U-shaped I-V relationship of the muscarinic whole-cell current at negative potentials. Membrane potential affected transitions within each closed-open state pair but had little effect on transitions between pairs; thus, the latter are likely to be caused by interactions of the channel with its ligands, e.g., Ca2+ and G?o-GTP. Channel activity was highly heterogeneous, as was evident from the prominent cycling behavior when PO was measured over 5-s intervals. This was related to the variable frequency of openings (as in the KACh channel) and, especially, to the number of long openings between consecutive long shuttings. Analysis of the underlying Markov chain in terms of probabilities allowed us to evaluate the contribution of each open state to the integral current (from shortest to longest open state: 0.1, 3, 24, and 73%) as PO increased 525-fold in three stages.

Zholos, Alexander V.; Zholos, Andrey A.; Bolton, Thomas B.

2004-01-01

214

Engineering of an E. coli outer membrane protein FhuA with increased channel diameter  

PubMed Central

Background Channel proteins like FhuA can be an alternative to artificial chemically synthesized nanopores. To reach such goals, channel proteins must be flexible enough to be modified in their geometry, i.e. length and diameter. As continuation of a previous study in which we addressed the lengthening of the channel, here we report the increasing of the channel diameter by genetic engineering. Results The FhuA ?1-159 diameter increase has been obtained by doubling the amino acid sequence of the first two N-terminal ?-strands, resulting in variant FhuA ?1-159 Exp. The total number of ?-strands increased from 22 to 24 and the channel surface area is expected to increase by ~16%. The secondary structure analysis by circular dichroism (CD) spectroscopy shows a high ?-sheet content, suggesting the correct folding of FhuA ?1-159 Exp. To further prove the FhuA ?1-159 Exp channel functionality, kinetic measurement using the HRP-TMB assay (HRP = Horse Radish Peroxidase, TMB = 3,3',5,5'-tetramethylbenzidine) were conducted. The results indicated a 17% faster diffusion kinetic for FhuA ?1-159 Exp as compared to FhuA ?1-159, well correlated to the expected channel surface area increase of ~16%. Conclusion In this study using a simple "semi rational" approach the FhuA ?1-159 diameter was enlarged. By combining the actual results with the previous ones on the FhuA ?1-159 lengthening a new set of synthetic nanochannels with desired lengths and diameters can be produced, broadening the FhuA ?1-159 applications. As large scale protein production is possible our approach can give a contribution to nanochannel industrial applications.

2011-01-01

215

Intra-membrane molecular interactions of K%2B channel proteins : application to problems in biodefense and bioenergy.  

SciTech Connect

Ion channel proteins regulate complex patterns of cellular electrical activity and ionic signaling. Certain K+ channels play an important role in immunological biodefense mechanisms of adaptive and innate immunity. Most ion channel proteins are oligomeric complexes with the conductive pore located at the central subunit interface. The long-term activity of many K+ channel proteins is dependent on the concentration of extracellular K+; however, the mechanism is unclear. Thus, this project focused on mechanisms underlying structural stability of tetrameric K+ channels. Using KcsA of Streptomyces lividans as a model K+ channel of known structure, the molecular basis of tetramer stability was investigated by: 1. Bioinformatic analysis of the tetramer interface. 2. Effect of two local anesthetics (lidocaine, tetracaine) on tetramer stability. 3. Molecular simulation of drug docking to the ion conduction pore. The results provide new insights regarding the structural stability of K+ channels and its possible role in cell physiology.

Moczydlowski, Edward G.

2013-07-01

216

The intracellular chloride ion channel protein CLIC1 undergoes a redox-controlled structural transition.  

PubMed

Most proteins adopt a well defined three-dimensional structure; however, it is increasingly recognized that some proteins can exist with at least two stable conformations. Recently, a class of intracellular chloride ion channel proteins (CLICs) has been shown to exist in both soluble and integral membrane forms. The structure of the soluble form of CLIC1 is typical of a soluble glutathione S-transferase superfamily protein but contains a glutaredoxin-like active site. In this study we show that on oxidation CLIC1 undergoes a reversible transition from a monomeric to a non-covalent dimeric state due to the formation of an intramolecular disulfide bond (Cys-24-Cys-59). We have determined the crystal structure of this oxidized state and show that a major structural transition has occurred, exposing a large hydrophobic surface, which forms the dimer interface. The oxidized CLIC1 dimer maintains its ability to form chloride ion channels in artificial bilayers and vesicles, whereas a reducing environment prevents the formation of ion channels by CLIC1. Mutational studies show that both Cys-24 and Cys-59 are required for channel activity. PMID:14613939

Littler, Dene R; Harrop, Stephen J; Fairlie, W Douglas; Brown, Louise J; Pankhurst, Greg J; Pankhurst, Susan; DeMaere, Matthew Z; Campbell, Terence J; Bauskin, Asne R; Tonini, Raffaella; Mazzanti, Michele; Breit, Samuel N; Curmi, Paul M G

2004-03-01

217

Inhibition of G Protein-Activated Inwardly Rectifying K+ Channels by Different Classes of Antidepressants  

Microsoft Academic Search

Various antidepressants are commonly used for the treatment of depression and several other neuropsychiatric disorders. In addition to their primary effects on serotonergic or noradrenergic neurotransmitter systems, antidepressants have been shown to interact with several receptors and ion channels. However, the molecular mechanisms that underlie the effects of antidepressants have not yet been sufficiently clarified. G protein-activated inwardly rectifying K+

Toru Kobayashi; Kazuo Washiyama; Kazutaka Ikeda

2011-01-01

218

Urea transporter UT3 functions as an efficient water channel. Direct evidence for a common water/urea pathway.  

PubMed

A family of molecular urea transporters (UTs) has been identified whose members appear to have an exceptionally high transport turnover rate. To test the hypothesis that urea transport involves passage through an aqueous channel, osmotic water permeability was measured in Xenopus oocytes expressing UTs. The UT3 class of urea transporters functioned as efficient water channels. Quantitative measurement of single channel water permeability (pf) using epitope-tagged rat UTs gave pf (in cm3/s x 10(-14)) of 0.14 +/- 0.11 (UT2) and 1.4 +/- 0.2 (UT3), compared with 6.0 and 2.3 for water channels AQP1 and AQP3, respectively. Relative single channel urea permeabilities (purea) were 1.0 (UT2), 0.44 (UT3), and 0.0 (AQP1). UT3-mediated water and urea transport were weakly temperature-dependent (activation energy <4 kcal/mol), inhibited > 75% by the urea transport inhibitor 1,3-dimethylthiourea, but not inhibited by the water transport inhibitor HgCl2. To test for a common water/urea pore, the urea reflection coefficient (sigmaurea) was measured by independent induced osmosis and solvent drag methods. In UT3-expressing oocytes, the time course of oocyte volume in response to different urea gradients (induced osmosis) gave sigmaurea approximately 0.3 for the UT3 pathway, in agreement with sigmaurea determined by the increase in uptake of [14C]urea during osmotic gradient-induced oocyte swelling (solvent drag). In oocytes of comparable water and urea permeability coexpressing AQP1 (permeable to water, not urea) and UT2 (permeable to urea, not water), sigmaurea = 1. These results indicate that UT3 functions as a urea/water channel utilizing a common aqueous pathway. The water transporting function and low urea reflection coefficient of UT3 in vasa recta may be important for the formation of a concentrated urine by countercurrent exchange in the kidney. PMID:9545259

Yang, B; Verkman, A S

1998-04-17

219

A membrane-delimited pathway of G-protein regulation of the guard-cell inward K+ channel.  

PubMed

GTP-binding protein (G-protein) regulation of inward rectifying K+ channels in the plasma membrane of Vicia (Vicia faba L.) guard cells has previously been demonstrated at the whole-cell level. However, whether a cytosolic signal transduction chain is required for G-protein regulation of K+ channels in Vicia guard cells, or in any plant cell type, remains unknown. In the present study, we assayed effects of several G-protein regulators on inward K+ channels in isolated inside-out membrane patches from Vicia guard cell protoplasts. Guanosine 5'-[gamma-thio]triphosphate, a nonhydrolyzable GTP analog that locks G proteins into their activated state, decreased the open state probability (Po) of single inward K+ channels. This decrease in Po was accompanied by an increase in one of the closed time constants of the K+ channel. Guanosine 5'-[beta-thio]diphosphate, a GDP analog that locks G proteins into their inactivated state, slightly increased the Po of the inward K+ channel and shortened the closed time constants. Pertussis toxin and cholera toxin, which ADP-ribosylate G proteins at different sites, decreased the Po of the inward K+ channel. Our data indicate that G proteins can act via a membrane-delimited pathway to regulate inward K+ channels in the guard-cell plasma membrane. PMID:8022777

Wu, W H; Assmann, S M

1994-07-01

220

K+ channel interactions detected by a genetic system optimized for systematic studies of membrane protein interactions  

PubMed Central

Organization of proteins into complexes is crucial for many cellular functions. However, most proteomic approaches primarily detect protein interactions for soluble proteins but are less suitable for membrane-associated complexes. Here we describe a mating-based split ubiquitin system (mbSUS) for systematic identification of interactions between membrane proteins as well as between membrane and soluble proteins. mbSUS allows in vivo cloning of PCR products into a vector set, detection of interactions via mating, regulated expression of baits, and improved selection of interacting proteins. Cloning is simplified by introduction of ? attachment sites for GATEWAY. Homo- and heteromeric interactions between Arabidopsis K+ channels KAT1, AKT1, and AKT2 were identified. Tests with deletion mutants demonstrate that the C terminus of KAT1 and AKT1 is necessary for physical assembly of complexes. Screening of a sorted collection of 84 plant proteins with K+ channels as bait revealed differences in oligomerization between KAT1, AKT1, and AtKC1, and allowed detection of putative interacting partners of KAT1 and AtKC1. These results show that mbSUS is suited for systematic analysis of membrane protein interactions.

Obrdlik, Petr; El-Bakkoury, Mohamed; Hamacher, Tanja; Cappellaro, Corinna; Vilarino, Cristina; Fleischer, Carola; Ellerbrok, Heinz; Kamuzinzi, Richard; Ledent, Valerie; Blaudez, Damien; Sanders, Dale; Revuelta, Jose L.; Boles, Eckhard; Andre, Bruno; Frommer, Wolf B.

2004-01-01

221

Nonuniform flow in compound channel: A 1-D method for assessing water level and discharge distribution  

NASA Astrophysics Data System (ADS)

This paper investigates 1-D modeling of nonuniform flows in compound channels. The issue is how to accurately predict both flow depth and mean velocity in the floodplain. A new model, called "Independent Subsections Method" (ISM), is presented here. Unlike classical 1-D models that solve a dynamic equation on the total cross section, the ISM estimates the water surface profile within each subsection. This enables the water level and the subsection mean velocities to be simultaneously calculated, without priority to any variable. In opposition to the Divided Channel Method (DCM), corrected DCM or the Exchange Discharge Model, the ISM assumes independent evolution of the discharge in each subsection of the compound channel. Indeed, this method does not assume equal head loss gradients in all subsections, and it does not impose the downstream discharge distribution. The ISM consists in a set of three coupled 1-D momentum equations (written within main channel, left-hand, and right-hand floodplains) and a mass conservation equation on the total cross section. Mass and momentum exchanges at the interfaces between subsections are explicitly accounted for. This method is validated against experimental data for developing flows in straight compound channel, flows in skewed compound channel, flows in a symmetric converging or diverging compound channel, and flows in an asymmetrical compound channel with an abrupt floodplain contraction. For the 46 runs, the ISM predicts flow depth and mean velocity in the floodplain with a maximum relative error of 8% and 19%, respectively. The ISM also appears to be a useful theoretical tool to improve our understanding of physical processes governing compound channel flows.

Proust, SéBastien; Bousmar, Didier; Riviere, Nicolas; Paquier, André; Zech, Yves

2009-12-01

222

Computational analysis of the soluble form of the intracellular chloride ion channel protein CLIC1.  

PubMed

The chloride intracellular channel (CLIC) family of proteins has the remarkable property of maintaining both a soluble form and an integral membrane form acting as an ion channel. The soluble form is structurally related to the glutathione-S-transferase family, and CLIC can covalently bind glutathione via an active site cysteine. We report approximately 0.6??s of molecular dynamics simulations, encompassing the three possible ligand-bound states of CLIC1, using the structure of GSH-bound human CLIC1. Noncovalently bound GSH was rapidly released from the protein, whereas the covalently ligand-bound protein remained close to the starting structure over 0.25??s of simulation. In the unliganded state, conformational changes in the vicinity of the glutathione-binding site resulted in reduced reactivity of the active site thiol. Elastic network analysis indicated that the changes in the unliganded state are intrinsic to the protein architecture and likely represent functional transitions. Overall, our results are consistent with a model of CLIC function in which covalent binding of glutathione does not occur spontaneously but requires interaction with another protein to stabilise the GSH binding site and/or transfer of the ligand. The results do not indicate how CLIC1 undergoes a radical conformational change to form a transmembrane chloride channel but further elucidate the mechanism by which CLICs are redox controlled. PMID:24089665

Jones, Peter M; Curmi, Paul M G; Valenzuela, Stella M; George, Anthony M

2013-01-01

223

Computational Analysis of the Soluble Form of the Intracellular Chloride Ion Channel Protein CLIC1  

PubMed Central

The chloride intracellular channel (CLIC) family of proteins has the remarkable property of maintaining both a soluble form and an integral membrane form acting as an ion channel. The soluble form is structurally related to the glutathione-S-transferase family, and CLIC can covalently bind glutathione via an active site cysteine. We report approximately 0.6??s of molecular dynamics simulations, encompassing the three possible ligand-bound states of CLIC1, using the structure of GSH-bound human CLIC1. Noncovalently bound GSH was rapidly released from the protein, whereas the covalently ligand-bound protein remained close to the starting structure over 0.25??s of simulation. In the unliganded state, conformational changes in the vicinity of the glutathione-binding site resulted in reduced reactivity of the active site thiol. Elastic network analysis indicated that the changes in the unliganded state are intrinsic to the protein architecture and likely represent functional transitions. Overall, our results are consistent with a model of CLIC function in which covalent binding of glutathione does not occur spontaneously but requires interaction with another protein to stabilise the GSH binding site and/or transfer of the ligand. The results do not indicate how CLIC1 undergoes a radical conformational change to form a transmembrane chloride channel but further elucidate the mechanism by which CLICs are redox controlled.

Jones, Peter M.; Curmi, Paul M. G.; Valenzuela, Stella M.; George, Anthony M.

2013-01-01

224

Channel  

NASA Technical Reports Server (NTRS)

[figure removed for brevity, see original site] Context image for PIA03693 Channel

This channel is located south of Iani Chaos.

Image information: VIS instrument. Latitude -10.9N, Longitude 345.5E. 17 meter/pixel resolution.

Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

2006-01-01

225

Vertical arrival structure of shipping noise in deep water channels  

Microsoft Academic Search

In passive sonar systems, knowledge of low-frequency shipping noise is significant for target detection performance. However, an accurate model for the shipping noise structure is difficult to obtain, because of the varying distributions of ships and complicated underwater environment. This work characterizes low-frequency distant shipping noise observed in deep water environments as a function of receiver depth and vertical arrival

Zizheng Li; Lisa M. Zurk; Barry Ma

2010-01-01

226

Mapping functional domains of chloride intracellular channel (CLIC) proteins in vivo.  

PubMed

Chloride intracellular channel (CLIC) proteins are small proteins distantly related to the omega family of glutathione S-transferases (GSTs). CLIC proteins are expressed in a wide variety of tissues in multicellular organisms and are targeted to specific cellular membranes. Members of this family are capable in vitro of changing conformation from a globular, soluble state to a membrane-inserted state in which they provide chloride conductance. The structural basis for in vivo CLIC protein function, however, is not well understood. We have mapped the functional domains of CLIC family members using an in vivo assay for membrane localization and function of CLIC proteins in the nematode Caenorhabditis elegans. A<70 amino acid N-terminal domain is a key determinant of membrane localization and function of invertebrate CLIC proteins. This domain, which we term the ''PTM'' domain, named after an amphipathic putative transmembrane helix contained within it, directs distinct C. elegans CLIC homologs to distinct subcellular membranes. We find that within the PTM region, the cysteine residues required for GST-type activity are unnecessary for invertebrate CLIC function, but that specific residues within the proposed transmembrane helix are necessary for correct targeting and protein function. We find that among all tested invertebrate CLIC proteins, function appears to be completely conserved despite striking differences in the charged residues contained within the amphipathic helix. This indicates that these residues do not contribute to anion selectivity as previously suggested. We find that outside the PTM region, the remaining three-quarters of CLIC protein sequence is functionally equivalent not only among vertebrate and invertebrate CLIC proteins, but also among the more distantly related GST-omega and GST-sigma proteins. The PTM region thus provides both targeting information and CLIC functional specificity, possibly adapting GST-type proteins to function as ion channels. PMID:16737711

Berry, Katherine L; Hobert, Oliver

2006-06-23

227

Capillary-Channeled Polymer (C-CP) Films as Processing Platforms for Protein Analysis by Matrix-Assisted Laser/Desorption Ionization Mass Spectrometry (MALDI-MS)  

NASA Astrophysics Data System (ADS)

Polypropylene (PP) capillary-channeled polymer (C-CP) films have parallel, ?m-sized channels that induce solution wicking via capillary action. Efficient mass transport from the solution phase to the channel surface leads to adsorption of hydrophobic protein solutes. The basic premise by which C-CP films can be used as media to manipulate analyte solutions (e.g., proteins in buffer), for the purpose of desalting or chromatographic separation prior to MALDI-MS analysis is presented here. Cytochrome c and myoglobin prepared in a Tris-HCl buffer, and ribonuclease A, lysozyme, and transferrin prepared in phosphate buffered saline (PBS), are used as the test solutions to demonstrate the desalting concept. Protein analysis is performed after deposition on a C-CP film with and without a water washing step, followed by spray deposition of a typical sinapinic acid matrix. Extracted MALDI mass spectra exhibit much improved signal-to-noise characteristics after water washing. A mixture of cytochrome c and myoglobin (2 ?L of 2.5 ?M each in Tris-HCl buffer) was applied, washed with water and spatially separated via simple capillary action (wicking) using a reversed-phase solvent composition of 0.1% trifluoroacetic acid (TFA) in 50:50 acetonitrile (ACN):H2O. Subsequent application of sinapinic acid followed by imaging of the film using MALDI-MS reveals that as the protein solution is wicked down the film, separation occurs.

Pittman, Jennifer J.; Manard, Benjamin T.; Kowalski, Paul J.; Marcus, R. Kenneth

2012-01-01

228

Capillary-channeled polymer (C-CP) films as processing platforms for protein analysis by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS).  

PubMed

Polypropylene (PP) capillary-channeled polymer (C-CP) films have parallel, ?m-sized channels that induce solution wicking via capillary action. Efficient mass transport from the solution phase to the channel surface leads to adsorption of hydrophobic protein solutes. The basic premise by which C-CP films can be used as media to manipulate analyte solutions (e.g., proteins in buffer), for the purpose of desalting or chromatographic separation prior to MALDI-MS analysis is presented here. Cytochrome c and myoglobin prepared in a Tris-HCl buffer, and ribonuclease A, lysozyme, and transferrin prepared in phosphate buffered saline (PBS), are used as the test solutions to demonstrate the desalting concept. Protein analysis is performed after deposition on a C-CP film with and without a water washing step, followed by spray deposition of a typical sinapinic acid matrix. Extracted MALDI mass spectra exhibit much improved signal-to-noise characteristics after water washing. A mixture of cytochrome c and myoglobin (2 ?L of 2.5 ?M each in Tris-HCl buffer) was applied, washed with water and spatially separated via simple capillary action (wicking) using a reversed-phase solvent composition of 0.1% trifluoroacetic acid (TFA) in 50:50 acetonitrile (ACN):H(2)O. Subsequent application of sinapinic acid followed by imaging of the film using MALDI-MS reveals that as the protein solution is wicked down the film, separation occurs. PMID:22012690

Pittman, Jennifer J; Manard, Benjamin T; Kowalski, Paul J; Marcus, R Kenneth

2012-01-01

229

Static and dynamic disorder in protein folding: experiments with single maltoporin channels  

NASA Astrophysics Data System (ADS)

The reversible binding of sugar to a single maltoporin channel allows us to study time and ensemble variations in the channel functional properties and interpret them using the language of static and dynamic disorder in protein folding. The channel is a trimer that is characterized by two primary parameters: the rate of sugar binding and the ion conductance. Time-resolved binding of maltohexasose molecules shows that whereas dynamic disorder -- the fluctuations in binding rate or in ionic conductance of a single trimer channel with time -- is relatively small, static disorder -- the heterogeneity of reaction rates or conductances among different trimers -- is highly pronounced. This heterogeneity suggests variations in maltoporin folding. The disorder in conductance shows no measurable correlation with the disorder in binding strength; variations in protein folding that are responsible for variations in protein folding that are responsible for variations in ionic conductance do not seem to affect sugar binding. We find 'cooperativity' in static disroder: conductances of monomers in the same trimer are closely similar compared to the range of possible conductances seen over an ensemble of trimers.

Kullman, Lisen; Winterhalter, Mathias; Bezrukov, Sergey M.

2003-05-01

230

Cytosolic Na+ Controls an Epithelial Na+ Channel Via the Go Guanine Nucleotide-Binding Regulatory Protein  

NASA Astrophysics Data System (ADS)

In tight Na+-absorbing epithelial cells, the rate of Na+ entry through amiloride-sensitive apical membrane Na+ channels is matched to basolateral Na+ extrusion so that cell Na+ concentration and volume remain steady. Control of this process by regulation of apical Na+ channels has been attributed to changes in cytosolic Ca2+ concentration or pH, secondary to changes in cytosolic Na+ concentration, although cytosolic Cl- seems also to be involved. Using mouse mandibular gland duct cells, we now demonstrate that increasing cytosolic Na+ concentration inhibits apical Na+ channels independent of changes in cytosolic Ca2+, pH, or Cl-, and the effect is blocked by GDP-? -S, pertussis toxin, and antibodies against the ? -subunits of guanine nucleotide-binding regulatory proteins (Go). In contrast, the inhibitory effect of cytosolic anions is blocked by antibodies to inhibitory guanine nucleotide-binding regulatory proteins (Gi1/Gi2. It thus appears that apical Na+ channels are regulated by Go and Gi proteins, the activities of which are controlled, respectively, by cytosolic Na+ and Cl-.

Komwatana, P.; Dinudom, A.; Young, J. A.; Cook, D. I.

1996-07-01

231

Molecular Determinants of Gem Protein Inhibition of P/Q-type Ca2+ Channels*  

PubMed Central

The RGK family of monomeric GTP-binding proteins potently inhibits high voltage-activated Ca2+ channels. The molecular mechanisms of this inhibition are largely unclear. In Xenopus oocytes, Gem suppresses the activity of P/Q-type Ca2+ channels on the plasma membrane. This is presumed to occur through direct interactions of one or more Gem inhibitory sites and the pore-forming Cav2.1 subunit in a manner dependent on the Ca2+ channel subunit ? (Cav?). In this study we investigated the molecular determinants in Gem that are critical for this inhibition. Like other RGK proteins, Gem contains a conserved Ras-like core and extended N and C termini. A 12-amino acid fragment in the C terminus was found to be crucial for and sufficient to produce Cav?-dependent inhibition, suggesting that this region forms an inhibitory site. A three-amino acid motif in the core was also found to be critical, possibly forming another inhibitory site. Mutating either site individually did not hamper Gem inhibition, but mutating both sites together completely abolished Gem inhibition without affecting Gem protein expression level or disrupting Gem interaction with Cav2.1 or Cav?. Mutating Gem residues that are crucial for interactions with previously demonstrated RGK modulators such as calmodulin, 14-3-3, and phosphatidylinositol lipids did not significantly affect Gem inhibition. These results suggest that Gem contains two candidate inhibitory sites, each capable of producing full inhibition of P/Q-type Ca2+ channels.

Fan, Mingming; Zhang, Wei K.; Buraei, Zafir; Yang, Jian

2012-01-01

232

Mechanical and conformational aspects of protein layers on water  

Microsoft Academic Search

Keywords: protein film, protein conformation, air\\/water interface, network formation, foam formation, foam stability, interfacial rheology, fracture behaviour.<\\/strong>The aim of this thesis was to obtain systematic information on the importance of mechanical and conformational aspects for the formation of a visco-elastic protein network at the air\\/water interface. Such a protein network is formed upon adsorption at the interface and is assumed

A. H. Martin

2003-01-01

233

2D IR spectroscopy reveals the role of water in the binding of channel-blocking drugs to the influenza M2 channel  

NASA Astrophysics Data System (ADS)

Water is an integral part of the homotetrameric M2 proton channel of the influenza A virus, which not only assists proton conduction but could also play an important role in stabilizing channel-blocking drugs. Herein, we employ two dimensional infrared (2D IR) spectroscopy and site-specific IR probes, i.e., the amide I bands arising from isotopically labeled Ala30 and Gly34 residues, to probe how binding of either rimantadine or 7,7-spiran amine affects the water dynamics inside the M2 channel. Our results show, at neutral pH where the channel is non-conducting, that drug binding leads to a significant increase in the mobility of the channel water. A similar trend is also observed at pH 5.0 although the difference becomes smaller. Taken together, these results indicate that the channel water facilitates drug binding by increasing its entropy. Furthermore, the 2D IR spectral signatures obtained for both probes under different conditions collectively support a binding mechanism whereby amantadine-like drugs dock in the channel with their ammonium moiety pointing toward the histidine residues and interacting with a nearby water cluster, as predicted by molecular dynamics simulations. We believe these findings have important implications for designing new anti-influenza drugs.

Ghosh, Ayanjeet; Wang, Jun; Moroz, Yurii S.; Korendovych, Ivan V.; Zanni, Martin; DeGrado, William F.; Gai, Feng; Hochstrasser, Robin M.

