Gene/Protein Disease Symptom Drug Enzyme Compound
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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)

When the bovine mitochondrial F1-ATPase is inactivated with dicyclohexyl[14C]carbodiimide and then gel-filtered, from 2 to 3 g atoms of 14C are incorporated/mol of enzyme. Prior inactivation of the enzyme by the modification of an essential tyrosine residue with 4-chloro-7-nitrobenzofurazan, a reaction that can be reversed by thiols, does not affect the irreversible inactivation of the ATPase by dicyclohexyl[14C]carbodiimide. During the large scale modification of the F1-ATPase by dicyclohexyl[14C]carbodiimide which led to 70% inactivation, 1.9 g atoms of 14C were incorporated/mol of enzyme. Isolation of the alpha, beta, and gamma subunits from this large scale inactivation revealed that the gram atoms of 14C bound per mol of each of the subunits was: alpha, 0.04; beta, 0.56; and gamma, 0.04. The majority of the radioactivity in a cyanogen bromide digest of the 14C-labeled beta subunit was isolated in a fragment that has the following amino acid sequence: Glu-Leu-Ile-Asn-Asn-Val-Ala-Lys-Ala-His-Gly-Gly-Tyr-Ser-Val-Phe-Ala-Gly-Val-Gly -Glu-Arg-Thr-Arg-Glu-Gly-Asn-Asp-Leu-Tyr-Glu*-His-Met; where Glu* represents the N gamma-glutamyl derivative of dicyclohexyl[14C]urea.
J Biol Chem 1981 Sep 10
PMID:Inactivation of the bovine mitochondrial F1-ATPase with dicyclohexyl[14C]carbodiimide leads to the modification of a specific glutamic acid residue in the beta subunit. 611 57

A previously unstudied acyl-coenzyme A thioesterase activity has been demonstrated in submitochondrial particles from Saccharomyces cerevisiae. The preferred substrate for the enzyme activity is oleoyl-coenzyme A. Tests with inhibitors of the thioesterase showed that, in addition to common thiol inhibitors, the oxidative phosphorylation inhibitors oligomycin and venturicidin also blocked thioesterase activity. Purification of the enzyme catalyzing this activity revealed that thioesterase copurified with mitochondrial ATPase. When thioesterase was isolated from oxidative phosphorylation mutants selected for resistance to these two inhibitors, thioesterase activity was also resistant. The results suggest that thioester hydrolysis may be catalyzed by components associated with the isolated ATPase complex. Further attempts to link this activity to in vivo function of ATPase were not successful.
Arch Biochem Biophys 1983 Sep
PMID:An acyl-thioesterase from yeast mitochondria. 613 97

The effects of tertiary amine local anesthetics (procaine, lidocaine, tetracaine and dibucaine) and chlorpromazine were investigated for three enzyme activities associated with rat brain synaptosomal membranes, i.e., (Na+ + K+)-ATPase (ouabain-sensitive), Mg2+-ATPase (ouabain-insensitive) and acetylcholinesterase. Approximately the same concentrations of each agent gave 50% inhibition of both ATPases, for example 7.9 and 10 mM tetracaine for Mg2+-ATPase and (Na+ + K+)-ATPase, respectively; these concentrations are 10-fold higher than required for inhibition of mitochondrial F1-ATPase. The relative inhibitory potency of the several agents was proportional to their octanol/water partition coefficients. Acetylcholinesterase was inhibited by all agents tested, but the ester anesthetics (procaine and tetracaine) were considerably more potent than the others after correction for partition coefficient differences. For tetracaine, 0.18 mM gave 50% inhibition and showed competitive inhibition on a Lineweaver-Burk plot, but for dibucaine a mixed type of inhibition was observed, and 0.63 mM was required for 50% inhibition. Tetracaine evidently binds at the active site, and dibucaine at the peripheral or modulator site, on this enzyme.
Biochim Biophys Acta 1984 Sep 07
PMID:Inhibition of synaptosomal enzymes by local anesthetics. 614 63

