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Query: EC:3.6.3.14 (ATP synthase)
 7,042 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The coupling-factor ATPases from photosynthetically grown Rhodopseudomonas palustris and Rhodopseudomonas sphaeroides were purified by the same procedure to homogeneity. Gel chromatography on Sephacryl S-300 Superfine shortened the process of purification and improved its yield. Solubilization of the ATPase from both bacteria was found to be dependent on a specific sonication treatment of the cell suspensions, indicating a very weakly bound F1-ATPase in R. palustris. Depleted chromatophores could be restored in photophosphorylation and membrane-bound ATPase activities by adding the solubilized ATPase protein. The purified enzymes did not show a markedly trypsin-stimulated or dithiothreitol-stimulated activity. Isoelectric focusing and chromatofocusing revealed isoelectric points of 5.0 for both F1-ATPases. The molecular weights were determined by gel chromatography plus high-performance liquid chromatography. Hence, we calculated a molecular weight of 350000 for both F1-ATPases. Sodium dodecylsulfate/polyacrylamide gel electrophoresis revealed five subunits for both enzymes. Kinetic parameters, regarding substrate specificity, the effect of divalent cations, Km and Ki values for the membrane-bound and solubilized ATPases were determined.
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PMID:Purification and properties of the coupling-factor ATPases F1 from Rhodopseudomonas palustris and Rhodopseudomonas sphaeroides. 621 69

Ca-ATPase is thought to function as a calcium extrusion pump that may regulate cytosolic calcium concentration. Because the parathyroid gland is among the few tissues that are directly regulated by extracellular calcium and because cytosolic calcium may be a mediator of the effects of extracellular calcium on parathyroid hormone secretion, we have investigated the presence of this enzyme in homogenates of parathyroid cells. High performance liquid chromatography (HPLC) was used to quantify the formation of ADP from ATP following incubation of ATP with cellular homogenate in a buffer containing ethylenedioxy- (diethylenedinitrilo) tetra acetic acid (EGTA), ouabain, and calcium. Enzyme activity was calcium-dependent, with Ca-ATPase showing two Km (Ca) values, 31 and 853 nM. High affinity Ca-ATPase activity was reduced by the calmodulin inhibitor, trifluoperazine (TFP), with half-maximal inhibition occurring at 7 X 10(-5) M. Monovalent cations stimulated high affinity Ca-ATPase activity (K+ greater than Na+ greater than Rb+ greater than Li+) in the presence of calcium. Magnesium (0.8 mM) also stimulated cleavage of ATP. Sodium increased Ca-dependent ATPase activity by 82% but had no significant effect on Mg-stimulated activity. Furthermore, azide, an inhibitor of mitochondrial ATPase(s), had a significantly greater inhibitory effect on Mg-dependent than on Ca-dependent activity. In summary, a high affinity Ca-ATPase is present in bovine parathyroid cells which has a Km in the range of the cytosolic calcium concentration that is found in other cells. Ca-ATPase(s) may be of importance in regulating the cytosolic calcium concentration and, therefore, hormonal secretion in this cell type.
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PMID:Ca-ATPase activity in bovine parathyroid cells. 622 2

ATP induced swelling of isolated yeast mitochondria suspended in an isoosmotic solution of potassium gluconate. Valinomycin stimulated the swelling rate, indicating that K+ influx in the presence of ATP is rate-controlling. This swelling was inhibited by ADP, phosphate (probably acting on the external face of the inner membrane), and Mg2+, which forms a complex with ATP. ATP-induced swelling did not require working F0-F1-ATPase since it was not inhibited by oligomycin and uncoupler. CTP and GTP also induced a swelling. ATP also induced mitochondrial swelling in potassium glutamate, chloride, and acetate but not in phosphate solutions. Sodium, but not ammonium, can replace potassium ion. It is probable that the ATP-channel opening also necessitates an electrogenic cation influx. Respiration also induced swelling of mitochondria suspended in isoosmotic potassium gluconate solution. ATP- or respiration-induced swelling were inhibited equally by N,N'-dicyclohexylcarbodiimide, propranolol, and Zn2+ but not by quinine; all these drugs inhibit the H+/K+ exchange. It was concluded that this unspecific channel is not open under conditions used to measure oxidative phosphorylation. Its physiological role remains unknown.
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PMID:ATP-induced unspecific channel in yeast mitochondria. 752 86