2014-06-01

234

Coupled Radon and Water Temperature Measurements to Characterize the Effects of Altered Stream Channel Planform  

NASA Astrophysics Data System (ADS)

In summer 2011, a 2.6 km reach of Meacham Creek, Oregon, USA, was altered from a straight, steep wall-based channel to more a sinuous, low-gradient channel. Key objectives of this restoration project were to increase the rate and magnitude of hyporheic exchange. The overarching goal was to initiate increased buffering and lagging of water temperature in the subsurface to mitigate warm surface water temperature in Meacham Creek, an important spawning and rearing stream for depressed populations of Chinook salmon and summer steelhead. To evaluate progress toward project goals and objectives, stream temperature and groundwater temperature in 22 wells have been measured hourly at the restoration site since March 2011. In addition, the radioactive isotope 222Rn was measured in each well and in the surface water on two occasions. The relative residence time of down welling stream water measured in the wells can be determined by ranked amplitude depression and lagged phase of annual temperature signals in the wells relative to that of the open channel flow. Residence times predicted by annual temperature signal dynamics are corroborated by 222Rn concentrations in each well. The data collected to date provide a foundation for developing a groundwater thermal model to predict the effects of channel reconfiguration on ground-surface water exchange and associated temperature effects at the reach scale.

Amerson, B. E.; Poole, G. C.; O'Daniel, S. J.

2013-12-01

235

TAT Fusion Protein Transduction into Isolated Mitochondria Is Accelerated by Sodium Channel Inhibitors†  

PubMed Central

Stringent control of ion and protein transport across the mitochondrial membranes is required to maintain mitochondrial function and biogenesis. In particular, the inner mitochondrial membrane is generally impermeable to proteins entering the matrix except via tightly regulated protein import mechanisms. Recently, cell penetrant peptides have been shown to move across the inner mitochondrial membrane in a manner suggesting an independent mechanism. HIV-1 transactivator of transcription (TAT) is an arginine-rich cell penetrant peptide, 47YGRKKRRQRRR57, which can transduce full-length proteins not only across the cell membrane but also into intracellular organelles. In this study, we investigated the ability of a TAT-containing protein to move into the mitochondrial matrix. Using a novel FACS assay for isolated, purified mitochondria, we show that TAT can deliver a modified fluorescent protein, mMDH-GFP, to the matrix of mitochondria and it is subsequently processed by the matrix peptidases. In addition, transduction of TAT-mMDH-GFP into mitochondria is independent of canonical protein import pathways as well as mitochondrial membrane potential. In direct contrast to published reports regarding the cell membrane where the sodium channel inhibitor, amiloride, blocks endocytosis and inhibits TAT transduction, TAT transduction into mitochondria is markedly increased by this same sodium channel inhibitor. These results confirm that the cell penetrant peptide, TAT, can readily transduce a protein cargo into the mitochondrial matrix. These results also demonstrate a novel role for mitochondrial sodium channels in mediating TAT transduction into mitochondria that is independent of endocytotic mechanisms. The mechanism of TAT transduction into mitochondria therefore is distinctly different from transduction across the cell membrane.

Rayapureddi, Jayanagendra P.; Tomamichel, Wendy J.; Walton, Sonia T.; Payne, R. Mark

2010-01-01

236

TAT fusion protein transduction into isolated mitochondria is accelerated by sodium channel inhibitors.  

PubMed

Stringent control of ion and protein transport across the mitochondrial membranes is required to maintain mitochondrial function and biogenesis. In particular, the inner mitochondrial membrane is generally impermeable to proteins entering the matrix except via tightly regulated protein import mechanisms. Recently, cell penetrant peptides have been shown to move across the inner mitochondrial membrane in a manner suggesting an independent mechanism. HIV-1 transactivator of transcription (TAT) is an arginine-rich cell penetrant peptide, 47YGRKKRRQRRR57, which can transduce full-length proteins not only across the cell membrane but also into intracellular organelles. In this study, we investigated the ability of a TAT-containing protein to move into the mitochondrial matrix. Using a novel FACS assay for isolated, purified mitochondria, we show that TAT can deliver a modified fluorescent protein, mMDH-GFP, to the matrix of mitochondria and it is subsequently processed by the matrix peptidases. In addition, transduction of TAT-mMDH-GFP into mitochondria is independent of canonical protein import pathways as well as mitochondrial membrane potential. In direct contrast to published reports regarding the cell membrane where the sodium channel inhibitor, amiloride, blocks endocytosis and inhibits TAT transduction, TAT transduction into mitochondria is markedly increased by this same sodium channel inhibitor. These results confirm that the cell penetrant peptide, TAT, can readily transduce a protein cargo into the mitochondrial matrix. These results also demonstrate a novel role for mitochondrial sodium channels in mediating TAT transduction into mitochondria that is independent of endocytotic mechanisms. The mechanism of TAT transduction into mitochondria therefore is distinctly different from transduction across the cell membrane. PMID:20925426

Rayapureddi, Jayanagendra P; Tomamichel, Wendy J; Walton, Sonia T; Payne, R Mark

2010-11-01

237

Aplysia synapse associated protein (APSAP): identification, characterization, and selective interactions with Shaker-type potassium channels.  

PubMed

The vertebrate post-synaptic density (PSD) is a region of high molecular complexity in which dynamic protein interactions modulate receptor localization and synaptic function. Members of the membrane-associated guanylate kinase (MAGUK) family of proteins represent a major structural and functional component of the vertebrate PSD. In order to investigate the expression and significance of orthologous PSD components associated with the Aplysia sensory neuron-motor neuron synapse, we have cloned an Aplysia Dlg-MAGUK protein, which we identify as Aplysia synapse associated protein (ApSAP). As revealed by western blot, RT-PCR, and immunocytochemical analyses, ApSAP is predominantly expressed in the CNS and is located in both sensory neuron and motor neurons. The overall amino acid sequence of ApSAP is 55-61% identical to Drosophila Dlg and mammalian Dlg-MAGUK proteins, but is more highly conserved within L27, PDZ, SH3, and guanylate kinase domains. Because these conserved domains mediate salient interactions with receptors and other PSD components of the vertebrate synapse, we performed a series of GST pull-down assays using recombinant C-terminal tail proteins from various Aplysia receptors and channels containing C-terminal PDZ binding sequences. We have found that ApSAP selectively binds to an Aplysia Shaker-type channel AKv1.1, but not to (i) NMDA receptor subunit AcNR1-1, (ii) potassium channel AKv5.1, (iii) receptor tyrosine kinase ApTrkl, (iv) glutamate receptor ApGluR1/4, (v) glutamate receptor ApGluR2/3, or (vi) glutamate receptor ApGluR7. These findings provide preliminary information regarding the expression and interactions of Dlg-MAGUK proteins of the Aplysia CNS, and will inform questions aimed at a functional analysis of how interactions in a protein network such as the PSD may regulate synaptic strength. PMID:18182049

Reissner, Kathryn J; Boyle, Heather D; Ye, Xiaojing; Carew, Thomas J

2008-05-01

238

Numerical sensitivity analysis of passive EHF and SMMW channels to tropospheric water vapor, clouds, and precipitation  

NASA Technical Reports Server (NTRS)

Potential uses of specific extremely High Frequency (EHF) and Sub-Millimeter-Wave (SMMW) channels at 90, 166, 183, 220, 325, 340, and 410 GHz for passive spaceborne remote sensing of the troposphere and lower stratosphere are investigated using an iterative numerical radiative transfer model. Collectively, these channels offer potential for high spatial resolution imaging using diffraction-limited apertures of practical size, along with the ability to profile water vapor, map precipitation beneath optically opaque cloud cover, and to measure nonprecipitating cloud (e.g., cirrus) parameters. A widely-spaced set of EHF and SMMW channels can yield observable degrees of freedom related to clouds and precipitation not available by exclusively using the more thoroughly studied microwave channels below 183 GHz. A new passive airborne imaging instrument for tropospheric meteorological sensing is described.

Gasiewski, A. J.

1992-01-01

239

Activation of protein kinase C differentially modulates neuronal Na+, Ca2+, and gamma-aminobutyrate type A channels.  

PubMed Central

Xenopus oocytes were used to study the interaction of neuronal quisqualate receptors with neuronal ion channels. Total mRNA was isolated from chick forebrain and injected into Xenopus oocytes. This technique led to the expression of functional voltage-gated Na+ and Ca2+ channels, of ligand-gated gamma-aminobutyrate and kainate receptor channels, and of quisqualate receptors that could activate endogenous chloride channels by means of inositol trisphosphate-mediated Ca2+ release. Exposure of the oocytes to quisqualate decreased the amplitude of the Na+ current and of the gamma-aminobutyrate type A-gated current and increased the amplitude of the Ba2+ current through Ca2+ channels. This modulation of neuronal ion channels by quisqualate could be mimicked by the protein kinase C activator phorbol 12-myristate 13-acetate and the diacylglycerol analogue 1,2-oleoylacetylglycerol. The kainate-gated channel was not affected by these agents. Phorbol esters that do not activate protein kinase C, alpha-phorbol 12-myristate 13-acetate and alpha-phorbol, were without effect. The inhibitor of protein kinase C, tamoxifen, prevented the modulatory effects of phorbol 12-myristate 13-acetate. The present evidence suggests that the activity of the neuronal Na+ and Ca2+ channels and the ligand-gated gamma-aminobutyrate type A receptor channel are under the control of protein kinase C and that neurotransmitters that activate protein kinase C could profoundly affect neuronal signaling.

Sigel, E; Baur, R

1988-01-01

240

Basolateral targeting and microtubule-dependent transcytosis of the aquaporin-2 water channel  

PubMed Central

The aquaporin-2 (AQP2) water channel relocates mainly to the apical plasma membrane of collecting duct principal cells after vasopressin (VP) stimulation. AQP2 transport to this membrane domain is assumed to be a direct route involving recycling of intracellular vesicles. However, basolateral plasma membrane expression of AQP2 is observed in vivo in principal cells. Here, we asked whether there is a transcytotic pathway of AQP2 trafficking between apical and basolateral membranes. We used MDCK cells in which AQP2 normally accumulates apically after VP exposure. In contrast, both site-specific biotinylation and immunofluorescence showed that AQP2 is strongly accumulated in the basolateral membrane, along with the endocytic protein clathrin, after a brief cold shock (4°C). This suggests that AQP2 may be constitutively targeted to basolateral membranes and then retrieved by clathrin-mediated endocytosis at physiological temperatures. Rab11 does not accumulate in basolateral membranes after cold shock, suggesting that the AQP2 in this location is not associated with Rab11-positive vesicles. After rewarming (37°C), basolateral AQP2 staining is diminished and it subsequently accumulates at the apical membrane in the presence of VP/forskolin, suggesting that transcytosis can be followed by apical insertion of AQP2. This process is inhibited by treatment with colchicine. Our data suggest that the cold shock procedure reveals the presence of microtubule-dependent AQP2 transcytosis, which represents an indirect pathway of apical AQP2 delivery in these cells. Furthermore, our data indicate that protein polarity data obtained from biotinylation assays, which require cells to be cooled to 4°C during the labeling procedure, should be interpreted with caution.

Yui, Naofumi; Lu, Hua A. J.; Chen, Ying; Nomura, Naohiro; Bouley, Richard

2013-01-01

241

Computation of water-surface profiles in open channels  

USGS Publications Warehouse

The standard step-backwater method of computing water-surface profiles is described in this chapter. The hydraulic principles and assumptions are reviewed, and the field data requirements are described. Certain special cases of backwater curves and certain special field conditions are discussed in detail. The technique is used to establish or extend stage-discharge ratings; to define areas which will be innundated by flood flows of a given frequency; and to compute profiles through various reaches, including multichannel flows, and past control structures such as bridges, culverts, and road embankments. A brief description of analysis of floodways and effects of encroachments is also presented.

Davidian, Jacob

1984-01-01

242

Ultrafast viscous water flow through nanostrand-channelled graphene oxide membranes  

NASA Astrophysics Data System (ADS)

Pressure-driven ultrafiltration membranes are important in separation applications. Advanced filtration membranes with high permeance and enhanced rejection must be developed to meet rising worldwide demand. Here we report nanostrand-channelled graphene oxide ultrafiltration membranes with a network of nanochannels with a narrow size distribution (3-5?nm) and superior separation performance. This permeance offers a 10-fold enhancement without sacrificing the rejection rate compared with that of graphene oxide membranes, and is more than 100 times higher than that of commercial ultrafiltration membranes with similar rejection. The flow enhancement is attributed to the porous structure and significantly reduced channel length. An abnormal pressure-dependent separation behaviour is also reported, where the elastic deformation of nanochannels offers tunable permeation and rejection. The water flow through these hydrophilic graphene oxide nanochannels is identified as viscous. This nanostrand-channelling approach is also extendable to other laminate membranes, providing potential for accelerating separation and water-purification processes.

Huang, Hubiao; Song, Zhigong; Wei, Ning; Shi, Li; Mao, Yiyin; Ying, Yulong; Sun, Luwei; Xu, Zhiping; Peng, Xinsheng

2013-12-01

243

A large iris-like expansion of a mechanosensitive channel protein induced by membrane tension  

NASA Technical Reports Server (NTRS)

MscL, a bacterial mechanosensitive channel of large conductance, is the first structurally characterized mechanosensor protein. Molecular models of its gating mechanisms are tested here. Disulfide crosslinking shows that M1 transmembrane alpha-helices in MscL of resting Escherichia coli are arranged similarly to those in the crystal structure of MscL from Mycobacterium tuberculosis. An expanded conformation was trapped in osmotically shocked cells by the specific bridging between Cys 20 and Cys 36 of adjacent M1 helices. These bridges stabilized the open channel. Disulfide bonds engineered between the M1 and M2 helices of adjacent subunits (Cys 32-Cys 81) do not prevent channel gating. These findings support gating models in which interactions between M1 and M2 of adjacent subunits remain unaltered while their tilts simultaneously increase. The MscL barrel, therefore, undergoes a large concerted iris-like expansion and flattening when perturbed by membrane tension.

Betanzos, Monica; Chiang, Chien-Sung; Guy, H. Robert; Sukharev, Sergei

2002-01-01

244

Hypomyelination and increased activity of voltage-gated K+ channels in mice lacking protein tyrosine phosphatase ?  

PubMed Central

Protein tyrosine phosphatase epsilon (PTP?) is strongly expressed in the nervous system; however, little is known about its physiological role. We report that mice lacking PTP? exhibit hypomyelination of sciatic nerve axons at an early post-natal age. This occurs together with increased activity of delayed- rectifier, voltage-gated potassium (Kv) channels and with hyperphosphorylation of Kv1.5 and Kv2.1 Kv channel ?-subunits in sciatic nerve tissue and in primary Schwann cells. PTP? markedly reduces Kv1.5 or Kv2.1 current amplitudes in Xenopus oocytes. Kv2.1 associates with a substrate-trapping mutant of PTP?, and PTP? profoundly reduces Src- or Fyn-stimulated Kv2.1 currents and tyrosine phosphorylation in transfected HEK 293 cells. In all, PTP? antagonizes activation of Kv channels by tyrosine kinases in vivo, and affects Schwann cell function during a critical period of Schwann cell growth and myelination.

Peretz, A.; Gil-Henn, H.; Sobko, A.; Shinder, V.; Attali, B.; Elson, A.

2000-01-01

245

A single WW domain is the predominant mediator of the interaction between the human ubiquitin-protein ligase Nedd4 and the human epithelial sodium channel.  

PubMed Central

The activity of the epithelial Na(+) channel (ENaC) is required for the maintenance of salt and water balance in the body. Channel activity is regulated by the ubiquitin-protein ligase Nedd4 ['neuronal precursor cell-expressed developmentally down-regulated (gene 4)'] that interacts with the channel via its WW domains. Mutations in channel subunits that disrupt this interaction cause Liddle's syndrome, a severe inherited form of hypertension. In previous studies we showed that WW domains 2, 3 and 4 of human Nedd4 bound to the human ENaC (hENaC) subunits, whereas WW domain 1 did not. Here we extend this observation to determine the binding affinities of the human Nedd4 WW domains for hENaC C-terminal peptides. We show that WW domains 2, 3 and 4 bind with differing affinities to Na(+) channel subunit peptides. WW domain 3 has the highest affinity and we predict that WW domain 3 contributes most of the binding because a construct containing the three WW domains bound no better than WW domain 3 alone. Further, a single amino acid change (Arg(165)-->Thr) in WW domain 1 enables binding to the alpha subunit of the channel to occur, with an affinity comparable with that of WW domain 4. Differential binding propensities between the various WW domains and Na(+) channel subunit peptides are explained on the basis of quantitative structural modelling of the complexes and their isolated components.

Lott, J Shaun; Coddington-Lawson, Sarah J; Teesdale-Spittle, Paul H; McDonald, Fiona J

2002-01-01

246

Detection and localization of a putative cyclic-GMP-activated channel protein in the protozoan ciliate Stentor coeruleus  

Microsoft Academic Search

Summary.  Immunoblotting and immunocytochemical assays were employed to identify and localize a channel protein activated by cyclic\\u000a GMP (cGMP) in the protozoan ciliate Stentor coeruleus. Analysis of whole-cell homogenate with antibodies raised against the ?-subunit of the cGMP-activated channel protein from\\u000a bovine rod outer segments and against cGMP revealed four major protein bands with molecular masses of 40 kDa, 63 kDa,

M. Walerczyk; H. Fabczak; S. Fabczak

2006-01-01

247

Biogas production from water hyacinth and channel grass used for phytoremediation of industrial effluents  

Microsoft Academic Search

The paper reports on the biogas production from water hyacinth (Eichhornia crassipes) and channel grass (Vallisneria spiralis) employed separately for phytoremediation of lignin and metal-rich pulp and paper mill and highly acidic distillery effluents. These plants eventually grow well in diluted effluent up to 40% (i.e., 2.5-times dilution with deionized water) and often take up metals and toxic materials from

V Singhal; J. P. N Rai

2003-01-01

248

Phase space analysis and classification of sonar echoes in shallow-water channels  

Microsoft Academic Search

A primary objective of active sonar systems is to detect, locate, and classify objects, such as mines, ships, and biologics, based on their sonar backscatter. A shallow-water ocean channel is a challenging environment in which to classify sonar echoes because interactions of the sonar signal with the ocean surface and bottom induce frequency-dependent changes (especially dispersion and damping) in the

Greg Okopal

2009-01-01

249

Regulation of the ABA-sensitive Arabidopsis potassium channel gene GORK in response to water stress  

Microsoft Academic Search

The phytohormone abscisic acid (ABA) regulates many stress-related processes in plants. In this context ABA mediates the responsiveness of plants to environmental stresses such as drought, cold or salt. In response to water stress, ABA induces stomatal closure by activating Ca2+, K+ and anion channels in guard cells. To understand the signalling pathways that regulate these turgor control elements, we

D. Becker; S. Hoth; P. Ache; S. Wenkel; M. R. G. Roelfsema; O. Meyerhoff; W. Hartung; R. Hedrich

2003-01-01

250

The nature of ion and water barrier crossings in a simulated ion channel.  

PubMed Central

Using a combination of techniques, including molecular dynamics, time-correlation analysis, stochastic dynamics, and fitting of continuum diffusion theory to electrophysiological data, a characterization is made of thermally driven sodium, water, and D2O motion within the gramicidin A channel. Since the channel contents are constrained to move in a single-file fashion, the motion that corresponds to experimentally measurable rates of permeation of the membrane is the motion of the center of mass of the channel contents. We therefore emphasize channel contents center-of-mass motion in our analysis of molecular dynamics computations. The usual free energy calculation techniques would be of questionable validity when applied to such motion. As an alternative to those techniques, we postulate a periodic sinusoidal free energy profile (related to the periodic structure of the helical channel) and deduce the fluid dynamic diffusion coefficient and the height and spacing of the free energy barriers from the form of the mean-square-deviation function, using stochastic computations. The fluid dynamic friction in each case appears similar to that for aqueous solution. However, the diffusive motions are modulated by a spatially periodic free energy profile with a periodicity characteristic of an L-D pair of amino acids in the gramicidin helix, approximately 1.7 A in the model we use. The barrier height depends on which substance is moving in the channel, but in each case is several times thermal energy. For barriers of this width and height, the motion is intermediate between the low-friction (transition-state) and high-friction (Brownian) limits. Thus, neither of these formalisms that have been used commonly to describe membrane permeation gives an accurate picture of the underlying physical process (although the Brownian description seems closer to correct). The non-Markovian Langevin equation must be solved to describe properly the statistics of the process. The "channel state of matter" characteristic of the channel contents appears to have some properties typical of the solid and some typical of the liquid state. The magnitude of the local friction and nature of the ion solvation are similar to the liquid state, but the periodicities of structure, free energy, and dynamics are somewhat solid-like. The alignment of water dipoles in the channel bears some resemblance to the orientational ordering of a nematic liquid crystal, but unlike a nematic liquid crystal, the waters have a degree of translational order as well. Thus, the "channel state" is not adequately described by analogy to either the solid or liquid states or to liquid crystals but must be dealt with as its own characteristic type of condensed matter.

Chiu, S. W.; Novotny, J. A.; Jakobsson, E.

1993-01-01

251

Expression of aquaporin water channels in the rat pituitary gland.  

PubMed

We investigated the expression of aquaporin (AQP) subtypes (AQP1-AQP9) in the rat pituitary gland by the reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. Specific amplification by RT-PCR was observed in AQP1, AQP3, AQP4 and AQP5 but not in the others. AQP1 immunoreactivity was detected in the endothelial cells of blood vessels. No positive reaction to AQP3 was observed by immunohistochemistry, in spite of the detection of mRNA. AQP4 was localized in the supporting cells of the adenohypophysis and neurohypophysis. AQP5 was expressed in parts of the marginal cells in Rathke's residual pouch. These observations suggest that several subtypes of the AQP family are involved in regulation of water homeostasis in the rat pituitary gland. PMID:18057834

Kuwahara, Sachi; Maeda, Seishi; Tanaka, Koichi; Hayakawa, Tetsu; Seki, Makoto

2007-11-01

252

Design of Peptide-Membrane Interactions to Modulate Single-File Water Transport through Modified Gramicidin Channels  

PubMed Central

Water permeability through single-file channels is affected by intrinsic factors such as their size and polarity and by external determinants like their lipid environment in the membrane. Previous computational studies revealed that the obstruction of the channel by lipid headgroups can be long-lived, in the range of nanoseconds, and that pore-length-matching membrane mimetics could speed up water permeability. To test the hypothesis of lipid-channel interactions modulating channel permeability, we designed different gramicidin A derivatives with attached acyl chains. By combining extensive molecular-dynamics simulations and single-channel water permeation measurements, we show that by tuning lipid-channel interactions, these modifications reduce the presence of lipid headgroups in the pore, which leads to a clear and selective increase in their water permeability.

Portella, Guillem; Polupanow, Tanja; Zocher, Florian; Boytsov, Danila A.; Pohl, Peter; Diederichsen, Ulf; de Groot, Bert L.

2012-01-01

253

Design of peptide-membrane interactions to modulate single-file water transport through modified gramicidin channels.  

PubMed

Water permeability through single-file channels is affected by intrinsic factors such as their size and polarity and by external determinants like their lipid environment in the membrane. Previous computational studies revealed that the obstruction of the channel by lipid headgroups can be long-lived, in the range of nanoseconds, and that pore-length-matching membrane mimetics could speed up water permeability. To test the hypothesis of lipid-channel interactions modulating channel permeability, we designed different gramicidin A derivatives with attached acyl chains. By combining extensive molecular-dynamics simulations and single-channel water permeation measurements, we show that by tuning lipid-channel interactions, these modifications reduce the presence of lipid headgroups in the pore, which leads to a clear and selective increase in their water permeability. PMID:23083713

Portella, Guillem; Polupanow, Tanja; Zocher, Florian; Boytsov, Danila A; Pohl, Peter; Diederichsen, Ulf; de Groot, Bert L

2012-10-17

254

Interannual variability of water mass properties in the Tunisia-Sicily Channel  

NASA Astrophysics Data System (ADS)

We analysed in situ CTD data collected on 36 joint Tunisian-Italian oceanographic cruises in the Tunisia-Sicily Channel (Cap Bon-Mazara del Vallo section) from 1995 to 2009 in order to identify the water masses in the region and to estimate the interannual variability of their hydrological characteristics. Besides the well-known AW (Atlantic Water) and LIW (Levantine Intermediate Water), other water masses have been identified though their presence is neither as steady nor as stable as the aforementioned ones. The WIW (Western Intermediate Water) flows beneath the AW towards the eastern basin while the IW (Ionian Water), with a highly intermittent character, flows within the subsurface layer towards the western basin. The flow of subsurface water (WIW and IW) is affected by intense mixing which modifies the water masses, tending to make them disappear. Moreover, the same hydrological time series produced interesting results concerning the increase in both temperature and salinity in the Tunisia-Sicily Channel. This trend especially concerns the deeper layers (LIW and tEMDW, i.e. transitional Eastern Mediterranean Deep Water) and we hypothesise that this is a direct response to climatic change occurring in the eastern basin.

Ben Ismail, Sana; Schroeder, Katrin; Sammari, Chérif; Gasparini, Gian Pietro; Borghini, Mireno; Aleya, Lotfi

2014-07-01

255

Bioinformatic Characterization of the Trimeric Intracellular Cation-Specific Channel Protein Family  

PubMed Central

Trimeric intracellular cation-specific (TRIC) channels are integral to muscle excitation–contraction coupling. TRIC channels provide counter-ionic flux when calcium is rapidly transported from intracellular stores to the cell cytoplasm. Until recently, knowledge of the presence of these proteins was limited to animals. We analyzed the TRIC family and identified a profusion of prokaryotic family members with topologies and motifs similar to those of their eukaryotic counterparts. Prokaryotic members far outnumber eukaryotic members, and although none has been functionally characterized, the evidence suggests that they function as secondary carriers. The presence of fused N- or C-terminal domains of known biochemical functions as well as genomic context analyses provide clues about the functions of these prokaryotic homologs. They are proposed to function in metabolite (e.g., amino acid/ nucleotide) efflux. Phylogenetic analysis revealed that TRIC channel homologs diverged relatively early during evolutionary history and that horizontal gene transfer was frequent in prokaryotes but not in eukaryotes. Topological analyses of TRIC channels revealed that these proteins possess seven putative transmembrane segments (TMSs), which arose by intragenic duplication of a three-TMS polypeptide-encoding genetic element followed by addition of a seventh TMS at the C terminus to give the precursor of all current TRIC family homologs. We propose that this family arose in prokaryotes.

