Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.6.1.3 (ATPase)
65,361 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Surmises of how myosin subfragment 1 (S1) interacts with actin filaments in muscle contraction rest upon knowing the relative arrangement of the two proteins. Although there exist crystallographic structures for both S1 and actin, as well as electron microscopy data for the acto-S1 complex (AS1), modeling of this arrangement has so far only been done "by eye." Here we report fitted AS1 structures obtained using a quantitative method that is both more objective and makes more complete use of the data. Using undistorted crystallographic results, the best-fit AS1 structure shows significant differences from that obtained by visual fitting. The best fit is produced using the F-actin model of Holmes et al. [Holmes, K. C., Popp, D., Gebhard, W. & Kabsch, W. (1990) Nature (London) 347, 44-49]. S1 residues at the AS1 interface are now found at a higher radius as well as being translated axially and rotated azimuthally. Fits using S1 plus loops missing from the crystal structure were achieved using a homology search method to predict loop structures. These improved fits favor an arrangement in which the loop at the 50- to 20-kDa domain junction of S1 is located near the N terminus of actin. Rigid-body movements of the lower 50-kDa domain, which further improve the fit, produce closure of the large 50-kDa domain cleft and bring conserved residues in the lower 50-kDa domain into an apparently appropriate orientation for close interaction with actin. This finding supports the idea that binding of ATP to AS1 at the end of the ATPase cycle disrupts the actin binding site by changing the conformation of the 50-kDa cleft of S1.
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PMID:The structure of the acto-myosin subfragment 1 complex: results of searches using data from electron microscopy and x-ray crystallography. 923 11

Ectoenzymic activities capable of hydrolyzing ATP sequentially to adenosine are present on equine epidydimal spermatozoa membranes. Kinetic parameters for ATPase, ADPase and 5'-nucleotidase were obtained by analysis of progress reactions curve when ATP, ADP and AMP were supplied as initial substrates. These values are not different from those found when the substrates were supplied from the preceding reactions. Feed-forward inhibition on 5'-nucleotidase by ATP/ADP was taken into account to fit simulated data to the experimental results. None of the substrates supplied by the preceding reactions showed a preferential delivery to ADPase and/or 5'-nucleotidase. We therefore conclude that the model that fits the equine spermatozoa is that already proposed for pig aortic endothelial cells.
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PMID:Hydrolysis of extracellular adenine nucleotides by equine epidydimal spermatozoa. 929 97

A competition assay of 86Rb+ uptake in HeLa cells transfected with ouabain-resistant Na(+)-K(+)-ATPase mutants revealed a stimulation of 86Rb+ uptake at low external concentrations (1 mM) of competitor (K+). Of the models that were tested, those that require that two K+ be bound before transport occurs gave the worst fits. Random and ordered binding schemes described the data equally well. General models in which both binding and transport were allowed to be cooperative yielded parameter errors larger than the parameters themselves and could not be utilized. Models that assumed noncooperative transport always showed positive cooperativity in binding. E327Q and E327L mutated forms of rat alpha 2 had lower apparent affinities for the first K+ bound than did wild-type rat alpha 2 modified to be ouabain resistant. The mutations did not affect the apparent affinity of the second K+ bound. Models that assumed noncooperativity in binding always showed positively cooperative transport, i.e., enzymes with two K+ bound had a higher flux than those with one K+ bound. Increases in external Na+ decreased the apparent affinity for K+ for all models and decreased the ratio of the apparent influx rate constants for E327L.
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PMID:Mutational analysis of Glu-327 of Na(+)-K(+)-ATPase reveals stimulation of 86Rb+ uptake by external K+. 943 14

