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)

The distribution of I-labelled luteinizing hormone (LH) was studied by autoradiography of the murine pregnant ovary 10 min after intravenous administration. The grains in autoradiograms were localized around the luteal cells. The pregnant ovary showed the highest uptake of I-labelled LH 30 min. after the injection. No similar accumulation of I-labelled follicle-stimulating hormone, I-labelled albumin of free I was obtained. The ribosomal fraction of the corpus luteum contained a slightly higher level of radioactivity than the other subcellular fractions after injection of I-labelled LH. Sucrose density gradient centrifugation of the luteal particle preparation resulted in an enrichment of radioactivity in particles containing Na+,K+-activated ATPase, a marker enzyme of the plasma membrane. These findings support the concept of plasma membrane binding as the initial event in LH action on the target tissue.
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PMID:Attachment to the luteal plasma membranes: an early event in the action of luteinizing hormone. 414 22

The mechanism of action of diphtheria toxin in an Escherichia coli cell-free protein-synthesizing system was examined. When the washed ribosomes were incubated with toxin before addition of messenger ribonucleic acid (RNA), peptide syntheses of (14)C-phenylalanine directed by polyuridylic acid or phage R17 RNA were strongly inhibited by a small amount of toxin. Whereas, if the soluble fraction (105,000 x g supernatant fraction) was preincubated with toxin, no effect of toxin occurred either on the induced protein synthesis or on the activity of guanosine triphosphatase even in the presence of nicotinamide adenine dinucleotide. The binding of (3)H-formylmethionyl-transfer RNA to E. coli ribosomes directed by either R17 RNA or trinucleotide AUG was also decreased by toxin. These findings suggest that diphtheria toxin may prevent the binding of messenger RNA by successfully competing with the AUG for ribosomal binding sites. Sucrose-density gradient studies support this concept by showing the decrease in binding of (3)H-labeled R17 RNA to E. coli ribosomes exposed to toxin.
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PMID:Inhibitory effect of diphtheria toxin on amino acid incorporation in Escherichia coli cell-free system. 431 20

Sugar transport and glycolysis in Streptococcus sanguis NCTC 7865, Streptococcus mitis ATCC 903, Streptococcus salivarius NCTC 8606 and several strains of Streptococcus mutans were investigated by following the rate of acid production by washed bacteria at a constant pH of 7.0. The phosphoenolpyruvate-phosphotransferase system (PTS) was inhibited by low concentrations of chlorhexidine. When this PTS-inhibitory concentration of chlorhexidine was added to cells washed and re-suspended in KCl, glucose uptake and glycolysis continued at a greatly-reduced rate. Chlorhexidine abolished glucose and sucrose uptake and metabolism in bacteria washed and incubated in saline. The Na+-inhibition was reproduced in KCl-washed bacteria using the cyclic peptide ionophores, valinomycin and gramicidin, to dissipate K+ and H+ gradients across the cell membrane. Glucose metabolism by Strep. mutans B13 was more resistant to chlorhexidine than that of Strep. mutans NCTC 10449 or Strep. sanguis but was more sensitive to the ionophores. Valinomycin had a greater inhibitory effect on strain B13 than the other two. That ion gradients are important in the chlorhexidine-resistant glucose-uptake mechanism was confirmed using the classical uncoupling agents, carbonylcyanide-m-chlorophenylhydrazone, 2,4-dinitrophenol and KSCN. Glucose metabolism was inhibited in the presence of both the uncouplers and the PTS-inhibitory concentration of chlorhexidine and significant inhibition was also observed in the absence of the PTS inhibitor. Lactate or the ATPase inhibitor, dicyclohexyl carbodiimide (DCCD), had similar inhibitory effects on the non-PTS uptake system.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Evidence that glucose and sucrose uptake in oral streptococcal bacteria involves independent phosphotransferase and proton-motive force-mediated mechanisms. 609 4

Using a vacuolar preparation virtually free of contamination by other organelles, we isolated vacuolar membranes and demonstrated that they contain an ATPase. Sucrose density gradient profiles of vacuolar membranes show a single peak of ATPase activity at a density of 1.11 g/cm3. Comparison of this enzyme with the two well-studied proton-pumping ATPases of Neurospora plasma membranes and mitochondria shows that it is clearly distinct. The vacuolar membrane ATPase is insensitive to the inhibitors oligomycin, azide, and vanadate, but sensitive to N,N'-dicyclohexylcarbodiimide (Ki = 2 microM). It has a pH optimum of 7.5, requires a divalent cation (Mg2+ or Mn2+) for activity, and is remarkably unaffected (+/- 20%) by a number of monovalent cations, anions, and buffers. In its substrate affinity (Km for ATP = 0.2 mM), substrate preference (ATP greater than GTP, ITP greater than UTP greater than CTP), and loss of activity with repeated 1 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid washes, the vacuolar membrane ATPase resembles the F1F0 type of ATPase found in mitochondria and differs from the integral membrane type of ATPase in plasma membranes.
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PMID:Identification and properties of an ATPase in vacuolar membranes of Neurospora crassa. 621 2