Silverio, Abe L. F.

2014-01-01

256

Inhibition of Ca2+ Channels and Adrenal Catecholamine Release by G Protein Coupled Receptors  

PubMed Central

Catecholamines and other transmitters released from adrenal chromaffin cells play central roles in the “fight-or-flight” response and exert profound effects on cardiovascular, endocrine, immune, and nervous system function. As such, precise regulation of chromaffin cell exocytosis is key to maintaining normal physiological function and appropriate responsiveness to acute stress. Chromaffin cells express a number of different G protein coupled receptors (GPCRs) that sense the local environment and orchestrate this precise control of transmitter release. The primary trigger for catecholamine release is Ca2+ entry through voltage-gated Ca2+ channels, so it makes sense that these channels are subject to complex regulation by GPCRs. In particular G protein ?? heterodimers (G??) bind to and inhibit Ca2+ channels. Here I review the mechanisms by which GPCRs inhibit Ca2+ channels in chromaffin cells and how this might be altered by cellular context. This is related to the potent autocrine inhibition of Ca2+ entry and transmitter release seen in chromaffin cells. Recent data that implicate an additional inhibitory target of G?? on the exocytotic machinery and how this might fine tune neuroendocrine secretion are also discussed.

2011-01-01

257

Using Ion Channel-Forming Peptides to Quantify Protein-Ligand Interactions  

PubMed Central

This paper proposes a method for sensing affinity interactions by triggering disruption of self-assembly of ion channel-forming peptides in planar lipid bilayers. It shows that the binding of a derivative of alamethicin carrying a covalently attached sulfonamide ligand to carbonic anhydrase II (CA II) resulted in the inhibition of ion channel conductance through the bilayer. We propose that the binding of the bulky CA II protein (MW ~30 kD) to the ion channel-forming peptides (MW ~2.5 kD) either reduced the tendency of these peptides to self-assemble into a pore, or extracted them from the bilayer altogether. In both outcomes, the interactions between the protein and the ligand lead to a disruption of self-assembled pores. Addition of a competitive inhibitor – 4-carboxybenzenesulfonamide – to the solution released CA II from the alamethicin-sulfonamide conjugate and restored the current flow across the bilayer by allowing reassembly of the ion channels in the bilayer. Time-averaged recordings of the current over discrete time intervals made it possible to quantify this monovalent ligand binding interaction. This method gave a dissociation constant of ~2 µM for the binding of CA II to alamethicin-sulfonamide in the bilayer recording chamber: this value is consistent with a value obtained independently with CA II and a related sulfonamide derivative by isothermal titration calorimetry.

Mayer, Michael; Semetey, Vincent; Gitlin, Irina; Yang, Jerry; Whitesides, George M.

2008-01-01

258

An efficient method for modeling kinetic behavior of channel proteins in cardiomyocytes.  

PubMed

Characterization of the kinetic and conformational properties of channel proteins is a crucial element in the integrative study of congenital cardiac diseases. The proteins of the ion channels of cardiomyocytes represent an important family of biological components determining the physiology of the heart. Some computational studies aiming to understand the mechanisms of the ion channels of cardiomyocytes have concentrated on Markovian stochastic approaches. Mathematically, these approaches employ Chapman-Kolmogorov equations coupled with partial differential equations. As the scale and complexity of such subcellular and cellular models increases, the balance between efficiency and accuracy of algorithms becomes critical. We have developed a novel two-stage splitting algorithm to address efficiency and accuracy issues arising in such modeling and simulation scenarios. Numerical experiments were performed based on the incorporation of our newly developed conformational kinetic model for the rapid delayed rectifier potassium channel into the dynamic models of human ventricular myocytes. Our results show that the new algorithm significantly outperforms commonly adopted adaptive Runge-Kutta methods. Furthermore, our parallel simulations with coupled algorithms for multicellular cardiac tissue demonstrate a high linearity in the speedup of large-scale cardiac simulations. PMID:21576757

Wang, Chong; Beyerlein, Peter; Pospisil, Heike; Krause, Antje; Nugent, Chris; Dubitzky, Werner

2012-01-01

259

Evidence from oocyte expression that the erythrocyte water channel is distinct from band 3 and the glucose transporter.  

PubMed Central

It has been proposed that the mercurial-sensitive water transporter in mammalian erythrocytes is the anion exchanger band 3 (AE1) and/or the glucose transporter, band 4.5 (GLUT1). Using a functional assay for water channel expression in Xenopus oocytes (Zhang, R., K. A. Logee, and A. S. Verkman. 1990. J. Biol. Chem. 265:15375-15378), we compared osmotic water permeability (Pf) of oocytes injected with water, reticulocyte mRNA, AE1 mRNA, and GLUT1 mRNA. Injection of oocytes with 5-50 ng of in vitro-transcribed AE1 mRNA had no effect on Pf, but increased trans-stimulated 36Cl uptake greater than fourfold in a dinitro-disulfonic stilbene (DNDS)-inhibitable manner. Injection with 1-50 ng of in vitro-transcribed GLUT1 mRNA increased 3H-methylglucose uptake greater than 15-fold in a cytochalasin B-sensitive manner and increased Pf from (3.7 +/- 0.4) x 10(-4) cm/s (SE, n = 16, 10 degrees C) in water-injected oocytes up to (13 +/- 1) x 10(-4) cm/s (n = 18). Both the increments in sugar and water transport were inhibited by cytochalasin B (25 microM) and phloretin (0.2 mM); neither was inhibited by 0.3 mM HgCl2. In oocytes injected with 50 ng of rabbit reticulocyte mRNA, the Pf of (18 +/- 2) x 10(-4) cm/s (n = 18) was reduced to (4.0 +/- 0.6) x 10(-4) cm/s (n = 10) by HgCl2, but was not inhibited by DNDS (0.4 mM), cytochalasin B or phloretin. Coinjection of reticulocyte mRNA with antisense oligodeoxyribonucleotides against AE1 or GLUT1 did not affect Pf, but inhibited completely the incremental uptake of 36Cl or 3H-methylglucose, respectively. Expression of size-fractionated mRNA from reticulocyte gave a 2-2.5-kb size for water channel mRNA, less than the 4-4.5-kb size for the Cl transporter. These results provide evidence that facilitated water transport in erythrocytes is mediated not by bands 3 or 4.5, but by distinct water transport protein(s). Images

Zhang, R; Alper, S L; Thorens, B; Verkman, A S

1991-01-01

260

Differential modulation of Cav2.1 channels by calmodulin and Ca2+-binding protein 1  

PubMed Central

Cav2.1 channels, which mediate P/Q-type Ca2+ currents, undergo Ca2+/calmodulin (CaM)-dependent inactivation and facilitation that can significantly alter synaptic efficacy. Here we report that the neuronal Ca2+-binding protein 1 (CaBP1) modulates Cav2.1 channels in a manner that is markedly different from modulation by CaM. CaBP1 enhances inactivation, causes a depolarizing shift in the voltage dependence of activation, and does not support Ca2+-dependent facilitation of Cav2.1 channels. These inhibitory effects of CaBP1 do not require Ca2+, but depend on the CaM-binding domain in the ?1 subunit of Cav2.1 channels (?12.1). CaBP1 binds to the CaM-binding domain, co-immunoprecipitates with ?12.1 from transfected cells and brain extracts, and colocalizes with ?12.1 in discrete microdomains of neurons in the hippocampus and cerebellum. Our results identify an interaction between Ca2+ channels and CaBP1 that may regulate Ca2+-dependent forms of synaptic plasticity by inhibiting Ca2+ influx into neurons.

Lee, Amy; Westenbroek, Ruth E.; Haeseleer, Francoise; Palczewski, Krzysztof; Scheuer, Todd; Catterall, William A.

2006-01-01

261

An outer membrane channel protein of Mycobacterium tuberculosis with exotoxin activity.  

PubMed

The ability to control the timing and mode of host cell death plays a pivotal role in microbial infections. Many bacteria use toxins to kill host cells and evade immune responses. Such toxins are unknown in Mycobacterium tuberculosis. Virulent M. tuberculosis strains induce necrotic cell death in macrophages by an obscure molecular mechanism. Here we show that the M. tuberculosis protein Rv3903c (channel protein with necrosis-inducing toxin, CpnT) consists of an N-terminal channel domain that is used for uptake of nutrients across the outer membrane and a secreted toxic C-terminal domain. Infection experiments revealed that CpnT is required for survival and cytotoxicity of M. tuberculosis in macrophages. Furthermore, we demonstrate that the C-terminal domain of CpnT causes necrotic cell death in eukaryotic cells. Thus, CpnT has a dual function in uptake of nutrients and induction of host cell death by M. tuberculosis. PMID:24753609

Danilchanka, Olga; Sun, Jim; Pavlenok, Mikhail; Maueröder, Christian; Speer, Alexander; Siroy, Axel; Marrero, Joeli; Trujillo, Carolina; Mayhew, David L; Doornbos, Kathryn S; Muñoz, Luis E; Herrmann, Martin; Ehrt, Sabine; Berens, Christian; Niederweis, Michael

2014-05-01

262

Severe Acute Respiratory Syndrome Coronavirus Envelope Protein Ion Channel Activity Promotes Virus Fitness and Pathogenesis  

PubMed Central

Deletion of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) envelope (E) gene attenuates the virus. E gene encodes a small multifunctional protein that possesses ion channel (IC) activity, an important function in virus-host interaction. To test the contribution of E protein IC activity in virus pathogenesis, two recombinant mouse-adapted SARS-CoVs, each containing one single amino acid mutation that suppressed ion conductivity, were engineered. After serial infections, mutant viruses, in general, incorporated compensatory mutations within E gene that rendered active ion channels. Furthermore, IC activity conferred better fitness in competition assays, suggesting that ion conductivity represents an advantage for the virus. Interestingly, mice infected with viruses displaying E protein IC activity, either with the wild-type E protein sequence or with the revertants that restored ion transport, rapidly lost weight and died. In contrast, mice infected with mutants lacking IC activity, which did not incorporate mutations within E gene during the experiment, recovered from disease and most survived. Knocking down E protein IC activity did not significantly affect virus growth in infected mice but decreased edema accumulation, the major determinant of acute respiratory distress syndrome (ARDS) leading to death. Reduced edema correlated with lung epithelia integrity and proper localization of Na+/K+ ATPase, which participates in edema resolution. Levels of inflammasome-activated IL-1? were reduced in the lung airways of the animals infected with viruses lacking E protein IC activity, indicating that E protein IC function is required for inflammasome activation. Reduction of IL-1? was accompanied by diminished amounts of TNF and IL-6 in the absence of E protein ion conductivity. All these key cytokines promote the progression of lung damage and ARDS pathology. In conclusion, E protein IC activity represents a new determinant for SARS-CoV virulence.

Nieto-Torres, Jose L.; DeDiego, Marta L.; Verdia-Baguena, Carmina; Jimenez-Guardeno, Jose M.; Regla-Nava, Jose A.; Fernandez-Delgado, Raul; Castano-Rodriguez, Carlos; Alcaraz, Antonio; Torres, Jaume; Aguilella, Vicente M.; Enjuanes, Luis

2014-01-01

263

Channel Islands Deep Water Monitoring Plan Development Workshop Report, April 26-27, 2005. University of California, Santa Barbara.  

National Technical Information Service (NTIS)

In 2003, twelve marine protected areas were established in state waters (0-3 nmi) surrounding the Channel Islands. NOAA is considering extending this network (3-6 nmi) into deeper waters of the Channel Islands National Marine Sanctuary (CINMS). In order f...

D. Lott G. McFall K. Dalton S. Fangman S. Gittings

2005-01-01

264

Cotranslational Partitioning of Nascent Prion Protein into Multiple Populations at the Translocation Channel  

Microsoft Academic Search

The decisive events that direct a single polypeptide such as the prion protein (PrP) to be synthesized at the endoplasmic reticulum in both fully translocated and transmembrane forms are poorly understood. In this study, we demonstrate that the topological heterogeneity of PrP is determined cotranslationally, while at the translocation channel. By evaluating sequential inter- mediates during PrP topogenesis, we find

Soo Jung Kim; Ramanujan S. Hegde

2002-01-01

265

A large iris-like expansion of a mechanosensitive channel protein induced by membrane tension  

Microsoft Academic Search

MscL, a bacterial mechanosensitive channel of large conductance, is the first structurally characterized mechanosensor protein. Molecular models of its gating mechanisms are tested here. Disulfide crosslinking shows that M1 transmembrane ?-helices in MscL of resting Escherichia coli are arranged similarly to those in the crystal structure of MscL from Mycobacterium tuberculosis. An expanded conformation was trapped in osmotically shocked cells

Monica Betanzos; Chien-Sung Chiang; H. Robert Guy; Sergei Sukharev

2002-01-01

266

Alzheimer Disease Amyloid beta Protein Forms Calcium Channels in Bilayer Membranes: Blockade by Tromethamine and Aluminum  

Microsoft Academic Search

Amyloid beta protein (AbetaP) is the 40- to 42-residue polypeptide implicated in the pathogenesis of Alzheimer disease. We have incorporated this peptide into phosphatidylserine liposomes and then fused the liposomes with a planar bilayer. When incorporated into bilayers the AbetaP forms channels, which generate linear current-voltage relationships in symmetrical solutions. A permeability ratio, P_K\\/PCI, of 11 for the open AbetaP

Nelson Arispe; Eduardo Rojas; Harvey B. Pollard

1993-01-01

267

Conserved BK Channel-Protein Interactions Reveal Signals Relevant to Cell Death and Survival  

Microsoft Academic Search

The large-conductance Ca2+-activated K+ (BK) channel and its ?-subunit underlie tuning in non-mammalian sensory or hair cells, whereas in mammals its function is less clear. To gain insights into species differences and to reveal putative BK functions, we undertook a systems analysis of BK and BK-Associated Proteins (BKAPS) in the chicken cochlea and compared these results to other species. We

Bernd Sokolowski; Sandra Orchard; Margaret Harvey; Settu Sridhar; Yoshihisa Sakai

2011-01-01

268

TMEM16A Protein: A New Identity for Ca2+-Dependent Cl? Channels  

NSDL National Science Digital Library

Ca+-dependent Cl? channels (CaCCs) play a variety of physiological roles in different organs and tissues, including transepithelial Cl? secretion, smooth muscle contraction, regulation of neuronal excitability, and transduction of sensory stimuli. The recent identification of TMEM16A protein as an important component of CaCCs should allow a better understanding of their physiological role, structure-function relationship, and regulatory mechanisms.

Loretta Ferrera (Istituto Giannina Gaslini); Antonella Caputo (Istituto Giannina Gaslini); Luis Galietta (Istituto Giannina Gaslini)

2010-12-01

269

Isolation and characterization of channel catfish natural resistance associated macrophage protein gene  

Microsoft Academic Search

Natural resistance associated macrophage protein 1 (Nramp1) affects the ability of macrophages to kill pathogens. We cloned Nramp cDNA of channel catfish to identify potential molecular markers for disease resistance. Three different Nramp transcripts were identified: NrampCa-2912 nucleotides (nt), NrampCb-3245 nt, and NrampCc-3721 nt. At the 5? end, the transcripts have a common 2263 nt sequence containing the open reading

Hai Chen; Geoffrey C. Waldbieser; Charles D. Rice; Banu Elibol; William R. Wolters; Larry A. Hanson

2002-01-01

270

Differential Occurrence of Reluctant Openings in G-Protein-Inhibited N- and P/Q-Type Calcium Channels  

PubMed Central

Voltage-dependent inhibition of N- and P/Q-type calcium channels by G proteins is crucial for presynaptic inhibition of neurotransmitter release, and may contribute importantly to short-term synaptic plasticity. Such calcium-channel modulation could thereby impact significantly the neuro-computational repertoire of neural networks. The differential modulation of N and P/Q channels could even further enrich their impact upon synaptic tuning. Here, we performed in-depth comparison of the G-protein inhibition of recombinant N and P/Q channels, expressed in HEK 293 cells with the m2 muscarinic receptor. While both channel types display classic features of G-protein modulation (kinetic slowing of activation, prepulse facilitation, and voltage dependence of inhibition), we confirmed previously reported quantitative differences, with N channels displaying stronger inhibition and greater relief of inhibition by prepulses. A more fundamental, qualitative difference in the modulation of these two channels was revealed by a modified tail-activation paradigm, as well as by a novel “slope” analysis method comparing time courses of slow activation and prepulse facilitation. The stark contrast in modulatory behavior can be understood within the context of the “willing–reluctant” model, in which binding of G-protein ?? subunits to channels induces a reluctant mode of gating, where stronger depolarization is required for opening. Our experiments suggest that only N channels could be opened in the reluctant mode, at voltages normally spanned by neuronal action potentials. By contrast, P/Q channels appear to remain closed, especially over these physiological voltages. Further, the differential occurrence of reluctant openings is not explained by differences in the rate of G-protein unbinding from the two channels. These two scenarios predict very different effects of G-protein inhibition on the waveform of Ca2+ entry during action potentials, with potentially important consequences for the timing and efficacy of synaptic transmission.

Colecraft, Henry M.; Patil, Parag G.; Yue, David T.

2000-01-01

271

Protein-protein interactions and substrate channeling in orthologous and chimeric aldolase-dehydrogenase complexes.  

PubMed

Bacterial aldolase-dehydrogenase complexes catalyze the last steps in the meta cleavage pathway of aromatic hydrocarbon degradation. The aldolase (TTHB246) and dehydrogenase (TTHB247) from Thermus thermophilus were separately expressed and purified from recombinant Escherichia coli. The aldolase forms a dimer, while the dehydrogenase is a monomer; these enzymes can form a stable tetrameric complex in vitro, consisting of two aldolase and two dehydrogenase subunits. Upon complex formation, the K(m) value of 4-hydroxy-2-oxopentanoate, the substrate of TTHB246, is decreased 4-fold while the K(m) of acetaldehyde, the substrate of TTHB247, is increased 3-fold. The k(cat) values of each enzyme were reduced by ~2-fold when they were in a complex. The half-life of TTHB247 at 50 °C increased by ~4-fold when it was in a complex with TTHB246. The acetaldehyde product from TTHB246 could be efficiently channelled directly to TTHB247, but the channeling efficiency for the larger propionaldehyde was ~40% lower. A single A324G substitution in TTHB246 increased the channeling efficiency of propionaldehyde to a value comparable to that of acetaldehyde. Stable and catalytically competent chimeric complexes could be formed between the T. thermophilus enzymes and the orthologous aldolase (BphI) and dehydrogenase (BphJ) from the biphenyl degradation pathway of Burkholderia xenovorans LB400. However, channeling efficiencies for acetaldehyde in these chimeric complexes were ~10%. Structural and sequence analysis suggests that interacting residues in the interface of the aldolase-dehydrogenase complex are highly conserved among homologues, but coevolution of partner enzymes is required to fine-tune this interaction to allow for efficient substrate channeling. PMID:22316175

Baker, Perrin; Hillis, Colleen; Carere, Jason; Seah, Stephen Y K

2012-03-01

272

Highlights from recently determined structures of membrane proteins: a focus on channels and transporters  

PubMed Central

After decades of absent or lackluster growth, recent years have at long last witnessed an exponential growth in the number of novel membrane protein structures determined. Every single achievement has had a tremendous impact on the scientific community, typically providing an unprecedented wealth of information that only an atomic resolution structure can contribute to our molecular understanding of how a protein functions. Presented here is a review of some of the most exciting novel structures of channels and transporters determined by X-ray crystallography in the last two years, and a discussion of their analogies, differences and mechanistic implications.

Sciara, Giuliano; Mancia, Filippo

2012-01-01

273

Intracellular chloride channel protein CLIC1 regulates macrophage function through modulation of phagosomal acidification  

PubMed Central

Summary Intracellular chloride channel protein 1 (CLIC1) is a 241 amino acid protein of the glutathione S transferase fold family with redox- and pH-dependent membrane association and chloride ion channel activity. Whilst CLIC proteins are evolutionarily conserved in Metazoa, indicating an important role, little is known about their biology. CLIC1 was first cloned on the basis of increased expression in activated macrophages. We therefore examined its subcellular localisation in murine peritoneal macrophages by immunofluorescence confocal microscopy. In resting cells, CLIC1 is observed in punctate cytoplasmic structures that do not colocalise with markers for endosomes or secretory vesicles. However, when these macrophages phagocytose serum-opsonised zymosan, CLIC1 translocates onto the phagosomal membrane. Macrophages from CLIC1?/? mice display a defect in phagosome acidification as determined by imaging live cells phagocytosing zymosan tagged with the pH-sensitive fluorophore Oregon Green. This altered phagosomal acidification was not accompanied by a detectable impairment in phagosomal-lysosomal fusion. However, consistent with a defect in acidification, CLIC1?/? macrophages also displayed impaired phagosomal proteolytic capacity and reduced reactive oxygen species production. Further, CLIC1?/? mice were protected from development of serum transfer induced K/BxN arthritis. These data all point to an important role for CLIC1 in regulating macrophage function through its ion channel activity and suggest it is a suitable target for the development of anti-inflammatory drugs.

Jiang, Lele; Salao, Kanin; Li, Hui; Rybicka, Joanna M.; Yates, Robin M.; Luo, Xu Wei; Shi, Xin Xin; Kuffner, Tamara; Tsai, Vicky Wang-Wei; Husaini, Yasmin; Wu, Liyun; Brown, David A.; Grewal, Thomas; Brown, Louise J.; Curmi, Paul M. G.; Breit, Samuel N.

2012-01-01

274

Intracellular chloride channel protein CLIC1 regulates macrophage function through modulation of phagosomal acidification.  

PubMed

Intracellular chloride channel protein 1 (CLIC1) is a 241 amino acid protein of the glutathione S transferase fold family with redox- and pH-dependent membrane association and chloride ion channel activity. Whilst CLIC proteins are evolutionarily conserved in Metazoa, indicating an important role, little is known about their biology. CLIC1 was first cloned on the basis of increased expression in activated macrophages. We therefore examined its subcellular localisation in murine peritoneal macrophages by immunofluorescence confocal microscopy. In resting cells, CLIC1 is observed in punctate cytoplasmic structures that do not colocalise with markers for endosomes or secretory vesicles. However, when these macrophages phagocytose serum-opsonised zymosan, CLIC1 translocates onto the phagosomal membrane. Macrophages from CLIC1(-/-) mice display a defect in phagosome acidification as determined by imaging live cells phagocytosing zymosan tagged with the pH-sensitive fluorophore Oregon Green. This altered phagosomal acidification was not accompanied by a detectable impairment in phagosomal-lysosomal fusion. However, consistent with a defect in acidification, CLIC1(-/-) macrophages also displayed impaired phagosomal proteolytic capacity and reduced reactive oxygen species production. Further, CLIC1(-/-) mice were protected from development of serum transfer induced K/BxN arthritis. These data all point to an important role for CLIC1 in regulating macrophage function through its ion channel activity and suggest it is a suitable target for the development of anti-inflammatory drugs. PMID:22956539

Jiang, Lele; Salao, Kanin; Li, Hui; Rybicka, Joanna M; Yates, Robin M; Luo, Xu Wei; Shi, Xin Xin; Kuffner, Tamara; Tsai, Vicky Wang-Wei; Husaini, Yasmin; Wu, Liyun; Brown, David A; Grewal, Thomas; Brown, Louise J; Curmi, Paul M G; Breit, Samuel N

2012-11-15

275

Microfluidic Protein Patterning on Silicon Nitride Using Solvent Extracted Poly(dimethylsiloxane) Channels  

PubMed Central

Biomolecular patterning is essential for the creation of sensing motifs that rely on receptor-ligand binding for selectivity. Microfluidic devices have the potential to aid in the development of simple, robust methods for biomolecular patterning and therefore contribute to the generation of protein, DNA, and cell microarrays. In microfluidic patterning, the choice of both substrate and microfluidic channel material is essential for control of both the receptor binding for maximal signal generation as well as non-specific adsorption that acts as chemical noise. In this study, polystyrene, glass, silicon nitride, and poly(dimethylsiloxane) (PDMS) were evaluated as substrates for protein patterning using two types of PDMS microchannels for patterning, native PDMS and solvent-extracted PDMS (E-PDMS). E-PDMS microfluidic channels resulted in better patterning characteristics than native PDMS channels as determined by a higher fluorescence intensity of immobilized protein on all substrate types tested. Microfluidic patterning was then applied to perform two- and four-layer immunoassays.

He, Xinya; Dandy, David S.; Henry, Charles S.

2008-01-01

276

Manipulating L-type calcium channels in cardiomyocytes using split-intein protein transsplicing.  

PubMed

Manipulating expression of large genes (>6 kb) in adult cardiomyocytes is challenging because these cells are only efficiently transduced by viral vectors with a 4-7 kb packaging capacity. This limitation impedes understanding structure-function mechanisms of important proteins in heart. L-type calcium channels (LTCCs) regulate diverse facets of cardiac physiology including excitation-contraction coupling, excitability, and gene expression. Many important questions about how LTCCs mediate such multidimensional signaling are best resolved by manipulating expression of the 6.6 kb pore-forming ?1C-subunit in adult cardiomyocytes. Here, we use split-intein-mediated protein transsplicing to reconstitute LTCC ?1C-subunit from two distinct halves, overcoming the difficulty of expressing full-length ?1C in cardiomyocytes. Split-intein-tagged ?1C fragments encoding dihydropyridine-resistant channels were incorporated into adenovirus and reconstituted in cardiomyocytes. Similar to endogenous LTCCs, recombinant channels targeted to dyads, triggered Ca(2+) transients, associated with caveolin-3, and supported ?-adrenergic regulation of excitation-contraction coupling. This approach lowers a longstanding technical hurdle to manipulating large proteins in cardiomyocytes. PMID:24003157

Subramanyam, Prakash; Chang, Donald D; Fang, Kun; Xie, Wenjun; Marks, Andrew R; Colecraft, Henry M

2013-09-17

277

Manipulating L-type calcium channels in cardiomyocytes using split-intein protein transsplicing  

PubMed Central

Manipulating expression of large genes (>6 kb) in adult cardiomyocytes is challenging because these cells are only efficiently transduced by viral vectors with a 4–7 kb packaging capacity. This limitation impedes understanding structure–function mechanisms of important proteins in heart. L-type calcium channels (LTCCs) regulate diverse facets of cardiac physiology including excitation–contraction coupling, excitability, and gene expression. Many important questions about how LTCCs mediate such multidimensional signaling are best resolved by manipulating expression of the 6.6 kb pore-forming ?1C-subunit in adult cardiomyocytes. Here, we use split-intein–mediated protein transsplicing to reconstitute LTCC ?1C-subunit from two distinct halves, overcoming the difficulty of expressing full-length ?1C in cardiomyocytes. Split-intein–tagged ?1C fragments encoding dihydropyridine-resistant channels were incorporated into adenovirus and reconstituted in cardiomyocytes. Similar to endogenous LTCCs, recombinant channels targeted to dyads, triggered Ca2+ transients, associated with caveolin-3, and supported ?-adrenergic regulation of excitation–contraction coupling. This approach lowers a longstanding technical hurdle to manipulating large proteins in cardiomyocytes.