For many bacteria Na+ bioenergetics is important as a link between exergonic and endergonic reactions in the membrane. This article focusses on two primary Na+ pumps in bacteria, the Na(+)-translocating oxaloacetate decarboxylase of Klebsiella pneumoniae and the Na(+)-translocating F1Fo ATPase of Propionigenium modestum. Oxaloacetate decarboxylase is an essential enzyme of the citrate fermentation pathway and has the additional function to conserve the free energy of decarboxylation by conversion into a Na+ gradient. Oxaloacetate decarboxylase is composed of three different subunits and the related methylmalonyl-CoA decarboxylase consists of five different subunits. The genes encoding these enzymes have been cloned and sequenced. Remarkable are large areas of complete sequence identity in the integral membrane-bound beta-subunits including two conserved aspartates that may be important for Na+ translocation. The coupling ratio of the decarboxylase Na+ pumps depended on delta muNa+ and decreased from two to zero Na+ uptake per decarboxylation event as delta mu Na+ increased from zero to the steady state level. In P. modestum, delta mu Na+ is generated in the course of succinate fermentation to propionate and CO2. This delta mu Na+ is used by a unique Na(+)-translocating F1Fo ATPase for ATP synthesis. The enzyme is related to H(+)-translocating F1Fo ATPases. The Fo part is entirely responsible for the coupling of ion specificity. A hybrid ATPase formed by in vivo complementation of an Escherichia coli deletion mutant was completely functional as a Na(+)-ATP synthase conferring the E. coli strain the ability of Na(+)-dependent growth on succinate. The hybrid consisted of subunits a, c, b, delta and part of alpha from P. modestum and of the remaining subunits from E. coli. Studies on Na+ translocation through the Fo part of the P. modestum ATPase revealed typical transporter-like properties. Sodium ions specifically protected the ATPase from the modification of glutamate-65 in subunit c by dicyclohexylcarbodiimide in a pH-dependent manner indicating that the Na+ binding site is at this highly conserved acidic amino acid residue of subunit c within the middle of the membrane.
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PMID:Bacterial sodium ion-coupled energetics. 783 94

Previous attempts to isolate a stable F0F1-ATPase complex (H(+)-translocating ATPase) from Vibrio parahaemolyticus have been unsuccessful. Using new non-ionic detergents (alkyl thiomaltosides), a stable F0F1 complex with a high specific activity (15-25 units/mg protein) was purified and characterized. The purified F0F1-ATPase consists of eight subunits (alpha, beta, gamma, delta, epsilon, a, b and c). The new detergents, in combination with sucrose (or glycerol), lipid, dithiothreitol and phenylmethylsulfonyl fluoride, effectively stabilized the F0F1 complex. The ATPase activity of the F0F1 complex was greatly increased by anions, such as SO4(2-) and SO3(2-). Sodium ion increased the activity by about 2-fold. Dicyclohexylcarbodiimide, Zn2+, 4-acetamido-4'-isothiocyanostilben-2,2'disulfonate and tetrachlorosalicylanilide inhibited F0F1-ATPase activity. Ethanol, which stimulated F1-ATPase activity, inhibited F0F1-ATPase activity. Methanol, Na3VO4 and bafilomycin A1 did not have any significant effect on F0F1-ATPase activity, although methanol, like ethanol, stimulated F1-ATPase activity.
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PMID:F0F1-ATPase of Vibrio parahaemolyticus: purification using new detergents and characterization. 794 6

In the crystal structure of the membrane-embedded rotor ring of the sodium ion-translocating adenosine 5'-triphosphate (ATP) synthase of Ilyobacter tartaricus at 2.4 angstrom resolution, 11 c subunits are assembled into an hourglass-shaped cylinder with 11-fold symmetry. Sodium ions are bound in a locked conformation close to the outer surface of the cylinder near the middle of the membrane. The structure supports an ion-translocation mechanism in the intact ATP synthase in which the binding site converts from the locked conformation into one that opens toward subunit a as the rotor ring moves through the subunit a/c interface.
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PMID:Structure of the rotor ring of F-Type Na+-ATPase from Ilyobacter tartaricus. 1586 Jun 15