1. In rat hepatocytes under hypertonic stress, the entry of Na+ (which is thereafter exchanged for K+ via Na(+)-K(+)-ATPase) plays the key role in regulatory volume increase (RVI). 2. In the present study, the contributions of Na+ conductance, Na(+)-H+ exchange and Na(+)-K(+)-2Cl- symport to this process were quantified in confluent primary cultures by means of intracellular microelectrodes and cable analysis, microfluorometric determinations of cell pH and buffer capacity, and measurements of frusemide (furosemide)/bumetanide-sensitive 86Rb+ uptake, respectively. Osmolarity was increased from 300 to 400 mosmol l-1 by addition of sucrose. 3. The experiments indicate a relative contribution of approximately 4:1:1 to hypertonicity-induced Na+ entry for the above-mentioned transporters and the overall Na+ yield equalled 51 mmol l-1 (10 min)-1. 4. This Na+ gain is in good agreement with the stimulation of Na+ extrusion via Na(+)-K(+)-ATPase plus the actual increase in cell Na+, namely 55 mmol l-1 (10 min)-1, as we determined on the basis of ouabain-sensitive 86Rb+ uptake and by means of Na(+)-sensitive microelectrodes, respectively. 5. The overall increase in Na+ and K+ activity plus the expected concomitant increase in cell Cl- equalled 68 mmol l-1, which fits well with the increase in osmotic activity expected to occur from an initial cell shrinkage to 87.5% and a RVI to 92.6% of control, namely 53 mosmol l-1. 6. The prominent role of Na+ conductance in the RVI of rat hepatocytes could be confirmed on the basis of the pharmacological profile of this process, which was characterized by means of confocal laser-scanning microscopy.
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PMID:Role of Na+ conductance, Na(+)-H+ exchange, and Na(+)-K(+)-2Cl- symport in the regulatory volume increase of rat hepatocytes. 948 77

This work provides evidence for interactions between fragments of "19-kDa membranes," a trypsinized preparation of Na,K-ATPase that retains cation occlusion and ouabain binding. Previously, we reported rapid thermal inactivation of Rb+ occlusion at 37 degreesC (Or, E., David, P., Shainskaya, A., Tal, D. M., and Karlish, S. J. D. (1993) J. Biol. Chem. 268, 16929-16937). We describe here the detailed kinetics of thermal inactivation. In the range 25-35 degreesC, a two-step model (N left and right arrow U --> I, where N is the native species, U is the reversibly unfolded intermediate, and I is the irreversibly denatured form) fits the data. Reversibility of inactivation has been observed at 25 degreesC, consistent with the model. At 37 degreesC and higher temperatures, the data can be fitted to the simple mechanism N --> I, i.e. U is not significant as an intermediate. Occluded cations (Na+, Rb+, K+, Tl+, NH4+, and Cs+) and ouabain protect strongly against thermal inactivation. Ca2+, Ba2+, and La3+ ions do not protect. Proteolysis experiments provide independent evidence that disorganization can occur in stages, first in transmembrane segments and then in extra-membrane segments of the fragments. Analysis of selective dissociation of the M5/M6 fragment at 37 degreesC (Lutsenko, S., and Kaplan, J. H. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 7936-7940), using a specific antibody, showed that inactivation of Rb+ occlusion precedes dissociation of the fragment, and only approximately 50% of the fragment is released when occlusion is fully inactivated. In the presence of Ca2+ ions, occlusion is inactivated, but the M5/M6 fragment is not released. The experiments demonstrate that occlusion is inactivated by disruption of interactions between fragments of 19-kDa membranes, and only then does the M5/M6 fragment dissociate. Interactions between the M5/M6 and M7/M10 fragments seem to be essential for maintenance of Rb+ occlusion.
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PMID:Interactions between fragments of trypsinized Na,K-ATPase detected by thermal inactivation of Rb+ occlusion and dissociation of the M5/M6 fragment. 951 25

When unicellular algal cells are placed under anaerobic conditions, a large electrochemical gradient is built in darkness across the thylakoid membranes. We have estimated, in vivo, the amplitude of the Delta pH component of this transmembrane potential and shown that the Delta pH is twice as large as the Delta Psi. The amplitude of the Delta mu tildeH+ (approximately 110-140 mV) fits well with estimations based on the ATP/ADP ratio measured in green algae under the same conditions, suggesting that an equilibrium state is established across the thylakoid membrane. Therefore, under anaerobic dark incubation of algae, the electrochemical transmembrane potential is determined only by the cellular ATP content. The existence of this Delta mu tildeH+ is expected to result in a constitutive amount of activated CFo-CF1 ATPase, thereby facilitating ATP synthesis under low light intensity illumination. We report also on the effects of this dark-existing electrochemical gradient on the cytochrome b6f complex turnover kinetics. We show that they are largely slowed by the presence of this electrochemical transmembrane potential. The pH component is mainly responsible for the kinetic slowing down of cytochrome b6f complex turnover, despite the fact that electrogenicity is associated with the reactions taking place within this complex. Therefore, in vivo, owing to the low lumenal pH, the oxidation of plastoquinol at the Qo site is limiting the turnover of the cytochrome b6f complex in the presence of the Delta pH, while in its absence the oxidation rate of the b6 hemes becomes rate-limiting.
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PMID:In vivo characterization of the electrochemical proton gradient generated in darkness in green algae and its kinetic effects on cytochrome b6f turnover. 966 5