Subramanyam, Prakash; Chang, Donald D.; Fang, Kun; Xie, Wenjun; Marks, Andrew R.; Colecraft, Henry M.

2013-01-01

278

Shear strength and water resistance of modified soy protein adhesives  

Microsoft Academic Search

Soy protein polymers recently have been considered as alternatives to petroleum polymers to ease environmental pollution.\\u000a The use of soy proteins as adhesives for plywood has been limited because of their low water resistance. The objective of\\u000a this research was to test the water resistance of adhesives containing modified soy proteins in walnut, maple, poplar, and\\u000a pine plywood applications. Gluing

Xiuzhi Sun; Ke Bian

1999-01-01

279

Acylation of Membrane Proteins to Enhance Their Water Solubility  

Microsoft Academic Search

Membrane proteins are an integral part of the structure of membranes providing the functional aspect of the membrane with respect to the transport of water soluble molecules, signal recognition and reaction cataylsis. Primary, secondary and tertiary structure of these types of proteins warrant investigation in order to better understand their individual function. Both DNA sequencing and standard protein sequencing can

Robert Christopher Morton

1991-01-01

280

Acylation of membrane proteins to enhance their water solubility  

Microsoft Academic Search

Membrane proteins are an integral part of the structure of membranes providing the functional aspect of the membrane with respect to the transport of water soluble molecules, signal recognition and reaction catalysis. Primary, secondary and tertiary structure of these types of proteins warrant investigation in order to better understand their individual function. Both DNA sequencing and standard protein sequencing can

Robert Christopher Morton

1991-01-01

281

The water channel of cytochrome c oxidase: inferences from inhibitor studies.  

PubMed Central

Cytochrome c oxidase couples electron transfer to proton transfer from inside the mitochondrion to the cytosol. Protons pass through a channel; it is closed except when protons are pumped. Electron transfer is also coupled to a water cycle. Water moves into and out of the oxidase during electron transfer, presumably through a channel. The three processes are coupled because of the common dependence on electron transfer. If water and protons had to pass through the same channel for the proton to pass, it might be possible to block the pore by entraining small molecules in the flow. The data in this report indicate that there is a correlation between the ability of a compound to inhibit the oxidase and its size. Formamide and formaldehyde are potent inhibitors. Larger and smaller molecules are poor inhibitors. Formamide introduces an internal block in electron transfer. It is a slow-onset, reversible inhibitor, dependent on turnover to manifest its effects. Vesicular oxidase is less influenced by formamide than is soluble oxidase; formamide must pass a permeability barrier to act. The data are consistent with a proton channel with constrictions at both ends that open to yield a pore of approximately 4 A.

Kornblatt, J A

1998-01-01

282

Enantioselective Protein-Sterol Interactions Mediate Regulation of Both Prokaryotic and Eukaryotic Inward Rectifier K+ Channels by Cholesterol  

PubMed Central

Cholesterol is the major sterol component of all mammalian cell plasma membranes and plays a critical role in cell function and growth. Previous studies have shown that cholesterol inhibits inward rectifier K+ (Kir) channels, but have not distinguished whether this is due directly to protein-sterol interactions or indirectly to changes in the physical properties of the lipid bilayer. Using purified bacterial and eukaryotic Kir channels reconstituted into liposomes of controlled lipid composition, we demonstrate by 86Rb+ influx assays that bacterial Kir channels (KirBac1.1 and KirBac3.1) and human Kir2.1 are all inhibited by cholesterol, most likely by locking the channels into prolonged closed states, whereas the enantiomer, ent-cholesterol, does not inhibit these channels. These data indicate that cholesterol regulates Kir channels through direct protein-sterol interactions likely taking advantage of an evolutionarily conserved binding pocket.

D'Avanzo, Nazzareno; Hyrc, Krzysztof; Enkvetchakul, Decha; Covey, Douglas F.; Nichols, Colin G.

2011-01-01

283

G-protein mediates voltage regulation of agonist binding to muscarinic receptors: effects on receptor-Na/sup +/ channel interaction  

SciTech Connect

The authors previous experiments in membranes prepared from rat heart and brain led them to suggest that the binding of agonist to the muscarinic receptors and to the Na/sup +/ channels is a coupled event mediated by guanine nucleotide binding protein(s) (G-protein(s)). These in vitro findings prompted us to employ synaptoneurosomes from brain stem tissue to examine (i) the binding properties of (/sup 3/H) acetylcholine at resting potential and under depolarization conditions in the absence and presence of pertussis toxin; (ii) the binding of (/sup 3/H)batrachotoxin to Na/sup +/ channel(s) in the presence of the muscarinic agonists; and (iii) muscarinically induced /sup 22/Na/sup +/ uptake in the presence and absence of tetrodotoxin, which blocks Na/sup +/ channels. The findings indicate that agonist binding to muscarinic receptors is voltage dependent, that this process is mediated by G-protein(s), and that muscarinic agonists induce opening of Na/sup +/channels. The latter process persists even after pertussis toxin treatment, indicating that it is not likely to be mediated by pertussis toxin sensitive G-protein(s). The system with its three interacting components-receptor, G-protein, and Na/sup +/ channel-is such that at resting potential the muscarinic receptor induces opening of Na/sup +/ channels; this property may provide a possible physiological mechanism for the depolarization stimulus necessary for autoexcitation or repetitive firing in heart or brain tissues.

Cohen-Armon, M.; Garty, H.; Sokolovsky, M.

1988-01-12

284

Impact of Salt and Water on Protein Structural Dynamics  

NASA Astrophysics Data System (ADS)

Water is known as the lubricant of life. Without water, most proteins would lose their biological functions. Extensive studies have been carried out on how high concentration salts (dissolved in water) alter the stability and solubility of proteins. Such effects are thought to be mediated via salt-water interactions and water-protein interactions. This classic research field is known as the Hofmeister Series. We report the effects of Hofmeister Salts on the structural dynamics of proteins. Photoactive yellow protein (PYP), a bacterial blue light photoreceptor protein, is employed as a model system in this study. Time-resolved FTIR spectroscopic techniques allow us to probe the structural changes in proteins. Our data reveal that high concentration salt solutions alter the proton transfer pathway and suppress conformational changes in PYP upon photo-excitation. This study opens up a new dimension in the field of Hofmeister series. Further theoretical and experimental studies are needed in order to understand the dynamic properties of salt-water interactions and water-protein interactions.

Thubagere, Anu; Kelemen, Lorand; Nie, Beining; Kaledhonkar, Sandip; Xie, Aihua

2008-03-01

285

Constitutive and Regulated Membrane Expression of Aquaporin 1 and Aquaporin 2 Water Channels in Stably Transfected LLC-PK_1 Epithelial Cells  

Microsoft Academic Search

The aquaporins (AQPs) are a family of homologous water-channel proteins that can be inserted into epithelial cell plasma membranes either constitutively (AQP1) or by regulated exocytosis following vasopressin stimulation (AQP2). LLC-PK_1 porcine renal epithelial cells were stably transfected with cDNA encoding AQP2 (tagged with a C-terminal c-Myc epitope) or rat kidney AQP1 cDNA in an expression vector containing a cytomegalovirus

Toshiya Katsura; Jean-Marc Verbavatz; Javier Farinas; Tonghui Ma; Dennis A. Ausiello; A. S. Verkman; Dennis Brown

1995-01-01

286

Regulation of high-conductance anion channels by G proteins and 5-HT1A receptors in CHO cells.  

PubMed

This study addresses the mechanisms responsible for regulation of high-conductance anion channels by GTP binding proteins in Chinese hamster ovary (CHO) cells. Single-channel currents were measured in inside-out membrane patches using patch-clamp techniques. Anion-selective channels with a unitary conductance of 381 +/- 8 pS activated spontaneously in 48% of excised patches. In patches with no spontaneous channel activity, addition of GppNHp, a nonhydrolyzable analogue of GTP, activated channels in 8 of 12 studies, and in patches with spontaneous channel activity, GppNHp increased open probability in 4 of 4 experiments. In contrast, GDP beta S, a nonhydrolyzable GDP analogue, inhibited both spontaneous and GppNHp-induced channel activity. In patches without spontaneous channel activity, addition of cholera toxin activated channels in five of eight studies. Interestingly, pertussis toxin had a similar effect, activating channels in five of seven previously quiescent patches. To further evaluate the possible role of inhibitory G proteins in channel regulation, activity was measured in cell-attached patches in cells transfected with the serotonin 5-HT1A receptor, which is coupled to effector mechanisms through a pertussis toxin-sensitive G protein. Stimulation of 5-HT1A-transfected cells with the receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin caused a transient decrease in open probability in either standard or high-potassium solutions. In aggregate, these findings suggest that both cholera and pertussis toxin-sensitive G proteins contribute to regulation of high-conductance anion channels in CHO cells. PMID:7681262

Mangel, A W; Raymond, J R; Fitz, J G

1993-03-01

287

Classical Conditioning and Protein Kinase C Activation Regulate the Same Single Potassium Channel in Hermissenda crassicornis Photoreceptors  

Microsoft Academic Search

The patch-clamp technique was used to study the effects of classical conditioning and protein kinase C (PKC) activation on K^+ channels of identified neurons in the snail Hermissenda crassicornis. Here we present evidence that classical conditioning and PKC activation similarly modify the same K^+ channel. K^+ channels were recorded in cells from animals with different training experience. The 64-pS K^+

Rene Etcheberrigaray; Louis D. Matzel; I. Izja Lederhendler; Daniel L. Alkon

1992-01-01

288

Inhibition of a receptor-operated calcium channel in pig aortic microsomes by cyclic GMP-dependent protein kinase.  

PubMed Central

We have further characterized a putative receptor-operated Ca2+ channel that is activated by histamine and guanosine 5'-[beta gamma-imido]triphosphate. Insensitivity to verapamil, diltiazem or nicardipine, but inhibition by Ni2+ and SK&F 96365, further identify the channel with receptor-mediated Ca2+ entry in intact cells. Inhibition of the channel by cyclic-GMP-dependent protein kinase may contribute to vascular relaxation in response to nitrovasodilators.

Blayney, L M; Gapper, P W; Newby, A C

1991-01-01

289

Inhibition of a receptor-operated calcium channel in pig aortic microsomes by cyclic GMP-dependent protein kinase.  

PubMed

We have further characterized a putative receptor-operated Ca2+ channel that is activated by histamine and guanosine 5'-[beta gamma-imido]triphosphate. Insensitivity to verapamil, diltiazem or nicardipine, but inhibition by Ni2+ and SK&F 96365, further identify the channel with receptor-mediated Ca2+ entry in intact cells. Inhibition of the channel by cyclic-GMP-dependent protein kinase may contribute to vascular relaxation in response to nitrovasodilators. PMID:1705117

Blayney, L M; Gapper, P W; Newby, A C

1991-02-01

290

Water slug formation and motion in gas flow channels: the effects of geometry, surface wettability, and gravity.  

PubMed

Water emerging from ?100 ?m pores into millimeter-size gas flow channels forms drops that grow and become slugs which span the flow channel. Flowing gas causes the slugs to detach and move down the channel. The effect of channel geometry, surface wettability, and gravity on the formation and motion of water slugs has been analyzed using high-speed video images of the drops and differential pressure-time traces. Drops grow and appear, assuming a sequence of shapes that minimize the total interfacial energy of the gas-liquid and liquid-solid interfaces. The drops are initially spherical caps centered on the pore (the liquid contacts one wall). Above a certain size, the drops move to the corner, forming "corner drops" (the liquid contacts two walls). Corner drops grow across the channel, evolving into partial liquid bridges (drops confined by three walls), and finally the drops span the channel cross-section forming slugs (contacting all four walls). Smaller slugs are formed in channels with hydrophobic walls than in channels with hydrophilic walls. Smaller slugs are formed in channels with curved walls than in square or rectangular channels. Slugs move when the differential gas pressure overcomes the force to move the advancing and receding gas-liquid-solid contact lines of the slugs. Residual water left behind in corners by moving slugs reduces the barriers for drops to form slugs, causing the steady-state slug volumes to be smaller than those seen at start-up in dry channels. PMID:23876035

Cheah, May J; Kevrekidis, Ioannis G; Benziger, Jay B

2013-08-01

291

Inhibition of G Protein-Activated Inwardly Rectifying K+ Channels by Different Classes of Antidepressants  

PubMed Central

Various antidepressants are commonly used for the treatment of depression and several other neuropsychiatric disorders. In addition to their primary effects on serotonergic or noradrenergic neurotransmitter systems, antidepressants have been shown to interact with several receptors and ion channels. However, the molecular mechanisms that underlie the effects of antidepressants have not yet been sufficiently clarified. G protein-activated inwardly rectifying K+ (GIRK, Kir3) channels play an important role in regulating neuronal excitability and heart rate, and GIRK channel modulation has been suggested to have therapeutic potential for several neuropsychiatric disorders and cardiac arrhythmias. In the present study, we investigated the effects of various classes of antidepressants on GIRK channels using the Xenopus oocyte expression assay. In oocytes injected with mRNA for GIRK1/GIRK2 or GIRK1/GIRK4 subunits, extracellular application of sertraline, duloxetine, and amoxapine effectively reduced GIRK currents, whereas nefazodone, venlafaxine, mianserin, and mirtazapine weakly inhibited GIRK currents even at toxic levels. The inhibitory effects were concentration-dependent, with various degrees of potency and effectiveness. Furthermore, the effects of sertraline were voltage-independent and time-independent during each voltage pulse, whereas the effects of duloxetine were voltage-dependent with weaker inhibition with negative membrane potentials and time-dependent with a gradual decrease in each voltage pulse. However, Kir2.1 channels were insensitive to all of the drugs. Moreover, the GIRK currents induced by ethanol were inhibited by sertraline but not by intracellularly applied sertraline. The present results suggest that GIRK channel inhibition may reveal a novel characteristic of the commonly used antidepressants, particularly sertraline, and contributes to some of the therapeutic effects and adverse effects.

Kobayashi, Toru; Washiyama, Kazuo; Ikeda, Kazutaka

2011-01-01

292

The Proapoptotic Influenza A Virus Protein PB1-F2 Forms a Nonselective Ion Channel  

PubMed Central

Background PB1-F2 is a proapoptotic influenza A virus protein of approximately 90 amino acids in length that is located in the nucleus, cytosol and in the mitochondria membrane of infected cells. Previous studies indicated that the molecule destabilizes planar lipid bilayers and has a strong inherent tendency for multimerization. This may be correlate with its capacity to induce mitochondrial membrane depolarization. Methodology/Principal Findings Here, we investigated whether PB1-F2 is able to form ion channels within planar lipid bilayers and microsomes. For that purpose, a set of biologically active synthetic versions of PB1-F2 (sPB1-F2) derived from the IAV isolates A/Puerto Rico/8/34(H1N1) (IAVPR8), from A/Brevig Mission/1/1918(H1N1) (IAVSF2) or the H5N1 consensus sequence (IAVBF2) were used. Electrical and fluorimetric measurements show that all three peptides generate in planar lipid bilayers or in liposomes, respectively, a barely selective conductance that is associated with stochastic channel type fluctuations between a closed state and at least two defined open states. Unitary channel fluctuations were also generated when a truncated protein comprising only the 37 c-terminal amino acids of sPB1-F2 was reconstituted in bilayers. Experiments were complemented by extensive molecular dynamics simulations of the truncated fragment in a lipid bilayer. The results indicate that the c-terminal region exhibits a slightly bent helical fold, which is stable and remains embedded in the bilayer for over 180 ns. Conclusion/Significance The data support the idea that PB1-F2 is able to form protein channel pores with no appreciable selectivity in membranes and that the c-terminus is important for this function. This information could be important for drug development.

Henkel, Michael; Mitzner, David; Henklein, Peter; Meyer-Almes, Franz-Josef; Moroni, Anna; DiFrancesco, Mattia L.; Henkes, Leonhard M.; Kreim, Michael; Kast, Stefan M.; Schubert, Ulrich; Thiel, Gerhard

2010-01-01

293

Channel Incision and Water-Table Decline Along a Recently Formed Proglacial Stream, Mendenhall Valley, Southeastern Alaska  

USGS Publications Warehouse

Retreat of the Mendenhall Glacier, in southeastern Alaska, resulted in the formation of Mendenhall Lake, which has reduced the supply of coarse sediment to the proglacial Mendenhall River. Channel geometry surveys conducted in 1969 and 1998 over a 5.3 km reach of the Mendenhall River revealed reductions in mean bed elevations ranging from 0.4 to 1.5 meters based on cross sections replicated at 7 locations. Channel incision in the Mendenhall River is believed to be the result of a combination of factors resulting from localized and region-wide glacial retreat. In addition to a reduction of river stage due to channel incision, a decline in water-table elevations of about 0.6 m during a 17-year period from 1984 to 2001 was identified in an observation well located 250 m from the incising stream channel. Water-table elevations 600 m from the incising channel in the adjacent alluvial outwash aquifer respond in phase to changes in river stage, indicating water-levels in the adjacent aquifer are declining in response to river-channel incision. This study suggests channel incision can rapidly lower water-table elevations for large distances in the adjacent aquifer, potentially modifying the hydrology to a degree capable of influencing adjacent surface-water features, such as off-channel wetlands and flood-plain side channels.

Neal, Edward G.

2009-01-01

294

Stability analysis of a square rod bundle sub-channel in supercritical water reactor  

NASA Astrophysics Data System (ADS)

Extensive investigations on the flow and heat transfer behavior in SCWR fuel assembly have been undertaken worldwide. However, stability analysis of supercritical water in the sub-channels of tight lattices is still lacking. In this paper, the flow stability of a fuel bundle channel with square pitches has been analyzed using commercial CFD code-ANSYS Fluent. Typical dynamic instability of Density Wave Oscillation (DWO) has occurred in heated channel containing fluids at supercritical pressure. A further discussion about the impacts of various operational parameters (e.g. power input, system pressure, mass velocity, inlet temperature, etc) shows that the system becomes more stable as system pressure and/or mass flow rate increases. An increase in inlet temperature also has a stabilizing effect on the system.

Hai-jun, Wang; Ting, You; Lei, Zhang; Hong-fang, Gu; Yu-shan, Luo; Ji-lian, Bian

2013-07-01

295

River channel instability in East Anglia as a result of increasing water demand  

NASA Astrophysics Data System (ADS)

Both climate change and population growth are having an increasing effect on the morphodynamics of lowland rivers in East Anglia, mainly due to the rising water demand and the increasing magnitude of climate extremes such as droughts or floods. The region has had the UK's highest percentage increase in population in recent years and it is projected to rise by a further 20% over the next 15 years. East Anglia is also already the driest region in the UK. It receives only half of the national average annual rainfall in a normal year and most catchments are over-abstracted. The naturally-available water supply is low and therefore water has to be transferred from neighbouring catchments via pipelines and existing rivers, adding a significant amount of extra water to the natural river flows. Inadequate research is available to explain the spatial and temporal relationships of these additional flows on the affected river channels. A four year field study has been recently undertaken to explore the rates and causes of river channel instability on the River Stour in East Anglia. A river bank retreat of up to 1.3 m/year was recorded, which is much higher than the maximum rate of 0.2 m/year interpreted from an analysis of historical maps since 1886. The field study employed a unique combination of four geomorphologic field methods including the use of innovative photo-electronic erosion pins system for detailed continuous bank research. The studied river channel is used to transport additional water to supply, which was found to create 40% of all effective flows in the upstream reaches during the study period. The impact of this transferred water decreased downstream. The frequency of effective flows due to the water transfer scheme was examined against the river bank erosion retreat data considering the complexity of the channel boundary processes. Clear morphological evidence has also been collected that proves the effect that the water transfer flows are having on the river channel.

Anstead, Lenka; Tovey, Keith

2014-05-01

296

Heat transfer performance of Al2O3/water nanofluids in a mini channel heat sink.  

PubMed

The high density heat removal in electronic packaging is a challenging task of modern days. Finding compact, energy efficient and cost effective methods of heat removal is being the interest of researchers. In the present work, mini channel with forced convective heat transfer in simultaneously developing regime is investigated as the heat transfer coefficient is inversely proportional to hydraulic diameter. Mini channel heat sink is made from the aluminium plate of 30 mm square with 8 mm thickness. It has 15 mini channel of 0.9 mm width, 1.3 mm height and 0.9 mm of pitch. DI water and water based 0.1% and 0.2% volume fractions of Al2O3/water nanofluids are used as coolant. The flow rates of the coolants are maintained in such a way that it is simultaneously developing. Reynolds number is varied from 400 to 1600 and heat input is varied from 40 W to 70 W. The results showed that heat transfer coefficient is more than the heat transfer coefficient of fully developed flow. Also the heat transfer is more for nanofluids compared to DI water. PMID:24745233

Dominic, A; Sarangan, J; Suresh, S; Sai, Monica

2014-03-01

297

Bacillus subtilis spore protein SpoVAC functions as a mechanosensitive channel.  

PubMed

A critical event during spore germination is the release of Ca-DPA (calcium in complex with dipicolinic acid). The mechanism of release of Ca-DPA through the inner membrane of the spore is not clear, but proteins encoded by the Bacillus subtilis?spoVA operon are involved in the process. We cloned and expressed the spoVAC gene in Escherichia coli and characterized the SpoVAC protein. We show that SpoVAC protects E.?coli against osmotic downshift, suggesting that it might act as a mechanosensitive channel. Purified SpoVAC was reconstituted in unilamellar lipid vesicles to determine the gating mechanism and pore properties of the protein. By means of a fluorescence-dequenching assay, we show that SpoVAC is activated upon insertion into the membrane of the amphiphiles lysoPC and dodecylamine. Patch clamp experiments on E.?coli giant spheroplast as well as giant unilamellar vesicles (GUVs) containing SpoVAC show that the protein forms transient pores with main conductance values of about 0.15 and 0.1 nS respectively. Overall, our data indicate that SpoVAC acts as a mechanosensitive channel and has properties that would allow the release of Ca-DPA and amino acids during germination of the spore. PMID:24666282

Velásquez, Jeanette; Schuurman-Wolters, Gea; Birkner, Jan Peter; Abee, Tjakko; Poolman, Bert

2014-05-01

298

Long term regulation of cardiac L-type calcium channel by small G proteins.  

PubMed

Calcium ions are crucial elements of excitation-contraction coupling in cardiac myocytes. The intracellular Ca(2+ ) concentration changes continously during the cardiac cycle, but the Ca(2+ ) entering to the cell serves as an intracellular second messenger, as well. The Ca(2+ ) as a second messenger influences the activity of many intracellular signalling pathways and regulates gene expression. In cardiac myocytes the major pathway for Ca(2+ ) entry into cells is L-type calcium channel (LTCC). The precise control of LTCC function is essential for maintaining the calcium homeostasis of cardiac myocytes. Dysregulation of LTCC may result in different diseases like cardiac hypertrophy, arrhytmias, heart failure. The physiological and pathological structural changes in the heart are induced in part by small G proteins. These proteins are involved in wide spectrum of cell biological functions including protein transport, regulation of cell proliferation, migration, apoptosis, and cytoskeletal rearrangement. Understanding the crosstalk between small G proteins and LTCC may help to understand the pathomechanism of different cardiac diseases and to develop a new generation of genetically-encoded Ca(2+ ) channel inhibitors. PMID:21774757

Magyar, J; Jenes, A; Kistamás, K; Ruzsnavszky, F; Nánási, P P; Satin, J; Szentandrássy, N; Bányász, T

2011-01-01

299

Phase space analysis and classification of sonar echoes in shallow-water channels  

NASA Astrophysics Data System (ADS)

A primary objective of active sonar systems is to detect, locate, and classify objects, such as mines, ships, and biologics, based on their sonar backscatter. A shallow-water ocean channel is a challenging environment in which to classify sonar echoes because interactions of the sonar signal with the ocean surface and bottom induce frequency-dependent changes (especially dispersion and damping) in the signal as it propagates, the effects of which typically grow with range. Accordingly, the observed signal depends not only on the initial target backscatter, but also the propagation channel and how far the signal has propagated. These propagation effects can increase the variability of observed target echoes and degrade classification performance. Furthermore, uncertainty of the exact propagation channel and random variations within a channel cause classification features extracted from the received sonar echo to behave as random variables. With the goal of improving sonar signal classification in shallow-water environments, this work develops a phase space framework for studying sound propagation in channels with dispersion and damping. This approach leads to new moment features for classification that are invariant to dispersion and damping, the utility of which is demonstrated via simulation. In addition, the accuracy of a previously developed phase space approximation method for range-independent pulse propagation is analyzed and shown to be greater than the accuracy of the standard stationary phase approximation for both large and small times/distances. The phase space approximation is also extended to range dependent propagation. Finally, the phase space approximation is used to investigate the random nature of moment features for classification by calculating the moments of the moment features under uncertain and random channel assumptions. These moments of the moment features are used to estimate probability distribution functions for the moment features, and we explore several ways in which this information may be used to improve sonar classification performance.