1. Antibodies prepared against the Rhodospirillum rubrum F1-ATPase (RrF1) and its purified, native-beta-subunit, exhibited cross-reactivity with the following soluble preparations of R. rubrum ATPase: RrF0 . F1, RrF1 and the beta-subunit. Anti-RrF1, but not anti-beta antibodies, also formed precipitin lines with soluble beta-less Rrf1, indicating that antigenic determinants of both the beta-subunit and the other four RrF1-subunits are expressed in the whole RrF1 molecule. Both antibodies agglutinated the R. rubrum chromatophores, suggesting that the beta-subunit is located on the external part of RrF1. 2. Both antibodies inhibited ATP synthesis and hydrolysis activities of R. rubrum chromatophores, as well as all the soluble ATPase reactions. Similar concentrations of each antibody were required for 50% inhibition of all these reactions, but anti-RrF1 was always somewhat more effective than anti-beta. These data indicate that the beta-subunit is involved in the catalytic site of the RrF1-enzyme. 3. The antibodies prepared against R. rubrum F1-ATPase and its beta-subunit could bind the soluble chloroplast F1-ATPase (CF1) and inhibited ATP-linked reactions carried out by chloroplasts and by soluble CF1. In these reactions, unlike in the R. rubrum ones, anti-beta was a more potent inhibitor than the anti-RrF1 antibody. The cross-reaction obtained between the antibodies raised against R. rubrum F1 and its beta-subunit and the chloroplast CF1 indicates the presence of similar antigenic determinants in the photosynthetic prokaryotic and eukaryotic F1-ATPases, which have been conserved during evolution.
Eur J Biochem 1981 Sep
PMID:Antibodies to the F1-ATPase of Rhodospirillum rubrum and its purified native beta-subunit: inhibition of ATP-linked activities in R. rubrum and in lettuce. 621 May 25

Purified F1-ATPase from Micrococcus lysodeikticus contains zinc in the amount of 1 mol/mol of enzyme. This zinc content correlates with standard values of ATPase activity (assayed with Ca2+-ATP as substrate) of the protein, i.e. 5--6 mumol substrate hydrolysed . min-1 . mg-1. Prolonged dialysis against EDTA results in a zinc-free protein which concomitantly loses its ATPase activity. Chelators such as Zincon, EDTA and L-cysteine inhibit the ATPase activity in concentration and/or time dependence related to their affinity for the metal ion involved. Reconstitution of the metallo (Zn2+) protein is demonstrated by the incorporation to the zinc-free protein of 65Zn2+ in amount near the 1 mol/mol of enzyme. This incorporation was concomitant with the regain of ATPase activity. The inhibition by EDTA and Zincon is reversed specifically by Zn2+ while the inhibition by EDTA is prevented by Zn2+ and Mn2+ and to, a minor extent, by Cd2+. Zn2+ and Ca2+ ions are involved and are probably mandatory in the ATPase activity of M. lysodeikticus F1 but their roles appear to be different and not exchangeable. Other divalent metal ions inhibit the Ca2+-ATPase activity of the Zn2+ protein by the following decreasing order; Hg2+, Fe2+, Co2+, Cd2+, Mn2+, Mg2+. M. lysodeikticus F1-ATPase is thus identified as a metallo (zinc) protein, which requires additional divalent metal ions for ATP hydrolysis.
Eur J Biochem 1981 Sep
PMID:Identification of a bacterial energy-transducing ATPase as a metallo (Zn2+) protein. Effect of chelating agents and divalent metal ions on ATPase activity. 621 May 27