Adenylate cyclase was solubilized from washed particulate fraction of rabbit cerebral cortex with the nonionic detergent Lubrol 12A9 and subjected to either gel filtration on Ultrogel AcA 34 or chromatography on DEAE Bio-Gel A. By both procedures the enzyme was resolved into two components, one insensitive to guanyl 5'-yl imidodiphosphate [Gpp(NH)p] and NaF but stimulated by Ca2+ and calmodulin, and another that was sensitive to Gpp(NH)p and NaF but relatively insensitive to Ca2+ and calmodulin. The data support the possibility that two independent forms of adenylate cyclase exist in cerebral cortex, one regulated by guanine nucleotide regulatory protein and another by Ca2+-calmodulin. Fractions containing the guanylnucleotide-sensitive activity were found to contain a factor that inhibited basal and Ca2+-stimulated adenylate cyclase in the Ca2+-sensitive fraction. The inhibitor was inactivated by heating at 60 degrees C and by incubation with trypsin. Inhibition was not time-dependent, and it was not due to destruction of cAMP by phosphodiesterase or of ATP by ATPase. Inhibitory action was not reversed by calmodulin and therefore it does not appear to be a calmodulin binding protein. Sucrose density gradient sedimentation indicated a sedimentation coefficient of 4S for the inhibitor; by this technique it co-sedimented with the adenylate cyclase sensitive to Gpp(NH)p and NaF.
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PMID:Properties of detergent-dispersed adenylate cyclase from cerebral cortex. Presence of an inhibitor protein. 626 48

N-Ethylmaleimide was employed as a surface label for sarcolemmal proteins after demonstrating that it does not penetrate to the intracellular space at concentrations below 1.10(-4) M. The sarcolemmal markers, ouabain-sensitive (Na+ +K+)-ATPase and Na+/Ca2+-exchange activities, were inhibited in N-ethylmaleimide perfused hearts. Intracellular activities such as creatine phosphokinase, glutamate-oxaloacetate transaminase and the internal phosphatase site of the Na+ pump (K+-p-nitrophosphatase) were not affected. Almost 20% of the (Ca2+ +Mg2+)-ATPase and Ca2+ pump were inhibited indicating the localization of a portion of this activity in the sarcolemma. Sarcolemma purified by a recent method (Morcos, N.C. and Drummond, G.I. (1980) Biochim. Biophys. Acta 598, 27-39) from N-ethylmaleimide-perfused hearts showed loss of approx. 85% of its (Ca2+ +Mg2+-ATPase and Ca2+ pump compared to control hearts. (Ca2+ +Mg2+)-ATPase and Ca2+ pump activities showed two classes of sensitivity to vanadate ion inhibition. The high vanadate affinity class (K1/2 for inhibition approx. 1.5 microM) may be localized in the sarcolemma and represented approx. 20% of the total inhibitable activity in agreement with estimates from N-ethylmaleimide studies. Sucrose density fractionation indicated that only a small portion of Mg2+-ATPase and Ca2+-ATPase may be associated with the sarcolemma. The major portion of these activities seems to be associated with high density particles.
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PMID:Localization of (Ca2+ + Mg2+)-ATPase, Ca2+ pump and other ATPase activities in cardiac sarcolemma. 628 90

The effects of sucrose feeding on parameters associated with (Na+,K+)-ATPase in brown adipose tissue were compared in rats treated with parenteral 6-hydroxydopamine and vehicle. Sucrose feeding significantly increased K+-p-nitrophenylphosphatase and ouabain binding in brown adipose tissue from rats treated with vehicle. By contrast, sucrose feeding had no effects on these measurements in rats treated with parenteral 6-hydroxydopamine. 6-hydroxydopamine did not significantly alter sucrose consumption and there were no significant effects on weight gain during the short experimental period.
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PMID:(Na+,K+)-ATPase stimulation by sucrose feeding: prevention by 6-hydroxydopamine. 631 40