Okopal, Greg

300

The p7 Protein of Hepatitis C Virus Forms Structurally Plastic, Minimalist Ion Channels  

PubMed Central

Hepatitis C virus (HCV) p7 is a membrane-associated oligomeric protein harboring ion channel activity. It is essential for effective assembly and release of infectious HCV particles and an attractive target for antiviral intervention. Yet, the self-assembly and molecular mechanism of p7 ion channelling are currently only partially understood. Using molecular dynamics simulations (aggregate time 1.2 µs), we show that p7 can form stable oligomers of four to seven subunits, with a bias towards six or seven subunits, and suggest that p7 self-assembles in a sequential manner, with tetrameric and pentameric complexes forming as intermediate states leading to the final hexameric or heptameric assembly. We describe a model of a hexameric p7 complex, which forms a transiently-open channel capable of conducting ions in simulation. We investigate the ability of the hexameric model to flexibly rearrange to adapt to the local lipid environment, and demonstrate how this model can be reconciled with low-resolution electron microscopy data. In the light of these results, a view of p7 oligomerization is proposed, wherein hexameric and heptameric complexes may coexist, forming minimalist, yet robust functional ion channels. In the absence of a high-resolution p7 structure, the models presented in this paper can prove valuable as a substitute structure in future studies of p7 function, or in the search for p7-inhibiting drugs.

Chandler, Danielle E.; Penin, Francois; Schulten, Klaus; Chipot, Christophe

2012-01-01

301

Voltage-dependent modulation of T-type calcium channels by protein tyrosine phosphorylation.  

PubMed Central

A T-type Ca2+ channel is expressed during differentiation of the male germ lineage in the mouse and is retained in sperm, where is it activated by contact with the the egg's extracellular matrix and controls sperm acrosomal exocytosis. Here, we examine the regulation of this Ca2+ channel in dissociated spermatogenic cells from the mouse using the whole-cell patch-clamp technique. T currents were enhanced, or facilitated, after strong depolarizations or high frequency stimulation. Voltage-dependent facilitation increased the Ca2+ current by an average of 50%. The same facilitation is produced by antagonists of protein tyrosine kinase activity. Conversely, antagonists of tyrosine phosphatase activity block voltage-dependent facilitation of the current. These data are consistent with the presence of a two-state model, in which T channels are maintained in a low (or zero) conductance state by tonic tyrosine phosphorylation and can be activated to a high conductance state by a tyrosine phosphatase activity. The positive and negative modulation of this channel by the tyrosine phosphorylation state provides a plausible mechanism for the control of sperm activity during the early stages of mammalian fertilization.

Arnoult, C; Lemos, J R; Florman, H M

1997-01-01

302

Transmembrane extension and oligomerization of the CLIC1 chloride intracellular channel protein upon membrane interaction.  

PubMed

Chloride intracellular channel proteins (CLICs) differ from most ion channels as they can exist in both soluble and integral membrane forms. The CLICs are expressed as soluble proteins but can reversibly autoinsert into the membrane to form active ion channels. For CLIC1, the interaction with the lipid bilayer is enhanced under oxidative conditions. At present, little evidence is available characterizing the structure of the putative oligomeric CLIC integral membrane form. Previously, fluorescence resonance energy transfer (FRET) was used to monitor and model the conformational transition within CLIC1 as it interacts with the membrane bilayer. These results revealed a large-scale unfolding between the C- and N-domains of CLIC1 as it interacts with the membrane. In the present study, FRET was used to probe lipid-induced structural changes arising in the vicinity of the putative transmembrane region of CLIC1 (residues 24-46) under oxidative conditions. Intramolecular FRET distances are consistent with the model in which the N-terminal domain inserts into the bilayer as an extended ?-helix. Further, intermolecular FRET was performed between fluorescently labeled CLIC1 monomers within membranes. The intermolecular FRET shows that CLIC1 forms oligomers upon oxidation in the presence of the membranes. Fitting the data to symmetric oligomer models of the CLIC1 transmembrane form indicates that the structure is large and most consistent with a model comprising approximately six to eight subunits. PMID:22082111

Goodchild, Sophia C; Angstmann, Christopher N; Breit, Samuel N; Curmi, Paul M G; Brown, Louise J

2011-12-20

303

Pentameric Assembly of Potassium Channel Tetramerization Domain-Containing Protein 5 (KCTD5)  

PubMed Central

We report the X-ray crystal structure of human potassium channel tetramerization domain-containing protein 5 (KCTD5), the first member of the family to be so characterized. Four findings were unexpected. First, the structure reveals assemblies of five subunits while tetramers were anticipated; pentameric stoichiometry is observed also in solution by scanning transmission electron microscopy mass analysis and analytical ultracentrifugation. Second, the same Bric-a-brac, Tramtrack, Broad Complex (BTB) domain surface mediates assembly of five KCTD5 and four voltage-gated potassium (Kv) channel subunits; four amino acid differences appear crucial. Third, KCTD5 complexes have well-defined N- and C-terminal modules separated by a flexible linker that swivels ~30°; the C-module shows a new fold and is required to bind Golgi re-assembling stacking protein 55 with ~1 ?M affinity as judged by surface plasmon resonance and ultracentrifugation. Fourth, despite the homology reflected in its name, KCTD5 does not impact operation of Kv4.2, Kv3.4, Kv2.1 or Kv1.2 channels.

Dementieva, Irina S.; Tereshko, Valentina; McCrossan, Zoe A.; Solomaha, Elena; Araki, Daniel; Xu, Chen; Grigorieff, Nikolaus; Goldstein, Steve A. N.

2009-01-01

304

Expression of G-protein inwardly rectifying potassium channels (GIRKs) in lung cancer cell lines  

PubMed Central

Background Previous data from our laboratory has indicated that there is a functional link between the ?-adrenergic receptor signaling pathway and the G-protein inwardly rectifying potassium channel (GIRK1) in human breast cancer cell lines. We wanted to determine if GIRK channels were expressed in lung cancers and if a similar link exists in lung cancer. Methods GIRK1-4 expression and levels were determined by reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR. GIRK protein levels were determined by western blots and cell proliferation was determined by a 5-bromo-2'-deoxyuridine (BrdU) assay. Results GIRK1 mRNA was expressed in three of six small cell lung cancer (SCLC) cell lines, and either GIRK2, 3 or 4 mRNA expression was detected in all six SCLC cell lines. Treatment of NCI-H69 with ?2-adrenergic antagonist ICI 118,551 (100 ?M) daily for seven days led to slight decreases of GIRK1 mRNA expression levels. Treatment of NCI-H69 with the ?-adrenergic agonist isoproterenol (10 ?M) decreased growth rates in these cells. The GIRK inhibitor U50488H (2 ?M) also inhibited proliferation, and this decrease was potentiated by isoproterenol. In the SCLC cell lines that demonstrated GIRK1 mRNA expression, we also saw GIRK1 protein expression. We feel these may be important regulatory pathways since no expression of mRNA of the GIRK channels (1 & 2) was found in hamster pulmonary neuroendocrine cells, a suggested cell of origin for SCLC, nor was GIRK1 or 2 expression found in human small airway epithelial cells. GIRK (1,2,3,4) mRNA expression was also seen in A549 adenocarcinoma and NCI-H727 carcinoid cell lines. GIRK1 mRNA expression was not found in tissue samples from adenocarcinoma or squamous cancer patients, nor was it found in NCI-H322 or NCI-H441 adenocarcinoma cell lines. GIRK (1,3,4) mRNA expression was seen in three squamous cell lines, GIRK2 was only expressed in one squamous cell line. However, GIRK1 protein expression was not seen in any non-SCLC cells. Conclusion We feel that this data may indicate that stimulation of GIRK1 or GIRK2 channels may be important in lung cancer. Stimulation of GIRK channels and ?-adrenergic signaling may activate similar signaling pathways in both SCLC and breast cancer, but lead to different results.

Plummer, Howard K; Dhar, Madhu S; Cekanova, Maria; Schuller, Hildegard M

2005-01-01

305

Reaction enthalpies along the two channels of geminate electron recombination in liquid-to-supercritical water  

NASA Astrophysics Data System (ADS)

Ionizing radiation or UV light produces electrons and H2O+ ions in water. These species transform into hydrated electron, e-aq, hydrated H3O+ ion, and ·OH radical in each other's neighborhood much faster than any forthcoming chemical transformation. Part of the electrons escapes their geminate partners. There exists two possible paths for the remaining fraction to react: H3O++e-aq=H3O· [channel (A)] and ·OH+e-aq=OH- [channel (B)]. We devised two thermodynamic cycles for the computation of the reaction enthalpies of both channels. Channel (A) was found to be endothermic with an enthalpy of 3.61 eV at room temperature. The enthalpy is seen to be almost constant up to 500 K, to increase at 600 K and to drop abruptly around 650 K, i.e. in the region where the dielectric constant is below 20. Channel (B) was found to be exothermic with an enthalpy of -2.33 eV at room temperature. It is becoming gradually less exothermic with increasing temperature the variation becoming fast around 650 K. The tendency of these thermochemical results parallel with recent kinetic calculations by Torres-Alacan et al. (J. Torres-Alacan, S. Kratz, P. Vöhringer, 2011. Phys. Chem. Chem. Phys. 13, 20806-20819)

Schiller, Robert; Horváth, Ákos

2013-11-01

306

Phosphorylation-Dependent and Phosphorylation-Independent Modes of Modulation of Shaker Family Voltage-Gated Potassium Channels by Src Family Protein Tyrosine Kinases  

Microsoft Academic Search

Modulation of voltage-gated potassium (Kv) channels by pro- tein phosphorylation plays an essential role in the regulation of the membrane properties of cells. Protein-protein binding do- mains, such as Src homology 3 (SH3) domains, direct ion channel modulation by coupling the channels with intracellular signaling enzymes. The conventional view is that protein kinase binding to ion channels leads to modulation

Michael N. Nitabach; D. Alberto Llamas; Ian J. Thompson; Kerry A. Collins; Todd C. Holmes

307

Water-deficit-responsive proteins in maritime pine.  

PubMed

Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to identify drought-responsive proteins during progressive water deprivation of two-year old maritime pine seedlings. Stress was applied by withholding water during vegetative growth. Needles were sampled before, during and after the stress. Out of about 1000 spots that were quantified by computer analysis, 38 responded during stress. Some proteins were accumulated while others were suppressed. One to three internal microsequences were obtained for 11 proteins, 10 of which were identified on the basis of sequence homologies. These proteins are quite diverse and are involved in photosynthesis, cell elongation, antioxidant metabolism and lignification. PMID:9747804

Costa, P; Bahrman, N; Frigerio, J M; Kremer, A; Plomion, C

1998-11-01

308

Structure-function of proteins interacting with the ?1 pore-forming subunit of high-voltage-activated calcium channels  

PubMed Central

Openings of high-voltage-activated (HVA) calcium channels lead to a transient increase in calcium concentration that in turn activate a plethora of cellular functions, including muscle contraction, secretion and gene transcription. To coordinate all these responses calcium channels form supramolecular assemblies containing effectors and regulatory proteins that couple calcium influx to the downstream signal cascades and to feedback elements. According to the original biochemical characterization of skeletal muscle Dihydropyridine receptors, HVA calcium channels are multi-subunit protein complexes consisting of a pore-forming subunit (?1) associated with four additional polypeptide chains ?, ?2, ?, and ?, often referred to as accessory subunits. Twenty-five years after the first purification of a high-voltage calcium channel, the concept of a flexible stoichiometry to expand the repertoire of mechanisms that regulate calcium channel influx has emerged. Several other proteins have been identified that associate directly with the ?1-subunit, including calmodulin and multiple members of the small and large GTPase family. Some of these proteins only interact with a subset of ?1-subunits and during specific stages of biogenesis. More strikingly, most of the ?1-subunit interacting proteins, such as the ?-subunit and small GTPases, regulate both gating and trafficking through a variety of mechanisms. Modulation of channel activity covers almost all biophysical properties of the channel. Likewise, regulation of the number of channels in the plasma membrane is performed by altering the release of the ?1-subunit from the endoplasmic reticulum, by reducing its degradation or enhancing its recycling back to the cell surface. In this review, we discuss the structural basis, interplay and functional role of selected proteins that interact with the central pore-forming subunit of HVA calcium channels.

Neely, Alan; Hidalgo, Patricia

2014-01-01

309

Relaxation channels of two-vibron bound states in alpha-helix proteins.  

PubMed

Relaxation channels for two-vibron bound states in an anharmonic alpha-helix protein are studied. According to a recently established small polaron model [V. Pouthier, Phys. Rev. E 68, 021909 (2003)], it is shown that the relaxation originates in the interaction between the dressed anharmonic vibrons and the remaining phonons. This interaction is responsible for the occurrence of transitions between two-vibron eigenstates mediated by both phonon absorption and phonon emission. At biological temperature, the relaxation rate does not significantly depend on the nature of the two-vibron states involved in the process. The lifetime for both bound and free states is of the same order of magnitude and ranges between 0.1 and 1.0 ps for realistic parameter values. By contrast, the relaxation channels strongly depend on the nature of the two-vibron states which is a consequence of the breatherlike behavior of the two-vibron bound states. PMID:15169042

Pouthier, V; Falvo, C

2004-04-01

310

Relaxation channels of two-vibron bound states in ?-helix proteins  

NASA Astrophysics Data System (ADS)

Relaxation channels for two-vibron bound states in an anharmonic ?-helix protein are studied. According to a recently established small polaron model [V. Pouthier, Phys. Rev. E 68, 021909 (2003)], it is shown that the relaxation originates in the interaction between the dressed anharmonic vibrons and the remaining phonons. This interaction is responsible for the occurrence of transitions between two-vibron eigenstates mediated by both phonon absorption and phonon emission. At biological temperature, the relaxation rate does not significantly depend on the nature of the two-vibron states involved in the process. The lifetime for both bound and free states is of the same order of magnitude and ranges between 0.1 and 1.0 ps for realistic parameter values. By contrast, the relaxation channels strongly depend on the nature of the two-vibron states which is a consequence of the breatherlike behavior of the two-vibron bound states.

Pouthier, V.; Falvo, C.

2004-04-01

311

Vasoconstrictor agonists activate G-protein-dependent receptor-operated calcium channels in pig aortic microsomes.  

PubMed Central

Receptor-operated Ca2+ channels were characterized by their ability to decrease steady-state ATP-dependent Ca2+ accumulation into pig aortic microsomes. The vasoconstrictor agents noradrenaline, angiotensin II and adenosine 5'-[alpha beta-methylene]triphosphate (pp[CH2]pA) all decreased Ca2+ accumulation only when sonicated into vesicles (to allow access to receptor sites) and in the presence of guanosine 5'-[beta gamma-imido]triphosphate to activate transducing G-proteins. The effect of noradrenaline was inhibited by the alpha 2 antagonist yohimbine, but not by the alpha 1 antagonist prazosin. The effect of none of the agonists was reversed by diltiazem. SK&F 96365 (an inhibitor of receptor-mediated Ca2+ influx into intact cells) reversed the effect of noradrenaline, but not that of pp[CH2]pA, which suggests that at least two receptor-operated channels may be present in this preparation.

Blayney, L M; Gapper, P W; Newby, A C

1992-01-01

312

Vasoconstrictor agonists activate G-protein-dependent receptor-operated calcium channels in pig aortic microsomes.  

PubMed

Receptor-operated Ca2+ channels were characterized by their ability to decrease steady-state ATP-dependent Ca2+ accumulation into pig aortic microsomes. The vasoconstrictor agents noradrenaline, angiotensin II and adenosine 5'-[alpha beta-methylene]triphosphate (pp[CH2]pA) all decreased Ca2+ accumulation only when sonicated into vesicles (to allow access to receptor sites) and in the presence of guanosine 5'-[beta gamma-imido]triphosphate to activate transducing G-proteins. The effect of noradrenaline was inhibited by the alpha 2 antagonist yohimbine, but not by the alpha 1 antagonist prazosin. The effect of none of the agonists was reversed by diltiazem. SK&F 96365 (an inhibitor of receptor-mediated Ca2+ influx into intact cells) reversed the effect of noradrenaline, but not that of pp[CH2]pA, which suggests that at least two receptor-operated channels may be present in this preparation. PMID:1347211

Blayney, L M; Gapper, P W; Newby, A C

1992-02-15

313

Ancient Origins of RGK Protein Function: Modulation of Voltage-Gated Calcium Channels Preceded the Protostome and Deuterostome Split.  

PubMed

RGK proteins, Gem, Rad, Rem1, and Rem2, are members of the Ras superfamily of small GTP-binding proteins that interact with Ca2+ channel ? subunits to modify voltage-gated Ca2+ channel function. In addition, RGK proteins affect several cellular processes such as cytoskeletal rearrangement, neuronal dendritic complexity, and synapse formation. To probe the phylogenetic origins of RGK protein-Ca2+ channel interactions, we identified potential RGK-like protein homologs in genomes for genetically diverse organisms from both the deuterostome and protostome animal superphyla. RGK-like protein homologs cloned from Danio rerio (zebrafish) and Drosophila melanogaster (fruit flies) expressed in mammalian sympathetic neurons decreased Ca2+ current density as reported for expression of mammalian RGK proteins. Sequence alignments from evolutionarily diverse organisms spanning the protostome/deuterostome divide revealed conservation of residues within the RGK G-domain involved in RGK protein - Cav? subunit interaction. In addition, the C-terminal eleven residues were highly conserved and constituted a signature sequence unique to RGK proteins but of unknown function. Taken together, these data suggest that RGK proteins, and the ability to modify Ca2+ channel function, arose from an ancestor predating the protostomes split from deuterostomes approximately 550 million years ago. PMID:24992013

Puhl, Henry L; Lu, Van B; Won, Yu-Jin; Sasson, Yehezkel; Hirsch, Joel A; Ono, Fumihito; Ikeda, Stephen R

2014-01-01

314

Ancient Origins of RGK Protein Function: Modulation of Voltage-Gated Calcium Channels Preceded the Protostome and Deuterostome Split  

PubMed Central

RGK proteins, Gem, Rad, Rem1, and Rem2, are members of the Ras superfamily of small GTP-binding proteins that interact with Ca2+ channel ? subunits to modify voltage-gated Ca2+ channel function. In addition, RGK proteins affect several cellular processes such as cytoskeletal rearrangement, neuronal dendritic complexity, and synapse formation. To probe the phylogenetic origins of RGK protein–Ca2+ channel interactions, we identified potential RGK-like protein homologs in genomes for genetically diverse organisms from both the deuterostome and protostome animal superphyla. RGK-like protein homologs cloned from Danio rerio (zebrafish) and Drosophila melanogaster (fruit flies) expressed in mammalian sympathetic neurons decreased Ca2+ current density as reported for expression of mammalian RGK proteins. Sequence alignments from evolutionarily diverse organisms spanning the protostome/deuterostome divide revealed conservation of residues within the RGK G-domain involved in RGK protein – Cav? subunit interaction. In addition, the C-terminal eleven residues were highly conserved and constituted a signature sequence unique to RGK proteins but of unknown function. Taken together, these data suggest that RGK proteins, and the ability to modify Ca2+ channel function, arose from an ancestor predating the protostomes split from deuterostomes approximately 550 million years ago.

Puhl, Henry L.; Lu, Van B.; Won, Yu-Jin; Sasson, Yehezkel; Hirsch, Joel A.; Ono, Fumihito; Ikeda, Stephen R.

2014-01-01

315

Cloning and characterization of a zebrafish homologue of human AQP1: a bifunctional water and gas channel.  

PubMed

The mammalian aquaporins AQP1, AQP4, and AQP5 have been shown to function not only as water channels but also as gas channels. Zebrafish have two genes encoding an AQP1 homologue, aqp1a and aqp1b. In the present study, we cloned the cDNA that encodes the zebrafish protein Aqp1a from the 72-h postfertilization (hpf) embryo of Danio rerio, as well as from the swim bladder of the adult. The deduced amino-acid sequence of aqp1a consists of 260 amino acids and is 59% identical to human AQP1. By analyzing the genomic DNA sequence, we identified four exons in the aqp1a gene. By in situ hybridization, aqp1a is expressed transiently in the developing vasculature and in erythrocytes from 16 to 48 h of development. Later, at 72 hpf, aqp1a is expressed in dermal ionocytes and in the swim bladder. Western blot analysis of adult tissues reveals that Aqp1a is most highly expressed in the eye and swim bladder. Xenopus oocytes expressing aqp1a have a channel-dependent (*) osmotic water permeability (P(f)(*)) that is indistinguishable from that of human AQP1. On the basis of the magnitude of the transient change in surface pH (?pH(S)) that were recorded as the oocytes were exposed to either CO(2) or NH(3), we conclude that zebrafish Aqp1a is permeable to both CO(2) and NH(3). The ratio (?pH(S)(*))((CO)2)/P(f)(*) is about half that of human AQP1, and the ratio (?pH(S)(*))(NH3)/P(f)(*) is about one-quarter that of human AQP1. Thus, compared with human AQP1, zebrafish Aqp1a has about twice the selectivity for CO(2) over NH(3). PMID:20739606

Chen, Li-Ming; Zhao, Jinhua; Musa-Aziz, Raif; Pelletier, Marc F; Drummond, Iain A; Boron, Walter F

2010-11-01

316

Experimental study of air-water two-phase flow through a fracture (narrow channel)  

Microsoft Academic Search

Two-phase (air-water) flow experiments were conducted in artificial horizontal fractures (narrow channels). Two experimental set-ups were utilized. One set of experiments was performed by using two glass plates (1 × 0.5 m) with a gap width of 1 mm. The second set of experiments was performed using two bricks made of baked clay (28 × 14 cm) for which three

M. Fourar; S. Bories

1995-01-01

317

Effects of Timing and Duration of Aeration on Water Quality and Production of Channel Catfish  

Microsoft Academic Search

Sccond-year (660 g) channel catfish, Ictalurus punctatus, were stocked in ponds at a density of 1,931 kg\\/ha and fed a floating, commercial diet at 74 kg\\/ha\\/day for 10 weeks. The effect of five aeration treatments on fish growth, survival, and pond water quality was evaluated. The aeration treatments were: no aeration (0 hours), aeration from 0400-0700 (3 hours), aeration from

Abdelmoez Abdalla; Robert P. Romaire

1996-01-01

318

Role of Aquaporin Water Channels in Airway Fluid Transport, Humidification, and Surface Liquid Hydration  

Microsoft Academic Search

Several aquaporin-type water channels are expressed in mammalian airways and lung: AQP1 in mi- crovascular endothelia, AQP3 in upper airway epithelia, AQP4 in upper and lower airway epithelia, and AQP5 in al- veolar epithelia. Novel quantitative methods were developed to compare airway fluid transport-related functions in wild-type mice and knockout mice deficient in these aquaporins. Lower airway humidification, measured from

Yuanlin Song; Sujatha Jayaraman; Baoxue Yang; Michael A. Matthay; A. S. Verkman

2001-01-01

319

Structure and Chromosomal Localization of a Human Water Channel (AQP3) Gene  

Microsoft Academic Search

A cDNA encoding rat AQP3, a water channel and a member of the MIP family, that is expressed predominantly in kidney medulla and colon was cloned recently. To determine the structure, tissue distribution, and chromosomal localization of the human AQP3 gene, we screened a human kidney cDNA library with rat AQP3 probe and isolated a cDNA coding for human AQP3

Kenichi Ishibashi; Sei Sasaki; Fumiko Saito; Tatsuro Ikeuchi; Fumiaki Marumo

1995-01-01

320

Antioxidative activity of protein hydrolysates prepared from alkaline-aided channel catfish protein isolates.  

PubMed

Antioxidative activity of hydrolyzed protein prepared from alkali-solubilized catfish protein isolates was studied. The isolates were hydrolyzed to 5, 15, and 30% degree of hydrolysis using the protease enzyme, Protamex. Hydrolyzed protein was separated into hydrolysates and soluble supernatants, and both of these fractions were studied for their metal chelating ability, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and their ability to inhibit the formation of thiobarbituric acid reactive substances (TBARS) in washed tilapia muscle containing tilapia hemolysate. Both hydrolysates and supernatants were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Results showed that DPPH radical scavenging ability and reducing power of catfish protein hydrolysates decreased, whereas the ORAC value, metal chelating ability, and ability to inhibit TBARS increased, with an increase in the degree of hydrolysis. Hydrolysate samples showed higher DPPH radical scavenging ability and Fe(3+) reducing ability, and supernatant samples had higher metal chelating ability. In general, low molecular weight (MW) peptides had high ORAC values and high metal chelating ability, and high MW peptides had a higher reducing power (FRAP) and were more effective in scavenging DPPH radicals. In a washed muscle model system, the ability of catfish protein hydrolysates and their corresponding supernatants to inhibit the formation of TBARS increased with an increase in the degree of hydrolysis. PMID:18662014

Theodore, Ann E; Raghavan, Sivakumar; Kristinsson, Hordur G

2008-08-27

321

AKAP proteins anchor cAMP-dependent protein kinase to KvLQT1/IsK channel complex.  

PubMed

In cardiac myocytes, the slow component of the delayed rectifier K(+) current (I(Ks)) is regulated by cAMP. Elevated cAMP increases I(Ks) amplitude, slows its deactivation kinetics, and shifts its activation curve. At the molecular level, I(Ks) channels are composed of KvLQT1/IsK complexes. In a variety of mammalian heterologous expression systems maintained at physiological temperature, we explored cAMP regulation of recombinant KvLQT1/IsK complexes. In these systems, KvLQT1/IsK complexes were totally insensitive to cAMP regulation. cAMP regulation was not restored by coexpression with the dominant negative isoform of KvLQT1 or with the cystic fibrosis transmembrane regulator. In contrast, coexpression of the neuronal A kinase anchoring protein (AKAP)79, a fragment of a cardiac AKAP (mAKAP), or cardiac AKAP15/18 restored cAMP regulation of KvLQT1/IsK complexes inasmuch as cAMP stimulation increased the I(Ks) amplitude, increased its deactivation time constant, and negatively shifted its activation curve. However, in cells expressing an AKAP, the effects of cAMP stimulation on the I(Ks) amplitude remained modest compared with those previously reported in cardiac myocytes. The effects of cAMP stimulation were fully prevented by including the Ht31 peptide (a global disruptor of protein kinase A anchoring) in the intracellular medium. We concluded that cAMP regulation of I(Ks) requires protein kinase A anchoring by AKAPs, which therefore participate with the channel protein complex underlying I(Ks). PMID:11299204

Potet, F; Scott, J D; Mohammad-Panah, R; Escande, D; Baró, I

2001-05-01

322

Channel Surface Patterning of Alternating Biomimetic Protein Combinations for Enhanced Microfluidic Tumor Cell Isolation  

PubMed Central

Here we report a new method for multi-component protein patterning in a microchannel and also a technique for improving immunoaffinity-based circulating tumor cell (CTC) capture by patterning regions of alternating adhesive proteins using the new method. The first of two proteins, anti-epithelial cell adhesion molecule (anti-EpCAM), provides the specificity for CTC capture. The second, E-selectin, increases CTC capture under shear. Patterning regions with and without E-selectin allows captured leukocytes, which also bind E-selectin and are unwanted impurities in CTC isolation, to roll a short distance and detach from the capture surface. This reduces leukocyte capture by up to 82%. The patterning is combined with a leukocyte elution step in which a calcium chelating buffer effectively deactivates E-selectin so that leukocytes may be rinsed away 60% more efficiently than with a buffer containing calcium. The alternating patterning of this biomimetic protein combination, used in conjunction with the elution step, reduce capture of leukocytes while maintaining a high tumor capture efficiency that is up to 1.9 times higher than the tumor cell capture efficiency of a surface with only anti-EpCAM. The new patterning technique described here does not require mask alignment and can be used to spatially control the immobilization of any 2 proteins or protein mixtures inside a sealed microfluidic channel.