The human lymphoid cell activation antigen CD39 is a known E-type apyrase that hydrolyzes extracellular ATP and ADP, a function important in homotypic adhesion, platelet aggregation, and removal by activated lymphocytes of the lytic effect of ATP. The recently identified putative rat homologue of CD39L1 has been shown to have E-type ecto-ATPase activity, by hydrolyzing extracellular ATP. We have characterized three novel CD39-like transcripts, CD39L2, CD39L3, and CD39L4, which share extensive amino acid homology with other nucleotide triphosphatases in vertebrates, invertebrates, and plants, suggesting that these genes also encode proteins with ecto-nucleotidase activity. Isolation and sequencing of full-length cDNA clones for each gene identified putative proteins of 485, 529, and 429 amino acids. The expression pattern of all five human members of the gene family was analyzed. CD39L2, CD39L3, and CD39L4 were mapped on the human genome, and the murine homologues identified with the putative map locations were assigned on the basis of regions of conserved gene order between human and mouse chromosomes. The map location of mcd39l4 places the gene within a region associated with audiogenic seizure susceptibility in mouse. This disorder is characterized by convulsions induced by loud high-frequency sound and has been shown to be associated with increased nucleotide triphosphatase activity.
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PMID:The CD39-like gene family: identification of three new human members (CD39L2, CD39L3, and CD39L4), their murine homologues, and a member of the gene family from Drosophila melanogaster. 967 30

Among the major cardenolides from the milkweed Asclepias asperula, 6'-O-(E-4-hydroxycinnamoyl) desglucouzarin has not been characterized biochemically. In this study, its binding affinity for a physiological receptor, porcine kidney Na+,K+-ATPase, was found to be lower than the other cardenolides in this plant. The order of affinities from highest to lowest was: uzarigenin (Kd = 1.05 microM) = desglucouzarin (Kd = 0.98 microM) > uzarin (Kd = 4.0 microM) > 6'-O-(E-4-hydroxycinnamoyl) desglucouzarin (Kd = 16 microM). The chemical attachment of the 4-hydroxycinnamoyl group to the 6'-carbon of desglucouzarin significantly inhibits binding. This agrees with predictions that a 5'-methyl group on cardenolides fits the receptor site optimally for the porcine kidney enzyme. The 4-hydroxycinnamic ester was also found to be fluorescent.
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PMID:Inhibition of Na+,K+-ATPase by the cardenolide 6'-O-(E-4-hydroxycinnamoyl) desglucouzarin. 979 Sep 42

Gap junctions appear to be essential components of metazoan animals providing a means of direct means of communication between neighboring cells. They are sieve-like structures which allow cell-cell movement of cytosolic solutes below 1000 MW. The major role of gap junctions would appear to be homeostatic giving rise to groups of cells which act as functional units. Ductin is the major core component of gap junctions and recent structural data shows it to be a four alpha-helical bundle which fits particularly well into a low resolution model of the gap junction channel. Ductin is also the main membrane component of the vacuolar H+-ATPase that is found in all eukaryotes and it seems likely that the gap junction channel first evolved as a housing for the rotating spindle of these proton pumps. Because ductin protrudes little from the membrane, other proteins are required to bring cell surfaces close enough together to form gap junctions. Such proteins may include connexins, a large family of proteins found in vertebrates.
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PMID:Structure of the ductin channel. 1035 72

Na+,K(+)-ATPase activity was studied in neurones and neuroglia under conditions of convulsions caused by picrotoxin administration. Picrotoxin is a stimulant which causes convulsions by suppression of presynaptic inhibition. Na+,K(+)-ATPase activity in neuroglia was increased in convulsion, bat was not altered in neurones. It is possible, that glial Na+, K(+)-ATPase activity plays the role of neuronal ion's flows regulator and neuronal protector from convulsions. In recovery period ATPase activities in neuronal and neuroglial cells run up to control.
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PMID:[ATPase activity in neurons and neuroglia during convulsions induced by picrotoxin]. 1037 4


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