Several lines of evidence show a close association between plasma membrane Na,K-ATPase and mitochondrial respiration. Extending the observation in human erythrocyte membrane (6), Na,K-ATPase activity has been shown to be elevated in kidney microsomal preparations from protein- and energy-malnourished rats (10). Kidney mitochondrial respiration was studied in these rats under various conditions of assay. Sucrose was used as a modifier of mitochondrial morphology and volume to study its effect on these mitochondria. Mitochondrial state 3 respiration was increased by 35% in protein-deficient rats (P less than 0.02). Vmax(ADP) of state 3 respiration was increased by about 47% in protein- as well as energy-restricted rats. Mitochondria from protein- and energy-deficient rats were more tightly coupled as compared to those from control group. Km apparent for (ADP) and (Pi) were elevated in protein- and energy-malnourished rats. The magnitude of increase was much more in energy-deficient rats. Morphological differences between the mitochondria from two dietary manipulations were reflected in differences in the responses of state 3 respiration, Km(ADP), state 4 respiration, and respiratory control ratios to changing sucrose concentrations. This increase in mitochondrial respiration parallels the increased Na,K-ATPase activity in these rats. Increased Km (ADP and Pi) for mitochondrial respiration are perhaps in response to increased availability of these metabolites in the cytosol. The sucrose effect, in addition, distinguishes the morphological differences in mitochondrial membrane due to protein or energy deficiencies. In conclusion, these results, to a great extent, support an association between the activity of Na,K-ATPase and mitochondrial respiration. The study of mechanism(s) which could contribute to the enhancement of mitochondrial respiration will be of general importance to the understanding of regulation of mitochondrial oxidative phosphorylation, and is of particular interest to us.
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PMID:Kidney mitochondrial respiration in protein- and energy-malnourished rats. 633 20

Extracellular matrix vesicles from rat alveolar bone were isolated by collagenase digestion and differential centrifugation. Further purification was performed by discontinuous sucrose density gradient centrifugation. Control tissues, kidney and liver, were processed according to the same procedures. Sucrose density gradient centrifugation of bone matrix vesicles revealed two peaks of enzymatic activity: "light" and "heavy" vesicle-enriched fractions. Electron micrographs revealed a higher degree of purification of the "light" rather than the "heavy" vesicle-enriched fraction. This coincided with the high levels of enzymatic activity detected in this fraction. Preparations obtained from kidney and liver had significantly lower levels of activity of alkaline phosphatase and ATPase as compared to the bone matrix vesicle fractions. There were also differences in the positions of enzyme activity peaks in the sucrose gradient fractions from the three tissues studied. Electron microscopic examination of kidney and liver fractions revealed structures larger than the purified bone matrix vesicles. In addition no electron-dense material was found within organelles from kidney and liver and they were studded with numerous ribosomes. Our observations indicate that the present method of isolation and purification yields fractions of matrix vesicles which are specific to bone and are significantly different from those obtained from kidney and liver.
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PMID:Purification and further characterization of isolated matrix vesicles from rat alveolar bone. 734 98

Potassium accumulation in rat heart after alpha-1-adrenoceptor stimulation has previously been reported from indirect measurements. Here we present data on intracellular potassium content measured directly in the heart. Isolated rat hearts perfused in a non-recirculating system were exposed to alpha-1-adrenoceptor stimulation (5 x 10(-5) mol/l phenylephrine in the presence of 10(-6) mol/l timolol). 14C-Sucrose was used to estimate the extracellular space. From heart homogenates intracellular potassium, magnesium and cellular water contents were determined and the ion concentrations calculated accordingly. The intracellular magnesium content remained unchanged during all experimental conditions. alpha-1-Adrenoceptor stimulation evoked an increase in potassium content by 9% (4, 14; 95% confidence interval (CI), P = 0.0006). Due to an observed increase in intracellular water by 17% (9, 26; 95% CI, P = 0.0006), the potassium concentration apparently decreased by 8% (0.3, 15; 95% CI, P = 0.04). During partial inhibition of the Na+/K(+)-ATPase by 10(-5) mol/l ouabain, there was an increase in potassium content by 5% (1, 9; 95% CI, P = 0.008). There was, however, no significant increase in intracellular water in this situation. Calculated intracellular potassium concentration showed accordingly a slight increase. The effects upon potassium and water both in the absence and presence of ouabain were eliminated by the alpha-1-adrenoceptor blocker prazosin (10(-6) mol/l). alpha-1-Adrenoceptor stimulation apparently increased cellular dry weight by 10% (2, 18; 95% CI, P = 0.02). Changes in translocation of potassium and water must be considered as part of the alpha-1-adrenergic heart effects.
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PMID:Increased intracellular potassium and water contents in rat heart after alpha-1-adrenoceptor stimulation. 787 Jul 1


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