Launiere, Cari; Gaskill, Marissa; Czaplewski, Gregory; Myung, Ja Hye; Hong, Seungpyo; Eddington, David T.

2012-01-01

323

The Dipeptidyl-Peptidase-Like Protein DPP6 Determines the Unitary Conductance of Neuronal Kv4.2 Channels  

PubMed Central

The neuronal subthreshold-operating A-type K+ current regulates electrical excitability, spike timing, and synaptic integration and plasticity. The Kv4 channels underlying this current have been implicated in epilepsy, regulation of dopamine release, and pain plasticity. However, the unitary conductance (?) of neuronal somatodendritic A-type K+ channels composed of Kv4 pore-forming subunits is larger (?7.5 pS) than that of Kv4 channels expressed singly in heterologous cells (?4 pS). Here, we examined the putative novel contribution of the dipeptidyl-peptidase-like protein-6 DPP6-S to the ? of native [cerebellar granule neuron (CGN)] and reconstituted Kv4.2 channels. Coexpression of Kv4.2 proteins with DPP6-S was sufficient to match the ? of native CGN channels; and CGN Kv4 channels from dpp6 knock-out mice yielded a ? indistinguishable from that of Kv4.2 channels expressed singly. Moreover, suggesting electrostatic interactions, charge neutralization mutations of two N-terminal acidic residues in DPP6-S eliminated the increase in ?. Therefore, DPP6-S, as a membrane protein extrinsic to the pore domain, is necessary and sufficient to explain a fundamental difference between native and recombinant Kv4 channels. These observations may help to understand the molecular basis of neurological disorders correlated with recently identified human mutations in the dpp6 gene.

De Santiago-Castillo, Jose A.; Rocha, Carmen A.; Nadal, Marcela S.; Rudy, Bernardo; Covarrubias, Manuel

2009-01-01

324

Laser-induced acoustic wave generation\\/propagation\\/interaction in water in various internal channels  

Microsoft Academic Search

Short pulsed laser-induced single acoustic wave generation, propagation, interaction within a water-filled internal channel\\u000a are experimentally and numerically studied. A large-area, short-duration, single-plane acoustic wave was generated by the\\u000a thermoelastic interaction of a homogenized nanosecond pulsed laser beam with a liquid–solid interface and propagated at the\\u000a speed of sound in water. Laser flash Schlieren photography was used to visualize the

Seung Hwan Ko; Daeho Lee; Heng Pan; Sang-Gil Ryu; Costas P. Grigoropoulos; Nick Kladias; Elias Panides; Gerald A. Domoto

2010-01-01

325

Metamorphic response of the CLIC1 chloride intracellular ion channel protein upon membrane interaction.  

PubMed

A striking feature of the CLIC (chloride intracellular channel) protein family is the ability of its members to convert between a soluble state and an integral membrane channel form. Direct evidence of the structural transition required for the CLIC protein to autonomously insert into the membrane is lacking, largely because of the challenge of probing the conformation of the membrane-bound protein. However, insights into the CLIC transmembrane form can be gained by biophysical methods such as fluorescence resonance energy transfer (FRET) spectroscopy. This approach was used to measure distances from tryptophan 35, located within the CLIC1 putative N-domain transmembrane region, to three native cysteine residues within the C-terminal domain. These distances were computed both in aqueous solution and upon the addition of membrane vesicles. The FRET distances were used as constraints for modeling of a structure for the CLIC1 integral membrane form. The data are suggestive of a large conformational unfolding occurring between the N- and C-domains of CLIC1 upon interaction with the membrane. Consistent with previous findings, the N-terminal domain of CLIC1 is likely to insert into the lipid bilayer, while the C-domain remains in solution on the extravesicular side of the membrane. PMID:20507120

Goodchild, Sophia C; Howell, Michael W; Littler, Dene R; Mandyam, Ramya A; Sale, Kenneth L; Mazzanti, Michele; Breit, Samuel N; Curmi, Paul M G; Brown, Louise J

2010-06-29

326

The vitelliform macular dystrophy protein defines a new family of chloride channels.  

PubMed

Vitelliform macular dystrophy (VMD/Best disease; MIM*153700) is an early-onset autosomal dominant disorder in which accumulation of lipofuscin-like material within and beneath the retinal pigment epithelium is associated with a progressive loss of central vision. Bestrophin, the protein product of the VMD gene, has four predicted transmembrane domains. There are multiple bestrophin homologues in the human, Drosophila, and Caenorhabditis elegans genomes, but no function has previously been ascribed to these proteins, and they show no detectable homology to other proteins of known function. Using heterologous expression, we show here that human, Drosophila, and C. elegans bestrophins form oligomeric chloride channels, and that human bestrophin is sensitive to intracellular calcium. Each of 15 missense mutations asscociated with VMD greatly reduces or abolishes the membrane current. Four of these mutant bestrophins were coexpressed with the wild type and each dominantly inhibited the wild-type membrane current, consistent with the dominant nature of the disease. These experiments establish the existence of a new chloride channel family and VMD as a channelopathy. PMID:11904445

Sun, Hui; Tsunenari, Takashi; Yau, King-Wai; Nathans, Jeremy

2002-03-19

327

Comparisons of the hydraulics of water flows in Martian outflow channels with flows of similar scale on earth  

NASA Technical Reports Server (NTRS)

The hydraulics of channelized water flows on Mars and the resulting sediment transport rates are calculated, and similar computations are performed for such terrestrial analogs as the Mississippi River and the catastrophic Lake Missoula floods that formed the Channeled Scabland in eastern Washington State. The morphologies of deep-sea channels formed by catastrophic turbidity currents are compared with the Martian channels, many similarities are pointed out, and the hydraulics of the various flows are compared. The results indicate that the velocities, discharges, bottom shear stresses, and sediment-transport capacity of water flows along the Martian channels would be comparable to those of the oceanic turbidity currents and the Lake Missoula floods. It is suggested that the submarine canyons from which turbidity currents originate are the terrestrial counterparts to the chaotic-terrain areas or craters that serve as sources for many of the Martian channels.

Komar, P. D.

1979-01-01

328

Cloud area determination from AVIRIS data using water vapor channels near 1. mu. m  

SciTech Connect

Fractional cloud area is derived from spectral images collected by the airborne visible-infrared imaging spectrometer (AVIRIS). AVIRIS covers the spectral region from 0.4 to 2.5 {mu}m with 224 channels and has a ground instantaneous field of view of 20 {times} 20 m from an altitude of 20 km. The derivation is made by ratioing radiances near the 0.94- and the 1.14-{mu}m water vapor band centers against those in the intermediate atmospheric window regions. The derivation makes use of the facts that (1) the reflectances of most ground targets vary approximately linearly with wavelength in the 0.94- and the 1.14-{mu}m water vapor band absorption regions, and (2) the peak absorptions of the water vapor band over cloudy areas are smaller than those over nearby clear surface areas because of the rapidly decreasing atmospheric water vapor concentration with height. The band ratioing technique effectively discriminates among clouds and surface areas having similar reflectance values. Such discrimination is not possible using standard radiance thresholding techniques. It is expected that the use of water vapor channels in the near-infrared region in future satellites will improve the ability to determine cloud cover over the land.

Gao, B.C.; Goetz, A.F.H. (Univ. of Colorado, Boulder (USA))

1991-02-20

329

Tissue-speci¢c regulation of Ca2á channel protein expression by sex hormones  

Microsoft Academic Search

The L-type Ca2á channel pore-forming K subunit, K1C can be detected in brain and heart as two proteins with molecular masses of V240 kDa and V190 kDa known as K1Clong and K1Cshort, respectively. In brain, the K1Cshort is thought to be the product of a V50 kDa C- terminus calpain-mediated proteolytic deletion. We now show that uterine smooth muscle also

Gustavo Helguera; Riccardo Olcese; Min Song; Ligia Toro; Enrico Stefani

330

Tissue-specific regulation of Ca 2+ channel protein expression by sex hormones  

Microsoft Academic Search

The L-type Ca2+ channel pore-forming ? subunit, ?1C can be detected in brain and heart as two proteins with molecular masses of ?240 kDa and ?190 kDa known as ?1C-long and ?1C-short, respectively. In brain, the ?1C-short is thought to be the product of a ?50 kDa C-terminus calpain-mediated proteolytic deletion. We now show that uterine smooth muscle also possesses

Gustavo Helguera; Riccardo Olcese; Min Song; Ligia Toro; Enrico Stefani

2002-01-01

331

Terahertz Dance of Proteins and Sugars with Water  

NASA Astrophysics Data System (ADS)

The influence of water on biomolecular interfaces and functionality has been in the focus of hydration studies. Improved experimental and computational probes gave insight to this question from different perspectives. The aspect of collective water network dynamics has been experimentally accessed by terahertz (THz) spectroscopy, which is sensitive to even small solute-induced rearrangements of the water network in the biomolecular surroundings. THz hydration studies uncovered that the dynamical hydration shell of saccharides consists of several hundred water molecules and up to thousand water molecules for proteins. Mutations at the protein surface and inside the core perturb the dynamical hydration, whereas it is noticeable that native wild-type proteins most significantly affect hydration dynamics. Kinetic THz absorption (KITA) studies of protein folding recently revealed that solvent dynamics are coupled to secondary structure formation of the protein. The solvent water network is dynamically rearranged in milliseconds before the protein folds to its native state within the following seconds. THz spectroscopy gives experimental evidence that collective long-range dynamics are a key factor of biomolecular hydration.

Born, Benjamin; Havenith, Martina

2009-12-01

332

Rh proteins vs Amt proteins: an organismal and phylogenetic perspective on CO2 and NH3 gas channels.  

PubMed

Rh (Rhesus) proteins are homologues of ammonium transport (Amt) proteins. Physiological and structural evidence shows that Amt proteins are gas channels for NH(3), but the substrate of Rh proteins, be it CO2 as shown in green alga, or NH3/NH4+ as shown in mammalian cells, remains disputed. We assembled a large dataset generated of Rh and Amt to explore how Rh originated from and evolved independently of Amt relatives. Analysis of this rich data implies that Rh was split from Amt first to emerge in archaeal species. The Rh ancestor underwent divergence and duplication along speciation, leading to neofunctionalization and subfunctionalization of the Rh family. The characteristic organismal distribution of Rh vs. Amt reflects their early separation and subsequent independent evolution: they coexist in microbes and invertebrates but do not in fungi, vascular plants or vertebrates. Rh gene-duplication was prominent in vertebrates: while epithelial RhBG/RhCG displayed strong purifying selection, erythroid Rh30 and RhAG experienced different episodes of positive selection in each of which adaptive evolution occurred at certain time points and in a few codon sites. Mammalian Rh30 and RhAG were subject to particularly strong positive selection in some codon sites in the lineage from rodents to human. The grounds of this adaptive evolution may be driven by the necessity to increase the surface/volume ratio of biconcave erythrocytes for facilitative gas diffusion. Altogether, these results are consistent with Rh proteins not being the orthologue of Amt proteins but having gained the function for CO2/HCO3- transport, with important roles in systemic pH regulation. PMID:16564193

Peng, J; Huang, C H

2006-01-01

333

Neuropeptide Y receptors differentially modulate G-protein-activated inwardly rectifying K+ channels and high-voltage-activated Ca2+ channels in rat thalamic neurons  

PubMed Central

Using whole-cell patch-clamp recordings, infrared videomicroscopy and fast focal solution exchange methods, the actions of neuropeptide Y (NPY) were examined in thalamic slices of postnatal (10-16 days) rats. NPY activated a K+-selective current in neurons of the thalamic reticular nucleus (RT; 20/29 neurons) and ventral basal complex (VB; 19/25 neurons). The currents in both nuclei had activation and deactivation kinetics that were very similar to those of GABAB receptor-induced currents, were totally blocked by 0.1 mM Ba2+ and showed voltage-dependent relaxation. These properties indicate that the NPY-sensitive K+ current is mediated by G-protein-activated, inwardly rectifying K+ (GIRK) channels. In RT neurons, NPY application reversibly reduced high-voltage-activated (HVA) currents to 33 ± 5 % (n= 40) of the control level but did not affect the T-type currents. Inhibition of Ca2+ currents was voltage independent and was largely mediated by effects on N- and P/Q-type channels. NPY activation of GIRK channels was mediated via NPY1 receptors, whereas inhibition of N- and P/Q-type Ca2+ channels was mediated by NPY2 receptors. These results show that neuropeptide Y activates K+ channels and simultaneously inhibits HVA Ca2+ channels via different receptor subtypes.

Sun, Qian-Quan; Huguenard, John R; Prince, David A

2001-01-01

334

Chryse Planitia region, Mars: Channeling history, flood-volume estimates, and scenarios for bodies of water in the northern plains  

NASA Technical Reports Server (NTRS)

The Chryse Planitia region of Mars includes several outflow channels that debouched into a single basin. Here we evaluate possible volumes and areal extents of standing bodies of water that collected in the northern lowland plains, based on evidence provided by topography, fluvial relations, and channel chronology and geomorphology.

Rotto, Susan L.; Tanaka, Kenneth L.

1992-01-01

335

Inspection Head Design for the In-Service Inspection of Fuel Channels of Pressurized Heavy Water Reactors  

Microsoft Academic Search

This paper discusses the challenges associated with the mechanical design of Inspection Head for the in-service inspection (ISI) of fuel channels of Indian Pressurized Heavy Water Reactors (PHWRs). ISI is carried out during shut down period in the reactor. Non Destructive Examination (NDE) of fuel channels is a mandatory requirement to acquire knowledge about the structural condition. A typical 220

Amit Kumar Haruray; R. D. Veerapur; R. K. Puri; Manjit Singh

2006-01-01

336

G-protein modulation of N-type calcium channel gating current in human embryonic kidney cells (HEK 293).  

PubMed Central

1. Voltage-dependent inhibition of N-type calcium currents by G-proteins contributes importantly to presynaptic inhibition. To examine the effect of G-proteins on key intermediary transitions leading to channel opening, we measured both gating and ionic currents arising from recombinant N-type channels (alpha 1B, beta 1b and alpha 2) expressed in transiently transfected human embryonic kidney cells (HEK 293). Recombinant expression of a homogeneous population of channels provided a favourable system for rigorous examination of the mechanisms underlying G-protein modulation. 2. During intracellular dialysis with GTP gamma S to activate G-proteins, ionic currents demonstrated classic features of voltage-dependent inhibition, i.e. strong depolarizing prepulses increased ionic currents and produced hyperpolarizing shifts in the voltage-dependent activation of ionic current. No such effects were observed with GDP beta S present to minimize G-protein activity. 3. Gating currents were clearly resolved after ionic current blockade with 0.1 mM free La3+, enabling this first report of gating charge translocation arising exclusively from N-type channels. G-proteins decreased the amplitude of gating currents and produced depolarizing shifts in the voltage-dependent activation of gating charge movement. However, the greatest effect was to induce a approximately 20 mV separation between the voltage-dependent activation of gating charge movement and ionic current. Strong depolarizing prepulses largely reversed these effects. These modulatory features provide telling clues about the kinetic steps affected by G-proteins because gating currents arise from the movement of voltage sensors that trigger channel activation. 4. The mechanistic implications of concomitant G-protein-mediated changes in gating and ionic currents are discussed. We argue that G-proteins act to inhibit both voltage-sensor movement and the transduction of voltage-sensor activation into channel opening. Images Figure 4

Jones, L P; Patil, P G; Snutch, T P; Yue, D T

1997-01-01

337

Calmodulin and S100A1 protein interact with N terminus of TRPM3 channel.  

PubMed

Transient receptor potential melastatin 3 ion channel (TRPM3) belongs to the TRP family of cation-permeable ion channels involved in many important biological functions such as pain transduction, thermosensation, and mechanoregulation. The channel was reported to play an important role in Ca(2+) homeostasis, but its gating mechanisms, functions, and regulation are still under research. Utilizing biophysical and biochemical methods, we characterized two independent domains, Ala-35-Lys-124 and His-291-Gly-382, on the TRPM3 N terminus, responsible for interactions with the Ca(2+)-binding proteins calmodulin (CaM) and S100A1. We identified several positively charged residues within these domains as having a crucial impact on CaM/S100A1 binding. The data also suggest that the interaction is calcium-dependent. We also performed competition assays, which suggested that CaM and S100A1 are able to compete for the same binding sites within the TRPM3 N terminus. This is the first time that such an interaction has been shown for TRP family members. PMID:22451665

Holakovska, Blanka; Grycova, Lenka; Jirku, Michaela; Sulc, Miroslav; Bumba, Ladislav; Teisinger, Jan

2012-05-11

338

Biogas production from water hyacinth and channel grass used for phytoremediation of industrial effluents.  

PubMed

The paper reports on the biogas production from water hyacinth (Eichhornia crassipes) and channel grass (Vallisneria spiralis) employed separately for phytoremediation of lignin and metal-rich pulp and paper mill and highly acidic distillery effluents. These plants eventually grow well in diluted effluent up to 40% (i.e., 2.5-times dilution with deionized water) and often take up metals and toxic materials from wastewater for their metabolic use. Slurry of the two plants used for phytoremediation produced significantly more biogas than that produced by the plants grown in deionized water; the effect being more marked with plants used for phytoremediation of 20% pulp and paper mill effluent. Biogas production from channel grass was relatively greater and quicker (maximum in 6-9 days) than that from water hyacinth (in 9-12 days). Such variation in biogas production by the two macrophytes has been correlated with the changes in C, N and C/N ratio of their slurry brought by phytoremediation. PMID:12688463

Singhal, V; Rai, J P N

2003-02-01

339

Population Characteristics of the Channel, Flathead, and Blue Catfish in the Indiana Illinois Interjurisdictional Waters of the Wabash River.  

National Technical Information Service (NTIS)

The objective of this project was to describe the population characteristics of the channel (Ictalurus punctatus), flathead (Pylodictis olivaris) and blue (Ictalurus furcatus) catfish in the Indiana-Illinois interjurisdictional waters of the Wabash River....

T. E. Lauer

1999-01-01

340

Hydration water dynamics and instigation of protein structuralrelaxation  

SciTech Connect

Until a critical hydration level is reached, proteins do not function. This critical level of hydration is analogous to a similar lack of protein function observed for temperatures below a dynamical temperature range of 180-220K that also is connected to the dynamics of protein surface water. Restoration of some enzymatic activity is observed in partially hydrated protein powders, sometimes corresponding to less than a single hydration layer on the protein surface, which indicates that the dynamical and structural properties of the surface water is intimately connected to protein stability and function. Many elegant studies using both experiment and simulation have contributed important information about protein hydration structure and timescales. The molecular mechanism of the solvent motion that is required to instigate the protein structural relaxation above a critical hydration level or transition temperature has yet to be determined. In this work we use experimental quasi-elastic neutron scattering (QENS) and molecular dynamics simulation to investigate hydration water dynamics near a greatly simplified protein system. We consider the hydration water dynamics near the completely deuterated N-acetyl-leucine-methylamide (NALMA) solute, a hydrophobic amino acid side chain attached to a polar blocked polypeptide backbone, as a function of concentration between 0.5M-2.0M under ambient conditions. We note that roughly 50-60% of a folded protein's surface is equally distributed between hydrophobic and hydrophilic domains, domains whose lengths are on the order of a few water diameters, that justify our study of hydration dynamics of this simple model protein system. The QENS experiment was performed at the NIST Center for Neutron Research, using the disk chopper time of flight spectrometer (DCS). In order to separate the translational and rotational components in the spectra, two sets of experiments were carried out using different incident neutron wavelengths of 7.5{angstrom} and 5.5{angstrom} to give two different time resolutions. All the spectra have been measure at room temperature. The spectra were corrected for the sample holder contribution and normalized using the vanadium standard. The resulting data were analyzed with DAVE programs (http://www.ncnr.nist.gov/dave/). The AMBER force field and SPCE water model were used for modeling the NALMA solute and water, respectively. For the analysis of the water dynamics in the NALMA aqueous solutions, we performed simulations of a dispersed solute configuration consistent with our previous structural analysis, where we had primarily focused on the structural organization of these peptide solutions and their connection to protein folding. Further details of the QENS experiment and molecular dynamics simulations are reported elsewhere.

Russo, Daniela; Hura, Greg; Head-Gordon, Teresa

2003-09-01

341

Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species  

PubMed Central

The mechanisms by which replicating influenza viruses decrease the expression and function of amiloride-sensitive epithelial sodium channels (ENaCs) have not been elucidated. We show that expression of M2, a transmembrane influenza protein, decreases ENaC membrane levels and amiloride-sensitive currents in both Xenopus oocytes, injected with human ?-, ?-, and ?-ENaCs, and human airway cells (H441 and A549), which express native ENaCs. Deletion of a 10-aa region within the M2 C terminus prevented 70% of this effect. The M2 ENaC down-regulation occurred at normal pH and was prevented by MG-132, a proteasome and lysosome inhibitor. M2 had no effect on Liddle ENaCs, which have decreased affinity for Nedd4-2. H441 and A549 cells transfected with M2 showed higher levels of reactive oxygen species, as shown by the activation of redox-sensitive dyes. Pretreatment with glutathione ester, which increases intracellular reduced thiol concentrations, or protein kinase C (PKC) inhibitors prevented the deleterious effects of M2 on ENaCs. The data suggest that M2 protein increases steady-state concentrations of reactive oxygen intermediates that simulate PKC and decrease ENaCs by enhancing endocytosis and its subsequent destruction by the proteasome. These novel findings suggest a mechanism for the influenza-induced rhinorrhea and life-threatening alveolar edema in humans.—Lazrak, A., Iles, K. E., Liu, G. Noah, D. L., Noah, J. W., Matalon, S. Influenza virus M2 protein inhibits epithelial sodium channels by increasing reactive oxygen species.

Lazrak, Ahmed; Iles, Karen E.; Liu, Gang; Noah, Diana L.; Noah, James W.; Matalon, Sadis

2009-01-01

342

Relationships Between Stream - Ground Water Exchange and Topography of the Channel, Valley, and Watershed  

NASA Astrophysics Data System (ADS)

Stream flow gains and losses represent exchange with groundwater and are commonly associated with the topography of the stream channel and contributing area. The magnitude of stream gain, i.e. runoff generation, is thought to be related to the extent and geometry of the contributing surface area. At smaller scales, the magnitude of both stream gain and loss may be related to heterogeneity in the gradient of the stream channel or valley. To validate relationships such as these between streams and their topography, we compare measurements of stream reach gains and losses to the terrain analyses of corresponding channels, valleys, and contributing areas. Comparisons are made for 26, 100-m reaches that constitute a 2.6-km long headwater stream in the Tenderfoot Creek Experimental Forest (USFS), Montana. The stream drains a 5.5 km2 catchment with a riparian area of 0.073 km2, delineated by contributing area with elevation within 2 m of the stream channel. The study stream flows over 3 geological units with valley slopes around 6.7, 5.7, and 9.0%, from upstream to downstream. For each 100-m reach, upstream and downstream discharges were measured using conservative tracer (chloride) experiments and dilution gauging techniques. In addition, the upstream release was measured at the downstream end of each reach to determine tracer mass loss and to estimate gross hydrologic loss over the reach. To close the mass balance, gross gain was calculated from the net change in discharge and gross loss. The spatial distributions of gross gains and losses were determined at multiple times during the declining summer baseflows of the snowmelt driven hydrograph. At lower baseflow conditions, several net neutral or gaining reaches also showed a 5-15% tracer mass loss, indicating that gross gain and loss operate concurrently in these reaches. We use these water balance fluxes to indicate one scale of stream - ground water exchange, and we compare patterns in exchange with the surrounding topography. Elevation data for the stream and watershed were collected using traditional survey techniques and aerial laser swath mapping (ALSM, 1-m resolution). Topographic metrics, such as channel sinuosity, valley slope, riparian area, and lateral contributing area, are calculated through terrain analyses of elevation data. Comparing topography with stream water balance is a spatially explicit approach to linking watershed structure with stream - ground water interaction, which is important to understanding solute fate and transport among the stream and adjacent ecosystems.

Payn, R. A.; Gooseff, M. N.; McGlynn, B. L.; Bencala, K. E.; Wondzell, S. M.; Jencso, K.

2007-12-01

343

AMP-activated protein kinase regulates hERG potassium channel.  