N,N'-Dicyclohexylcarbodiimide (DCCD) covalently binds to the beta subunit of Escherichia coli F1-ATPase (BF1). The ATPase activity is fully inhibited when 1 mol of DCCD is bound/mol of BF1, in spite of the fact that BF1 contains several beta subunits [Satre, M., Lunardi, J., Pougeois, R., & Vignais, P.V. (1979) Biochemistry 18, 3134-3140]. Advantage was taken of the reactivity of DCCD with respect to BF1 to determine the exact stoichiometry of the beta subunits in BF1. Two methods were used. The first one was based on the fact that modification of the beta subunit by DCCD results in the disappearance of one negative charge, due to the binding of DCCD to a carboxyl group of the beta subunit. The nonmodified and the modified beta subunits were separated by electrofocusing, and the percentage of modified beta subunits was assessed as a function of the percentage of ATPase inactivation. The second method relied on direct comparison, after inactivation of BF1 by [14C]DCCD, of the specific radioactivities of the whole BF1 and the isolated beta subunits. Both methods indicate that each molecule of BF1 contains three beta subunits.
Biochemistry 1982 Sep 14
PMID:Chemical modification of F1-ATPase by dicyclohexylcarbodiimide: application to analysis of the stoichiometry of subunits in Escherichia coli F1. 621 38

The recently described methodology to extract the mitochondrial ATPase along with other mitochondrial proteins into organic solvents, and their subsequent incorporation into liposomes [Eur. J. Biochem. (1982) 121, 427-433] has been employed to estimate the number of ATPases that contain the natural ATPase inhibitor protein in its inhibitory site in intact mitochondria incubated in various metabolic states. It was found that in the presence of electrochemical gradients about 50% of the ATPases are without inhibitor protein in its inhibitory site (active ATPases). In the transition from state 4 to state 3 the percentage of active ATPases diminishes from about 50% to approximately 20%. This indicates that during steady-state phosphorylation only a limited number of ATPases are in the active catalytic state, and that not only during active hydrolysis does the inhibitor protein interact with its inhibiting site; rather the inhibitor seems to interact with an intermediate state of the enzyme that appears either during the synthetic or hydrolytic reactions. In addition it was found that ATP, with or without uncoupler, induces the interaction of the inhibitor protein in more than 80% the ATPases through an oligomycin-sensitive process. Thus, notwithstanding other factors the interaction may account for the low hydrolytic activity of mitochondria.
Eur J Biochem 1982 Sep 01
PMID:On the function of the natural ATPase inhibitor protein in intact mitochondria. 621 3

A method to optimize enzymatic assays by using pyruvate kinase and lactate dehydrogenase enzymes is presented and applied to mitochondrial ATPase as an example. Optimum amounts of auxiliary enzymes, to obtain either a 99% of the initial rate in a given time (t99) or a given lag period (L), are calculated from their apparent Michaelis constants (Kapp) in the medium used and their prices per enzymatic international unit.
Rev Esp Fisiol 1982 Sep
PMID:Enzymatic assays: optimization of systems by using pyruvate kinase and lactate dehydrogenase as auxiliary enzymes. 621 43

Antibody raised against the N,N'-dicyclohexylcarbodiimide (DCCD)-binding polypeptide of Escherichia coli bound to the cytoplasmic surface of the cell membrane. A weak reaction was seen with everted vesicles of the thermophile PS3. Rat-liver mitochondrial membranes did not react with the antibody. Reaction of the isolated DCCD-binding polypeptide with the antibody was prevented by oxidation of methionine residues or cleavage of the polypeptide with cyanogen bromide. Modification of the arginine residues of the DCCD-binding polypeptide did not affect interaction with the antibody. Purified F1-ATPase of E. coli bound to the isolated DCCD-binding polypeptide as shown by solid-phase radioimmune assay. Binding involved the alpha and/or beta subunits of F1 and the arginine residues of the polar central region of the DCCD-binding polypeptide. Our results are consistent with a looped arrangement of the DCCD-binding polypeptide in the membrane in which the carboxyl- and amino-terminal regions of the molecule are at the periplasmic surface and the polar central region, interacting with F1, is at the cytoplasmic surface of the cell membrane.
Biochim Biophys Acta 1983 Sep 07
PMID:Interaction of Escherichia coli F1-ATPase with dicyclohexylcarbodiimide-binding polypeptide. 622 13

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.
Metabolism 1983 Sep
PMID:Ca-ATPase activity in bovine parathyroid cells. 622 2


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