PubMed

Besides their role in cardiac repolarization, human ether-a-go-go-related gene potassium (hERG) channels are expressed in several tumor cells including rhabdomyosarcoma cells. The channels foster cell proliferation. Ubiquitously expressed AMP-dependent protein kinase (AMPK) is a serine-/threonine kinase, stimulating energy-generating and inhibiting energy-consuming processes thereby helping cells survive periods of energy depletion. AMPK has previously been shown to regulate Na?/K? ATPase, Na?/Ca²? exchangers, Ca²? channels and K? channels. The present study tested whether AMPK regulates hERG channel activity. Wild type AMPK (?1?1?1), constitutively active (?R70Q)AMPK (?1?1?1(R70Q)), or catalytically inactive (?K45R)AMPK (?1(K45R)?1?1) were expressed in Xenopus oocytes with hERG. Tail currents were determined as a measure of hERG channel activity by two-electrode-voltage clamp. hERG membrane abundance was quantified by chemiluminescence and visualized by immunocytochemistry and confocal microscopy. Moreover, hERG currents were measured in RD rhabdomyosarcoma cells after pharmacological modification of AMPK activity using the patch clamp technique. Coexpression of wild-type AMPK and of constitutively active (?R70Q)AMPK significantly downregulated the tail currents in hERG-expressing Xenopus oocytes. Pharmacological activation of AMPK with AICAR or with phenformin inhibited hERG currents in Xenopus oocytes, an effect abrogated by AMPK inhibitor compound C. (?R70Q)AMPK enhanced the Nedd4-2-dependent downregulation of hERG currents. Coexpression of constitutively active (?R70Q)AMPK decreased membrane expression of hERG in Xenopus oocytes. Compound C significantly enhanced whereas AICAR tended to inhibit hERG currents in RD rhabdomyosarcoma cells. AMPK is a powerful regulator of hERG-mediated currents in both, Xenopus oocytes and RD rhabdomyosarcoma cells. AMPK-dependent regulation of hERG may be particularly relevant in cardiac hypertrophy and tumor growth. PMID:23716168

Almilaji, Ahmad; Munoz, Carlos; Elvira, Bernat; Fajol, Abul; Pakladok, Tatsiana; Honisch, Sabina; Shumilina, Ekaterina; Lang, Florian; Föller, Michael

2013-11-01

344

The elusive character of discontinuous deep-water channels: New insights from Lucia Chica channel system, offshore California  

USGS Publications Warehouse

New high-resolution autonomous underwater vehicle (AUV) seafloor images, with 1 m lateral resolution and 0.3 m vertical resolution, reveal unexpected seafloor rugosity and low-relief (<10 m), discontinuous conduits over ~70 km2. Continuous channel thalwegs were interpreted originally from lower-resolution images, but newly acquired AUV data indicate that a single sinuous channel fed a series of discontinuous lower-relief channels. These discontinuous channels were created by at least four avulsion events. Channel relief, defined as the height from the thalweg to the levee crest, controls avulsions and overall stratigraphic architecture of the depositional area. Flowstripped turbidity currents separated into and reactivated multiple channels to create a distributary pattern and developed discontinuous trains of cyclic scours and megaflutes, which may be erosional precursors to continuous channels. The diverse features now imaged in the Lucia Chica channel system (offshore California) are likely common in modern and ancient systems with similar overall morphologies, but have not been previously mapped with lower-resolution detection methods in any of these systems. ?? 2011 Geological Society of America.

Maier, K. L.; Fildani, A.; Paull, C. K.; Graham, S. A.; McHargue, T. R.; Caress, D. W.; McGann, M.

2011-01-01

345

Recovery from Slow Inactivation in K+ Channels is Controlled by Water Molecules  

PubMed Central

The bacterial K+ channel KcsA can be used to help elucidate questions about channel inactivation and recovery at the atomic level. Although KcsA contains only a pore domain, without voltage-sensing machinery, it has the structural elements necessary for ion conduction, activation and inactivation1–7. Available X-ray structures of KcsA provide an atomic view of the four most important functional states in which the intracellular gate is either closed or open, and the selectivity filter is either conductive or inactivated8–10. Application of a specific stimulus opens the intracellular gate of a K+ channel (activation), yielding a transient period of ion conduction until the selectivity filter spontaneously undergoes a conformational change toward a non-conductive state (inactivation). Removal of the stimulus closes the gate and allows the selectivity filter to interconvert back to its conductive conformation (recovery). In this manuscript, a series of long molecular dynamics (MD) simulations reveal how the selectivity filter is sterically locked in the inactive conformation by buried water molecules bound behind the selectivity filter. Potential of mean force calculations show how the recovery process is affected by the buried waters and the rebinding of an external K+ ion. A kinetic model deduced from the simulations shows how releasing the buried waters can stretch the timescale of recovery to seconds. This leads to the prediction that reducing the occupancy of the buried waters by imposing a high osmotic stress should accelerate the rate of recovery, which was verified experimentally by measuring the recovery rate in the presence of 2M sucrose.

Ostmeyer, Jared; Chakrapani, Sudha; Pan, Albert C.; Perozo, Eduardo; Roux, Benoit

2013-01-01

346

Subunit-specific inhibition of acid sensing ion channels by stomatin-like protein 1.  

PubMed

There are five mammalian stomatin-domain genes, all of which encode peripheral membrane proteins that can modulate ion channel function. Here we examined the ability of stomatin-like protein 1 (STOML1) to modulate the proton-sensitive members of the acid-sensing ion channel (ASIC) family. STOML1 profoundly inhibits ASIC1a, but has no effect on the splice variant ASIC1b. The inactivation time constant of ASIC3 is also accelerated by STOML1. We examined STOML1 null mutant mice with a ?-galactosidase-neomycin cassette gene-trap reporter driven from the STOML1 gene locus, which indicated that STOML1 is expressed in at least 50% of dorsal root ganglion (DRG) neurones. Patch clamp recordings from mouse DRG neurones identified a trend for larger proton-gated currents in neurones lacking STOML1, which was due to a contribution of effects upon both transient and sustained currents, at different pH, a finding consistent with an endogenous inhibitory function for STOML1. PMID:24247984

Kozlenkov, Alexey; Lapatsina, Liudmila; Lewin, Gary R; Smith, Ewan St John

2014-02-15

347

IUPHAR-DB: the IUPHAR database of G protein-coupled receptors and ion channels  

PubMed Central

The IUPHAR database (IUPHAR-DB) integrates peer-reviewed pharmacological, chemical, genetic, functional and anatomical information on the 354 nonsensory G protein-coupled receptors (GPCRs), 71 ligand-gated ion channel subunits and 141 voltage-gated-like ion channel subunits encoded by the human, rat and mouse genomes. These genes represent the targets of approximately one-third of currently approved drugs and are a major focus of drug discovery and development programs in the pharmaceutical industry. IUPHAR-DB provides a comprehensive description of the genes and their functions, with information on protein structure and interactions, ligands, expression patterns, signaling mechanisms, functional assays and biologically important receptor variants (e.g. single nucleotide polymorphisms and splice variants). In addition, the phenotypes resulting from altered gene expression (e.g. in genetically altered animals or in human genetic disorders) are described. The content of the database is peer reviewed by members of the International Union of Basic and Clinical Pharmacology Committee on Receptor Nomenclature and Drug Classification (NC-IUPHAR); the data are provided through manual curation of the primary literature by a network of over 60 subcommittees of NC-IUPHAR. Links to other bioinformatics resources, such as NCBI, Uniprot, HGNC and the rat and mouse genome databases are provided. IUPHAR-DB is freely available at http://www.iuphar-db.org.

Harmar, Anthony J.; Hills, Rebecca A.; Rosser, Edward M.; Jones, Martin; Buneman, O. Peter; Dunbar, Donald R.; Greenhill, Stuart D.; Hale, Valerie A.; Sharman, Joanna L.; Bonner, Tom I.; Catterall, William A.; Davenport, Anthony P.; Delagrange, Philippe; Dollery, Colin T.; Foord, Steven M.; Gutman, George A.; Laudet, Vincent; Neubig, Richard R.; Ohlstein, Eliot H.; Olsen, Richard W.; Peters, John; Pin, Jean-Philippe; Ruffolo, Robert R.; Searls, David B.; Wright, Mathew W.; Spedding, Michael

2009-01-01

348

Novel mutations in aquaporin-2 gene in female siblings with nephrogenic diabetes insipidus: evidence of disrupted water channel function.  

PubMed

Novel mutations of the aquaporin-2 (AQP2) gene have been detected in Japanese female siblings with autosomal-recessive nephrogenic diabetes insipidus. The patients were compound heterozygote for point mutations at nucleotide position 374 (C374T) and at position 523 (G523A) in exon 2 of the AQP2 gene, resulting in substitution of methionine for threonine at codon 125 (T125M) and arginine for glycine at codon 175 (G175R). The water permeability (Pf) of oocytes injected with wild-type complementary RNA increased 9.0-fold compared with the Pf of water-injected oocytes, whereas the increases in the Pf of oocytes injected with T125M and G175R complementary RNA were only 1.7-fold and 1.5-fold, respectively. Immunoblot and immunocytochemistry indicated that the plasma membrane expressions of T125M and G175R AQP2 proteins were comparable to that of the wild-type, suggesting that although neither the T125M nor G175R mutation had a significant effect on plasma membrane expression, they both distorted the structure and function of the aqueous pore of AQP2. These results provide evidence that the nephrogenic diabetes insipidus in patients with T125M and G175R mutations is attributable not to the misrouting of AQP2, but to the disrupted water channel function. PMID:9745427

Goji, K; Kuwahara, M; Gu, Y; Matsuo, M; Marumo, F; Sasaki, S

1998-09-01

349

An aquaporin-2 water channel mutant which causes autosomal dominant nephrogenic diabetes insipidus is retained in the Golgi complex.  

PubMed Central

Mutations in the aquaporin-2 (AQP2) water channel gene cause autosomal recessive nephrogenic diabetes insipidus (NDI). Here we report the first patient with an autosomal dominant form of NDI, which is caused by a G866A transition in the AQP2 gene of one allele, resulting in a E258K substitution in the C-tail of AQP2. To define the molecular cause of NDI in this patient, AQP2-E258K was studied in Xenopus oocytes. In contrast to wild-type AQP2, AQP2-E258K conferred a small increase in water permeability, caused by a reduced expression at the plasma membrane. Coexpression of wild-type AQP2 with AQP2-E258K, but not with an AQP2 mutant in recessive NDI (AQP2-R187C), revealed a dominant-negative effect on the water permeability conferred by wild-type AQP2. The physiologically important phosphorylation of S256 by protein kinase A was not affected by the E258K mutation. Immunoblot and microscopic analyses revealed that AQP2-E258K was, in contrast to AQP2 mutants in recessive NDI, not retarded in the endoplasmic reticulum, but retained in the Golgi compartment. Since AQPs are thought to tetramerize, the retention of AQP2-E258K together with wild-type AQP2 in mixed tetramers in the Golgi compartment is a likely explanation for the dominant inheritance of NDI in this patient.

Mulders, S M; Bichet, D G; Rijss, J P; Kamsteeg, E J; Arthus, M F; Lonergan, M; Fujiwara, M; Morgan, K; Leijendekker, R; van der Sluijs, P; van Os, C H; Deen, P M

1998-01-01

350

Moving Fe2+ from ferritin ion channels to catalytic OH centers depends on conserved protein cage carboxylates.  

PubMed

Ferritin biominerals are protein-caged metabolic iron concentrates used for iron-protein cofactors and oxidant protection (Fe(2+) and O2 sequestration). Fe(2+) passage through ion channels in the protein cages, like membrane ion channels, required for ferritin biomineral synthesis, is followed by Fe(2+) substrate movement to ferritin enzyme (Fox) sites. Fe(2+) and O2 substrates are coupled via a diferric peroxo (DFP) intermediate, ?max 650 nm, which decays to [Fe(3+)-O-Fe(3+)] precursors of caged ferritin biominerals. Structural studies show multiple conformations for conserved, carboxylate residues E136 and E57, which are between ferritin ion channel exits and enzymatic sites, suggesting functional connections. Here we show that E136 and E57 are required for ferritin enzyme activity and thus are functional links between ferritin ion channels and enzymatic sites. DFP formation (Kcat and kcat/Km), DFP decay, and protein-caged hydrated ferric oxide accumulation decreased in ferritin E57A and E136A; saturation required higher Fe(2+) concentrations. Divalent cations (both ion channel and intracage binding) selectively inhibit ferritin enzyme activity (block Fe(2+) access), Mn(2+) < Co(2+) < Cu(2+) < Zn(2+), reflecting metal ion-protein binding stabilities. Fe(2+)-Cys126 binding in ferritin ion channels, observed as Cu(2+)-S-Cys126 charge-transfer bands in ferritin E130D UV-vis spectra and resistance to Cu(2+) inhibition in ferritin C126S, was unpredicted. Identifying E57 and E136 links in Fe(2+) movement from ferritin ion channels to ferritin enzyme sites completes a bucket brigade that moves external Fe(2+) into ferritin enzymatic sites. The results clarify Fe(2+) transport within ferritin and model molecular links between membrane ion channels and cytoplasmic destinations. PMID:24843174

Behera, Rabindra K; Theil, Elizabeth C

2014-06-01

351

Water and Backbone Dynamics in a Hydrated Protein  

PubMed Central

Abstract Rotational immobilization of proteins permits characterization of the internal peptide and water molecule dynamics by magnetic relaxation dispersion spectroscopy. Using different experimental approaches, we have extended measurements of the magnetic field dependence of the proton-spin-lattice-relaxation rate by one decade from 0.01 to 300 MHz for 1H and showed that the underlying dynamics driving the protein 1H spin-lattice relaxation is preserved over 4.5 decades in frequency. This extension is critical to understanding the role of 1H2O in the total proton-spin-relaxation process. The fact that the protein-proton-relaxation-dispersion profile is a power law in frequency with constant coefficient and exponent over nearly 5 decades indicates that the characteristics of the native protein structural fluctuations that cause proton nuclear spin-lattice relaxation are remarkably constant over this wide frequency and length-scale interval. Comparison of protein-proton-spin-lattice-relaxation rate constants in protein gels equilibrated with 2H2O rather than 1H2O shows that water protons make an important contribution to the total spin-lattice relaxation in the middle of this frequency range for hydrated proteins because of water molecule dynamics in the time range of tens of ns. This water contribution is with the motion of relatively rare, long-lived, and perhaps buried water molecules constrained by the confinement. The presence of water molecule reorientational dynamics in the tens of ns range that are sufficient to affect the spin-lattice relaxation driven by 1H dipole-dipole fluctuations should make the local dielectric properties in the protein frequency dependent in a regime relevant to catalytically important kinetic barriers to conformational rearrangements.

Diakova, Galina; Goddard, Yanina A.; Korb, Jean-Pierre; Bryant, Robert G.

2010-01-01

352

Developmental changes in the cochlear hair cell mechanotransducer channel and their regulation by transmembrane channel-like proteins  

PubMed Central

Vibration of the stereociliary bundles activates calcium-permeable mechanotransducer (MT) channels to initiate sound detection in cochlear hair cells. Different regions of the cochlea respond preferentially to different acoustic frequencies, with variation in the unitary conductance of the MT channels contributing to this tonotopic organization. Although the molecular identity of the MT channel remains uncertain, two members of the transmembrane channel–like family, Tmc1 and Tmc2, are crucial to hair cell mechanotransduction. We measured MT channel current amplitude and Ca2+ permeability along the cochlea’s longitudinal (tonotopic) axis during postnatal development of wild-type mice and mice lacking Tmc1 (Tmc1?/?) or Tmc2 (Tmc2?/?). In wild-type mice older than postnatal day (P) 4, MT current amplitude increased ?1.5-fold from cochlear apex to base in outer hair cells (OHCs) but showed little change in inner hair cells (IHCs), a pattern apparent in mutant mice during the first postnatal week. After P7, the OHC MT current in Tmc1?/? (dn) mice declined to zero, consistent with their deafness phenotype. In wild-type mice before P6, the relative Ca2+ permeability, PCa, of the OHC MT channel decreased from cochlear apex to base. This gradient in PCa was not apparent in IHCs and disappeared after P7 in OHCs. In Tmc1?/? mice, PCa in basal OHCs was larger than that in wild-type mice (to equal that of apical OHCs), whereas in Tmc2?/?, PCa in apical and basal OHCs and IHCs was decreased compared with that in wild-type mice. We postulate that differences in Ca2+ permeability reflect different subunit compositions of the MT channel determined by expression of Tmc1 and Tmc2, with the latter conferring higher PCa in IHCs and immature apical OHCs. Changes in PCa with maturation are consistent with a developmental decrease in abundance of Tmc2 in OHCs but not in IHCs.

Kim, Kyunghee X.

2013-01-01

353

The Nedd4-like protein KIAA0439 is a potential regulator of the epithelial sodium channel.  

PubMed

The amiloride-sensitive epithelial sodium channel (ENaC) plays a critical role in fluid and electrolyte homeostasis and consists of alpha, beta, and gamma subunits. The carboxyl terminus of each ENaC subunit contains a PPxY, motif which is believed to be important for interaction with the WW domains of the ubiquitin-protein ligase, Nedd4. Disruption of this interaction, as in Liddle's syndrome, where mutations delete or alter the PPxY motif of either the beta or gamma subunits, has been proposed to result in increased ENaC activity. Here we present evidence that KIAA0439 protein, a close relative of Nedd4, is also a potential regulator of ENaC. We demonstrate that KIAA0439 WW domains bind all three ENaC subunits. We show that a recombinant KIAA0439 WW domain protein acts as a dominant negative mutant that can interfere with the Na(+)-dependent feedback inhibition of ENaC in whole-cell patch clamp experiments. We propose that KIAA0439 and Nedd4 proteins either play a redundant role in ENaC regulation or function in a tissue- and/or signal-specific manner to down-regulate ENaC. PMID:11244092

Harvey, K F; Dinudom, A; Cook, D I; Kumar, S

2001-03-16

354

Modification of cardiac sodium channels by carboxyl reagents. Trimethyloxonium and water-soluble carbodiimide  

PubMed Central

In TTX-sensitive nerve and skeletal muscle Na+ channels, selective modification of external carboxyl groups with trimethyloxonium (TMO) or water-soluble carbodiimide (WSC) prevents voltage-dependent Ca2+ block, reduces unitary conductance, and decreases guanidinium toxin affinity. In the case of TMO, it has been suggested that all three effects result from modification of a single carboxyl group, which causes a positive shift in the channel's surface potential. We studied the effect of these reagents on Ca2+ block of adult rabbit ventricular Na+ channels in cell-attached patches. In unmodified channels, unitary conductance (gamma Na) was 18.6 +/- 0.9 pS with 280 mM Na+ and 2 mM Ca2+ in the pipette and was reduced to 5.2 +/- 0.8 pS by 10 mM Ca2+. In contrast to TTX-sensitive Na+ channels, Ca2+ block of cardiac Na+ channels was not prevented by TMO; after TMO pretreatment, gamma Na was 6.1 +/- 1.0 pS in 10 mM Ca2+. Nevertheless, TMO altered cardiac Na+ channel properties. In 2 mM Ca2+, TMO-treated patches exhibited up to three discrete gamma Na levels: 15.3 +/- 1.7, 11.3 +/- 1.5, and 9.8 +/- 1.8 pS. Patch-to-patch variation in which levels were present and the absence of transitions between levels suggests that at least two sites were modified by TMO. An abbreviation of mean open time (MOT) accompanied each decrease in gamma Na. The effects on channel gating of elevating external Ca2+ differed from those of TMO pretreatment. Increasing pipette Ca2+ from 2 to 10 mM prolonged the MOT at potentials positive to approximately -35 mV by decreasing the open to inactivated (O-->I) transition rate constant. On the other hand, even in 10 mM Ca2+ TMO accelerated the O-->I transition rate constant without a change in its voltage dependence. Ensemble averages after TMO showed a shortening of the time to peak current and an acceleration of the rate of current decay. Channel modification with WSC resulted in analogous effects to those of TMO in failing to show relief from block by 10 mM Ca2+. Further, WSC caused a decrease in gamma Na and an abbreviation of MOT at all potentials tested. We conclude that a change in surface potential caused by a single carboxyl modification is inadequate to explain the effects of TMO and WSC in heart. Failure of TMO and WSC to prevent Ca2+ block of the cardiac Na+ channel is a new distinction among isoforms in the Na+ channel multigene family.

1993-01-01

355

Estimation of water conductivity of the natural flood channels on the Tisza flood-plain, the Great Hungarian Plain  

NASA Astrophysics Data System (ADS)

The rapidly subsiding central part of the Pannonian Basin, the flood-plain system of the Tisza River, is analyzed. Natural flood-conducting channels that were functioning prior to the construction of the water control measures of the 19th century have been identified and mapped. By now these channels have mostly disappeared; only small traces of them can be found on modern maps. The identification of these channels was achieved by studying historical maps of the region and by geomorphologic studies. Drawing the outline of the channels and the estimation of their initial meander size was supported by elevation models and satellite imagery. Eight flood breakout points and five independent channel systems were identified. The flood conveying capacity of each channel was also estimated, based on meander wavelengths. The reliability of this estimation is discussed. The channels functioned as anabranches, conducting the floods of the Tisza to its tributary, the Körös River. The efficiency of the flood control system could be increased by regeneration of these natural channels. The summarized water conveying capacity of the abandoned channels is estimated as 1000 m 3/s, a value that is slightly under a half of the flood discharge of the Tisza River.

Timár, Gábor; Gábris, Gyula

2008-06-01

356

Infiltration and quality of water for two arroyo channels, Albuquerque, New Mexico, 1988-92  

USGS Publications Warehouse

Selected reaches of Grant Line Arroyo and Tijeras Arroyo in Albuquerque, New Mexico, were studied to collect information about the amount and quality of infiltration through arroyo channels. Infiltration rate was calculated for selected reaches of Grant Line Arroyo and Tijeras Arroyo based on instantaneous streamflow-loss volumes, wetted channel area, and instantaneous evaporation rates measured during 1988-92. Infiltration rates at Grant Line Arroyo ranged from 0.0 to 0.6 foot per day, and at Tijeras Arroyo from 2.28 to 30 feet per day. The evaporation rate ranged from one-tenth of 1 percent to 2 percent of the infiltration rate. Infiltration rates differed with the location of the reach isolated for measurement and with the time of day of the infiltration-rate measurement. Differences in intrinsic permeability of the sediments may be the most important factor affecting spatial variations in infiltration. The most important factor affecting temporal variations in infiltration may be the temperature of the water and sediment where infiltration occurs. Annual evaporation rates were greatest over saturated stream sediments and ranged from 802 to 1,025 millimeters per year or from 31.57 to 40.35 inches per year. Annual evaporation rates were least over unsaturated, unvegetated soil and ranged from 174 to 291 millimeters per year or from 6.85 to 11.46 inches per year. Annual evapotranspiration rates over grasses or shrubs or both were about one-half the rates over saturated stream sediments. Rates were similar for Grant Line and Tijeras Arroyos. The land- surface vegetation, availability of water at the land surface, availability of energy to enable a change of state from water to vapor, existence of a vapor concentration gradient, and a turbulent atmosphere to carry the vapor away may be the factors that determine the amount of evaporation and evapotranspiration. Water in Grant Line Arroyo and Tijeras Arroyo met U. S. Environmental Protection Agency drinking-water regulations for nitrate, volatile organic compounds, dissolved lead, and dissolved and total arsenic, barium, cadmium, chromium, copper, iron, silver, zinc, selenium, chloride, and sulfate concentrations. Total lead concentration in one sample from Tramway Floodway Channel, a tributary to Tijeras Arroyo, was 55 micrograms per liter, exceeding the Environmental Protection Agency drinking-water regulation of 50 micrograms per liter. Dissolved-solids concentrations calculated from the sum of cations and anions usually exceeded the Environmental Protection Agency drinking-water dissolved-solids regulation of 500 milligrams per liter at Tijeras Arroyo above Four Hills Bridge.

Thomas, Carole L.

1995-01-01

357

Ground-water and surface-water-level data at Rindge Tract on the Stockton Deep Water Ship Channel, San Joaquin County, California, 1983-84  

USGS Publications Warehouse

The Sacramento-San Joaquin Delta is formed at the confluence of the two major rivers that drain the Central Valley of California. The Sacramento and San Joaquin Rivers and many interconnecting sloughs meandered back and forth across the tidelands, frequently overflowing their banks. Approximately 1 ,100 miles of levees were constructed to form about 60 tracts or islands that protect these lands from periodic flooding. The levees were constructed of sand, silt, and peat dredged from the channel bottom and are subject to erosion and failure. Owing to compaction, oxidation of the peat, and other related conditions, the islands are subsiding at rates of up to 0.25 ft/yr. The altitude of the land surface of the islands is often below sea level and below the surface water level in the channel. This condition causes stresses that may contribute to high groundwater levels and levee failure. The U.S. Army Corps of Engineers requested that the U.S. Geological Survey install and maintain continuous recorders to monitor water levels in each of four wells. Monitoring which began in July 1983 also provided data to show the relation between surface water levels in the channel and groundwater levels in the wells. Dredging began in the area of the Rindge Tract site during the latter part of July 1983. Water levels in all four wells dropped 1.5 to 2 ft between September 1983 and September 1984 and continued to drop thorough December 1984. (Lantz-PTT)

Pierce, Michael J.; Johnson, Karen L.

1986-01-01

358

Water and sediment budgets for the stormwater-drainage channel at the Navy Ships Parts Control Center near Mechanicsburg, Pennsylvania, water year 1993  

USGS Publications Warehouse

The Navy Ships Parts Control Center near Mechanicsburg, Pa., occupies an area of 824 acres, of which 358 are covered by impervious surfaces. Most of the impervious area is drained by stormwater systems that discharge to an open channel that extends about 7,900 feet from its headwaters to its confluence with Trindle Spring Run. The channel drains an area of 992 acres, of which 435 are covered by impervious surfaces. The entire area of the Center including the stormwater-drainage channel is situated in karst terrain. Parts of the drainage channel contain large sinkholes and most of the storm runoff that enters the channel drains to the sinkholes. From 1992 to 1994, the U.S. Geological Survey, in cooperation with the Department of the Navy, conducted a detailed study of water and sediment flows in the stormwater-drainage channel. The purpose of this study was to quantify the discharge of stormwater and suspended sediment to the ground-water system, by way of sinkholes, and to Trindle Spring Run. From October 1, 1992, to September 30, 1993, the data-collection period for the study, discharge and suspended-sediment concentrations were measured at three sites along the drainage channel. During the period, water inflow to the channel totaled 679 acre-feet and outflow to Trindle Spring Run totaled 131 acre-feet. Water loss to sinkholes in the drainage channel totaled 548 acre-feet or 81 percent of inflow. Total sediment inflow to the drainage channel was 97 tons, outflow to Trindle Spring Run was 22 tons, sediment loss to sinkholes was 63 tons, and the residual 12 tons of sediment was deposited in the channel. The effect of filling the sinkholes on flooding was estimated through use of a step-backwater model. The model was used to simulate undampened water-surface elevations that would result from the maximum instantaneous discharge recorded during October 1992-September 1993. The model is constrained by uncertainty in the values of the channel-roughness parameter. Analysis of the model results indicates that during high flows, inflow to sinkholes results in a moderate reduction in discharge and water-surface elevations in the drainage channel. This analysis shows that filling the sinkholes will result in increased frequency and magnitude of flooding in downstream parts of the drainage channel and increased discharge of storm runoff and suspended sediment to Trindle Spring Run.

Reed, L. A.; Durlin, R. R.; Bender, J. K.

1994-01-01

359

MM-PBSA Captures Key Role of Intercalating Water Molecules at a Protein-Protein Interface  

PubMed Central

The calculation of protein interaction energetics is of fundamental interest, yet accurate quantities are difficult to obtain due to the complex and dynamic nature of protein interfaces. This is further complicated by the presence of water molecules, which can exhibit transient interactions of variable duration and strength with the protein surface. The T-cell receptor (TCR) and its staphylococcal enterotoxin 3 (SEC3) binding partner are well-characterized examples of a protein?protein interaction system exhibiting interfacial plasticity, cooperativity, and additivity among mutants. Specifically engineered mutants induce intercalating interfacial water molecules, which subsequently enhance protein?protein binding affinity. In this work, we perform a set of molecular mechanics (MM) Poisson?Boltzmann (PB) surface area (SA) calculations on the wild type and two mutant TCR-SEC3 systems and show that the method is able to discriminate between weak and strong binders only when key explicit water molecules are included in the analysis. The results presented here point to the promise of MM-PBSA toward rationalizing molecular recognition at protein?protein interfaces, while establishing a general approach to handle explicit interfacial water molecules in such calculations.

2009-01-01

360

Plumes and Blooms: Modeling the Case II Waters of the Santa Barbara Channel. Chapter 15  

NASA Technical Reports Server (NTRS)

The goal of the Plumes and Blooms (PnB) project is to develop, validate and apply to imagery state-of-the-art ocean color algorithms for quantifying sediment plumes and phytoplankton blooms for the Case II environment of the Santa Barbara Channel. We conduct monthly to twice-monthly transect observations across the Santa Barbara Channel to develop an algorithm development and product validation data set. The PnB field program started in the summer of 1996. At each of the 7 PnB stations, a complete verification bio-geo-optical data set is collected. Included are redundant measures of apparent optical properties (remote sensing reflectance and diffuse attenuation spectra), as well as in situ profiles of spectral absorption, beam attenuation and backscattering coefficients. Water samples are analyzed for component in vivo absorption spectra, fluorometric chlorophyll, phytoplankton pigment (by the SDSU CHORS laboratory), and inorganic nutrient concentrations. A primary goal is to use the PnB field data set to objectively tune semi-analytical models of ocean color for this site and apply them using available satellite imagery (SeaWiFS and MODIS). In support of this goal, we have also been addressing SeaWiFS ocean color and AVHRR SST imagery. We also are using the PnB data set to address time/space variability of water masses in the Santa Barbara Channel and its relationship to the 1997/1998 El Nino. However, the comparison between PnB field observations and satellite estimates of primary products has been disappointing. We find that field estimates of water-leaving radiance, L(sub wN)(lambda), correspond poorly to satellite estimates for both SeaWiFS and MODIS local area coverage imagery. We believe this is due to poor atmospheric correction due to complex mixtures of aerosol types found in these near-coastal regions. Last, we remain active in outreach activities.

Siegel, D. A.; Maritorena, S.; Nelson, N. B.

2003-01-01

361

Role of mixed boundaries on flow in open capillary channels with curved air-water interfaces.  

PubMed

Flow in unsaturated porous media or in engineered microfluidic systems is dominated by capillary and viscous forces. Consequently, flow regimes may differ markedly from conventional flows, reflecting strong interfacial influences on small bodies of flowing liquids. In this work, we visualized liquid transport patterns in open capillary channels with a range of opening sizes from 0.6 to 5.0 mm using laser scanning confocal microscopy combined with fluorescent latex particles (1.0 ?m) as tracers at a mean velocity of ?0.50 mm s(-1). The observed velocity profiles indicate limited mobility at the air-water interface. The application of the Stokes equation with mixed boundary conditions (i.e., no slip on the channel walls and partial slip or shear stress at the air-water interface) clearly illustrates the increasing importance of interfacial shear stress with decreasing channel size. Interfacial shear stress emerges from the velocity gradient from the adjoining no-slip walls to the center where flow is trapped in a region in which capillary forces dominate. In addition, the increased contribution of capillary forces (relative to viscous forces) to flow on the microscale leads to increased interfacial curvature, which, together with interfacial shear stress, affects the velocity distribution and flow pattern (e.g., reverse flow in the contact line region). We found that partial slip, rather than the commonly used stress-free condition, provided a more accurate description of the boundary condition at the confined air-water interface, reflecting the key role that surface/interface effects play in controlling flow behavior on the nanoscale and microscale. PMID:22867425

Zheng, Wenjuan; Wang, Lian-Ping; Or, Dani; Lazouskaya, Volha; Jin, Yan

2012-09-01

362

Energetics of the protein-DNA-water interaction  

PubMed Central

Background To understand the energetics of the interaction between protein and DNA we analyzed 39 crystallographically characterized complexes with the HINT (Hydropathic INTeractions) computational model. HINT is an empirical free energy force field based on solvent partitioning of small molecules between water and 1-octanol. Our previous studies on protein-ligand complexes demonstrated that free energy predictions were significantly improved by taking into account the energetic contribution of water molecules that form at least one hydrogen bond with each interacting species. Results An initial correlation between the calculated HINT scores and the experimentally determined binding free energies in the protein-DNA system exhibited a relatively poor r2 of 0.21 and standard error of ± 1.71 kcal mol-1. However, the inclusion of 261 waters that bridge protein and DNA improved the HINT score-free energy correlation to an r2 of 0.56 and standard error of ± 1.28 kcal mol-1. Analysis of the water role and energy contributions indicate that 46% of the bridging waters act as linkers between amino acids and nucleotide bases at the protein-DNA interface, while the remaining 54% are largely involved in screening unfavorable electrostatic contacts. Conclusion This study quantifies the key energetic role of bridging waters in protein-DNA associations. In addition, the relevant role of hydrophobic interactions and entropy in driving protein-DNA association is indicated by analyses of interaction character showing that, together, the favorable polar and unfavorable polar/hydrophobic-polar interactions (i.e., desolvation) mostly cancel.

Spyrakis, Francesca; Cozzini, Pietro; Bertoli, Chiara; Marabotti, Anna; Kellogg, Glen E; Mozzarelli, Andrea

2007-01-01

363

Potent Neutralization of Influenza A Virus by a Single-Domain Antibody Blocking M2 Ion Channel Protein  

Microsoft Academic Search

Influenza A virus poses serious health threat to humans. Neutralizing antibodies against the highly conserved M2 ion channel is thought to offer broad protection against influenza A viruses. Here, we screened synthetic Camel single-domain antibody (VHH) libraries against native M2 ion channel protein. One of the isolated VHHs, M2-7A, specifically bound to M2-expressed cell membrane as well as influenza A

Guowei Wei; Weixu Meng; Haijiang Guo; Weiqi Pan; Jinsong Liu; Tao Peng; Ling Chen; Chang-You Chen

2011-01-01

364

Cloning and functional analysis of tipE, a novel membrane protein that enhances drosophila para sodium channel function  

Microsoft Academic Search

Voltage-dependent sodium channels are involved in the initiation and propagation of action potentials in many excitable cells. Here we report that tipE, a gene defined by a temperature-sensitive paralytic mutation in Drosophila, encodes a novel integral membrane protein that dramatically stimulates functional expression in Xenopus oocytes of the Drosophila sodium channel ? subunit encoded by the paralytic (para) locus. Using

Guoping Feng; Péter Deak; Maninder Chopra; Linda M Hall

1995-01-01

365

Role of phosphoinositol 4,5-bisphosphate and diacylglycerol in regulating native TRPC channel proteins in vascular smooth muscle  

Microsoft Academic Search

Stimulation of receptor-operated (ROCs) and store-operated (SOCs) Ca2+-permeable cation channels by vasoconstrictors has many important physiological functions in vascular smooth muscle. The present review indicates that ROCs and SOCs with diverse properties in different blood vessels are likely to be explained by composition of different subunits from the canonical transient receptor potential (TRPC) family of cation channel proteins. In addition

William A. Large; Sohag N. Saleh; Anthony P. Albert

2009-01-01

366

Maxi K + channels are stimulated by cyclic guanosine monophosphate-dependent protein kinase in canine coronary artery smooth muscle cells  

Microsoft Academic Search

By using a patch clamp technique, we examined the effect of cyclic guanosine monophosphate (cGMP)-dependent protein kinase (G kinase) on Ca2+-activated maxi K+ channels in canine coronary artery smooth muscle cells. Maxi K+ channels (274±4 pS in symmetrical 140 mM KCl at 24–26°C) were activated by cytoplasmic Ca2+ and were completely blocked by 100 nM charybdotoxin (CTX). G kinase (300

Junichi Taniguchi; Ken-Ichi Furukawa; Munekazu Shigekawa

1993-01-01

367

Voltage-Dependent Anion Channel Proteins in Synaptosomes of the Torpedo Electric Organ: Immunolocalization, Purification, and Characterization  

Microsoft Academic Search

In this study, we purified and characterized the voltage-dependent anion channel (VDAC) from the Torpedo electric organ. Using immunogold labeling, VDAC was colocalized with the voltage-gated Ca2+ channel in the synaptic plasma membrane. By immunoblot analysis, five protein bands in synaptosomes isolated from the Torpedo electric organ cross reacted with two monoclonal anti-VDAC antibody. No more than about 7 to

Iris Shafir; Wei Feng; Varda Shoshan-Barmataz

1998-01-01

368

Enhanced water and cryoprotectant permeability of porcine oocytes after artificial expression of human and zebrafish aquaporin-3 channels.  

PubMed

One of the major obstacles for the vitrification of mature porcine oocytes with ethylene glycol is their low permeability to this cryoprotectant, which results in osmotic stress-induced cell damage and low survival. Pig blastocysts, on the other hand, show enhanced water and cryoprotectant permeability, which has been related to the transcriptional activation of aquaporin-3 (AQP3) channels at this stage of development. In this study, we asked if expression of cRNAs encoding two aquaglyceroporins, human AQP3 (hAQP3) or the zebrafish Aqp3b-T85A mutant, in porcine oocytes can increase their permeability. Microinjection of germinal-vesicle-stage oocytes with enhanced green fluorescent protein (EGFP) or AQP3 cRNAs resulted in the expression of the corresponding proteins in ?26% of the metaphase-II stage oocytes at 40-44?hr of in vitro culture; co-injection of EGFP cRNA appeared to be a suitable marker for oocyte selection since all EGFP-positive oocytes also expressed the corresponding aquaporin. Using this method, we found that mature oocytes co-expressing EGFP and hAQP3 or EGFP and Aqp3b-T85A showed approximately a twofold increase of the hydraulic conductivity (Lp ) with respect non-injected or EGFP alone-injected oocytes in a 0.43?M sucrose or 1.3?M ethylene glycol solution, whereas the ethylene glycol permeability (PEG ) of EGFP?+?hAQP3 and EGFP?+?Aqp3b-T85A oocytes was 6.7- and 12-fold higher, respectively, than control oocytes. These data demonstrate that the artificial expression of aquaglyceroporins in porcine metaphase-II oocytes improves their permeability, and that the zebrafish Aqp3b-T85A mutant is more efficient than the human channel at increasing the oocyte permeability to ethylene glycol. Mol. Reprod. Dev. 81: ???-???, 2014. © 2014 Wiley Periodicals, Inc. PMID:24488947

Morató, Roser; Chauvigné, François; Novo, Sergi; Bonet, Sergi; Cerdà, Joan

2014-05-01

369

Hydration water and bulk water in proteins have distinct properties in radial distributions calculated from 105 atomic resolution crystal structures  

PubMed Central

Water plays a critical role in the structure and function of proteins, although the experimental properties of water around protein structures are not well understood. The water can be classified by the separation from the protein surface into bulk water and hydration water. Hydration water interacts closely with the protein and contributes to protein folding, stability and dynamics, as well as interacting with the bulk water. Water potential functions are often parameterized to fit bulk water properties because of the limited experimental data for hydration water. Therefore, the structural and energetic properties of the hydration water were assessed for 105 atomic resolution (?1.0 Å) protein crystal structures with a high level of hydration water by calculating the experimental water-protein radial distribution function or surface distribution function (SDF) and water radial distribution function (RDF). Two maxima are observed in SDF: the first maximum at a radius of 2.75 Å reflects first shell and hydrogen bond interactions between protein and water, the second maximum at 3.65 Å reflects second shell and van der Waals interactions between water and non-polar atoms of protein forming clathrate-hydrate-like structures. Thus, the two shells do not overlap. The RDF showed the features of liquid water rather than solid ice. The first and second maxima of RDF at 2.75 and 4.5 Å, respectively, are the same as for bulk water, but the peaks are sharper indicating hydration water is more stable than bulk water. Both distribution functions are inversely correlated with the distribution of B factors (atomic thermal factors) for the waters, suggesting that the maxima reflect stable positions. Therefore, the average water structure near the protein surface has experimentally observable differences from bulk water. This analysis will help improve the accuracy for models of water on the protein surface by providing rigorous data for the effects of the apparent chemical potential of the water near a protein surface.

Chen, Xianfeng; Weber, Irene; Harrison, Robert W.

2009-01-01

370

Interactions with PDZ Proteins Are Required for L-Type Calcium Channels to Activate cAMP Response Element Binding Protein-Dependent Gene Expression  

Microsoft Academic Search

After brief periods of heightened stimulation, calcium entry through L-type calcium channels leads to activation of the transcription factor cAMP response element-binding protein (CREB) and CRE-dependent transcription. Many of the details surrounding the mecha- nism by which L-type calcium channels are privileged in signaling to CREB, to the exclusion of other calcium entry pathways, has remained unclear. We hypothesized that

Jason P. Weick; Rachel D. Groth; Ann L. Isaksen; Paul G. Mermelstein

2003-01-01

371

Negative regulation of opioid receptor-G protein-Ca2+ channel pathway by the nootropic nefiracetam.  

PubMed

It has recently been reported that nefiracetam, a nootropic agent, is capable of attenuating the development of morphine dependence and tolerance in mice. The mechanism of this antimorphine action is not clear. The present study was designed to address this issue using Xenopus oocytes expressing delta-opioid receptors, G proteins (G(i3alpha) or G(o1alpha)), and N-type (alpha1B) Ca2+ channels. Membrane currents through Ca2+ channels were recorded from the oocytes under voltage-clamp conditions. The Ca2+ channel currents were reduced reversibly by 40-60% in the presence of 1 microM leucine-enkephalin (Leu-Enk). The Leu-Enk-induced current inhibition was recovered promptly by nefiracetam (1 microM), while control currents in the absence of Leu-Enk were not influenced by nefiracetam. A binding assay revealed that 3H-nefiracetam preferentially bound to the membrane fraction of oocytes expressing G(i3alpha). When delta-opioid receptors were coexpressed, the binding was significantly increased. However, an additional expression of alpha1B Ca2+ channels decreased the binding. The results suggest that nefiracetam preferentially binds to G(i3alpha) associated with delta-opioid receptors, thereby inhibiting the association of G proteins with Ca2+ channels. In conclusion, nefiracetam negatively regulates the inhibitory pathway of opioid receptor-G protein-Ca2+ channel. PMID:15542741

Yoshii, Mitsunobu; Furukawa, Taiji; Ogihara, Yoshiyasu; Watabe, Shigeo; Shiotani, Tadashi; Ishikawa, Yasuro; Nishimura, Masao; Nukada, Toshihide

2004-10-01

372

Convergent regulation of skeletal muscle Ca2+ channels by dystrophin, the actin cytoskeleton, and cAMP-dependent protein kinase  

PubMed Central

The skeletal muscle L-type Ca2+ channel (CaV1.1), which is responsible for initiating muscle contraction, is regulated by phosphorylation by cAMP-dependent protein kinase (PKA) in a voltage-dependent manner that requires direct physical association between the channel and the kinase mediated through A-kinase anchoring proteins (AKAPs). The role of the actin cytoskeleton in channel regulation was investigated in skeletal myocytes cultured from wild-type mice, mdx mice that lack the cytoskeletal linkage protein dystrophin, and a skeletal muscle cell line, 129 CB3. Voltage dependence of channel activation was shifted positively, and potentiation was greatly diminished in mdx myocytes and in 129 CB3 cells treated with the microfilament stabilizer phalloidin. Voltage-dependent potentiation by strong depolarizing prepulses was reduced in mdx myocytes but could be restored by positively shifting the stimulus potentials to compensate for the positive shift in the voltage dependence of gating. Inclusion of PKA in the pipette caused a negative shift in the voltage dependence of activation and restored voltage-dependent potentiation in mdx myocytes. These results show that skeletal muscle Ca2+ channel activity and voltage-dependent potentiation are controlled by PKA and microfilaments in a convergent manner. Regulation of Ca2+ channel activity by hormones and neurotransmitters that use the PKA signal transduction pathway may interact in a critical way with the cytoskeleton and may be impaired by deletion of dystrophin, contributing to abnormal regulation of intracellular calcium concentrations in dystrophic muscle.

Johnson, Barry D.; Scheuer, Todd; Catterall, William A.

2005-01-01

373

Automatic Measurement of Water Levels by Using Image Identification Method in Open Channel  

NASA Astrophysics Data System (ADS)

Water level data is indispensable to hydrology research, and it is important information for hydraulic engineering and overall utilization of water resources. The information of water level can be transmitted to management office by the network so that the management office may well understand whether the river level is exceeding the warning line. The existing water level measurement method can only present water levels in a form of data without any of images, the methods which make data just be a data and lack the sense of reality. Those images such as the rising or overflow of river level that the existing measurement method cannot obtain simultaneously. Therefore, this research employs a newly, improved method for water level measurement. Through the Video Surveillance System to record the images on site, an image of water surface will be snapped, and then the snapped image will be pre-processed and be compared with its altitude reference value to obtain a water level altitude value. With the ever-growing technology, the application scope of image identification is widely in increase. This research attempts to use image identification technology to analyze water level automatically. The image observation method used in this research is one of non-contact water level gage but it is quite different from other ones; the image observation method is cheap and the facilities can be set up beside an embankment of river or near the houses, thus the impact coming from external factors will be significantly reduced, and a real scene picture will be transmitted through wireless transmission. According to the dynamic water flow test held in an indoor experimental channel, the results of the research indicated that all of error levels of water level identification were less than 2% which meant the image identification could achieve identification result at different water levels. This new measurement method can offer instant river level figures and on-site video so that a disaster prevention measures can be made accordingly. Keywords: Image identification; Water Level; Video surveillance system.

Chung Yang, Han; Xue Yang, Jia

2014-05-01

374

Direct membrane insertion of voltage-dependent anion-selective channel protein catalyzed by mitochondrial Tom20.  

PubMed

Insertion of newly synthesized proteins into or across the mitochondrial outer membrane is initiated by import receptors at the surface of the organelle. Typically, this interaction directs the precursor protein into a preprotein translocation pore, comprised of Tom40. Here, we show that a prominent beta-barrel channel protein spanning the outer membrane, human voltage- dependent anion-selective channel (VDAC), bypasses the requirement for the Tom40 translocation pore during biogenesis. Insertion of VDAC into the outer membrane is unaffected by plugging the translocation pore with a partially translocated matrix preprotein, and mitochondria containing a temperature-sensitive mutant of Tom40 insert VDAC at the nonpermissive temperature. Synthetic liposomes harboring the cytosolic domain of the human import receptor Tom20 efficiently insert newly synthesized VDAC, resulting in transbilayer transport of ATP. Therefore, Tom20 transforms newly synthesized cytosolic VDAC into a transmembrane channel that is fully integrated into the lipid bilayer. PMID:10352015

Schleiff, E; Silvius, J R; Shore, G C

1999-05-31

375

Combinatorial expression of TRPV channel proteins defines their sensory functions and subcellular localization in C. elegans neurons.  

PubMed

C. elegans OSM-9 is a TRPV channel protein involved in sensory transduction and adaptation. Here, we show that distinct sensory functions arise from different combinations of OSM-9 and related OCR TRPV proteins. Both OSM-9 and OCR-2 are essential for several forms of sensory transduction, including olfaction, osmosensation, mechanosensation, and chemosensation. In neurons that express both OSM-9 and OCR-2, tagged OCR-2 and OSM-9 proteins reside in sensory cilia and promote each other's localization to cilia. In neurons that express only OSM-9, tagged OSM-9 protein resides in the cell body and acts in sensory adaptation rather than sensory transduction. Thus, alternative combinations of TRPV proteins may direct different functions in distinct subcellular locations. Animals expressing the mammalian TRPV1 (VR1) channel in ASH nociceptor neurons avoid the TRPV1 ligand capsaicin, allowing selective, drug-inducible activation of a specific behavior. PMID:12160748

Tobin, David; Madsen, David; Kahn-Kirby, Amanda; Peckol, Erin; Moulder, Gary; Barstead, Robert; Maricq, Andres; Bargmann, Cornelia

2002-07-18

376

Long-range protein-water dynamics in hyperactive insect antifreeze proteins.  

PubMed

Antifreeze proteins (AFPs) are specific proteins that are able to lower the freezing point of aqueous solutions relative to the melting point. Hyperactive AFPs, identified in insects, have an especially high ability to depress the freezing point by far exceeding the abilities of other AFPs. In previous studies, we postulated that the activity of AFPs can be attributed to two distinct molecular mechanisms: (i) short-range direct interaction of the protein surface with the growing ice face and (ii) long-range interaction by protein-induced water dynamics extending up to 20 Å from the protein surface. In the present paper, we combine terahertz spectroscopy and molecular simulations to prove that long-range protein-water interactions make essential contributions to the high antifreeze activity of insect AFPs from the beetle Dendroides canadensis. We also support our hypothesis by studying the effect of the addition of the osmolyte sodium citrate. PMID:23277543

Meister, Konrad; Ebbinghaus, Simon; Xu, Yao; Duman, John G; DeVries, Arthur; Gruebele, Martin; Leitner, David M; Havenith, Martina

2013-01-29

377

Sig1R Protein Regulates hERG Channel Expression through a Post-translational Mechanism in Leukemic Cells*  

PubMed Central

Sig1R (Sigma-1receptor) is a 25-kDa protein structurally unrelated to other mammalian proteins. Sig1R is present in brain, liver, and heart and is overexpressed in cancer cells. Studies using exogenous sigma ligands have shown that Sig1R interacts with a variety of ion channels, but its intrinsic function and mechanism of action remain unclear. The human ether-à-gogo related gene (hERG) encodes a cardiac channel that is also abnormally expressed in many primary human cancers, potentiating tumor progression through the modulation of extracellular matrix adhesive interactions. We show herein that sigma ligands inhibit hERG current density and cell adhesion to fibronectin in K562 myeloid leukemia cells. Heterologous expression in Xenopus oocytes demonstrates that Sig1R potentiates hERG current by stimulating channel subunit biosynthesis. Silencing Sig1R in leukemic K562 cells depresses hERG current density and cell adhesion to fibronectin by reducing hERG membrane expression. In K562 cells, Sig1R silencing does not modify hERG mRNA contents but reduces hERG mature form densities. In HEK cells expressing hERG and Sig1R, both proteins co-immunoprecipitate, demonstrating a physical association. Finally, Sig1R expression enhances both channel protein maturation and stability. Altogether, these results demonstrate for the first time that Sig1R controls ion channel expression through the regulation of subunit trafficking activity.

Crottes, David; Martial, Sonia; Rapetti-Mauss, Raphael; Pisani, Didier F.; Loriol, Celine; Pellissier, Bernard; Martin, Patrick; Chevet, Eric; Borgese, Franck; Soriani, Olivier

2011-01-01

378

Rv1698 of Mycobacterium tuberculosis Represents a New Class of Channel-forming Outer Membrane Proteins*S?  

PubMed Central

Mycobacteria contain an outer membrane composed of mycolic acids and a large variety of other lipids. Its protective function is an essential virulence factor of Mycobacterium tuberculosis. Only OmpA, which has numerous homologs in Gram-negative bacteria, is known to form channels in the outer membrane of M. tuberculosis so far. Rv1698 was predicted to be an outer membrane protein of unknown function. Expression of rv1698 restored the sensitivity to ampicillin and chloramphenicol of a Mycobacterium smegmatis mutant lacking the main porin MspA. Uptake experiments showed that Rv1698 partially complemented the permeability defect of the M. smegmatis porin mutant for glucose. These results indicated that Rv1698 provides an unspecific pore that can partially substitute for MspA. Lipid bilayer experiments demonstrated that purified Rv1698 is an integral membrane protein that indeed produces channels. The main single channel conductance is 4.5 ± 0.3 nanosiemens in 1 m KCl. Zero current potential measurements revealed a weak preference for cations. Whole cell digestion of recombinant M. smegmatis with proteinase K showed that Rv1698 is surface-accessible. Taken together, these experiments demonstrated that Rv1698 is a channel protein that is likely involved in transport processes across the outer membrane of M. tuberculosis. Rv1698 has single homologs of unknown functions in Corynebacterineae and thus represents the first member of a new class of channel proteins specific for mycolic acid-containing outer membranes.

Siroy, Axel; Mailaender, Claudia; Harder, Daniel; Koerber, Stephanie; Wolschendorf, Frank; Danilchanka, Olga; Wang, Ying; Heinz, Christian; Niederweis, Michael

2008-01-01

379

Nutritional availability of spray dried lard water protein.  

PubMed

Four growth trials were conducted with rats and chicks to investigate the nutritional value of Spray Dried Lard Water (SDLW--a by-product from the rendering process of animal lard) as a source of protein in a semipurified diet. Wean