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Query: EC:3.6.3.1 (Mg2+-ATPase)
 1,484 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We have partially purified active delta and epsilon subunits of the E. coli membrane-bound Mg2+-ATPase (ECF1). Treating purified ECF1 with 50% pyridine precipitates the major subunits (alpha, beta, and gamma) of the enzyme, but the two minor subunits (delta and epsilon), which are present in relatively small amounts, remain in solution. The delta and epsilon subunits were then resolved from one another by anion exchange chromatography. The partially purified epsilon strongly inhibits the hydrolytic activity of ECF1. The epsilon fraction inhibits both the highly purified five-subunit ATPase and the enzyme deficient in the delta subunit. The latter result indicates that the delta subunit is not required for inhibition by epsilon. By contrast, two-subunit enzyme, consisting chiefly of the alpha and beta subunits, was insensitive to the ATPase inhibitor, suggesting that the gamma subunit may be required for inhibition by epsilon. The partially purified delta subunit restored the capacity of ATPase deficient in delta to recombine with ATPase-depleted membranes and to reconstitute ATP-dependent transhydrogenase. Previously we reported (Biochem, Biophys. Res. Commun. 62:764 [1975]) that a fraction containing both the delta and epsilon subunits of ECF1 restored the capacity of ATPase missing delta to recombine with depleted membranes and to function as a coupling factor in oxidative phosphorylation and for the energized transhydrogenase. These reconstitution experiments using isolated subunits provide rather substantial evidence that the delta subunit is essential for attaching the ATPase to the membrane and that the epsilon subunit has a regulatory function as an inhibitor of the ATPase activity of ECF1.
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PMID:Partial purification of active delta and epsilon subunits of the membrane ATPase from escherichia coli. 12 87

Effects in vitro of benthiocarb on rat brain Mg2+-ATPase were studied to elucidate the interaction of benthiocarb with Mg2+-ATPase. Based on IC50 values, it was evident that Mg2+-ATPase was sensitive to benthiocarb. Non-competitive inhibition with respect to activation by ATP was indicated by decreased maximal velocity (V) without change in Michaelis Menten constant (Km). It was also noted that pyridine-2-aldoxime (30 microM), pyridine-4-aldoxime (35 microM) and L-cysteine (90 microM) neutralized the inhibition of benthiocarb (8 microM).
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PMID:Kinetics of Mg2+-ATPase inhibition by benthiocarb and its recovery by oximes and L-cysteine in brain of albino rat (Wistar strain). 247 98

We found that isolated gastric vesicles contain a novel Mg2+-ATP-dependent phospholipid translocation (flippase) activity. Fluorescence analogue of phosphatidylcholine, 2-(12-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)dodecanoyl-1-hexadecanoyl-sn-glycero-3- phosphocholine, was ATP-dependently translocated from the outer (cytosolic) to inner (luminal) leaflet of the lipid membrane bilayer of hog gastric vesicles. The translocation was saturable and depended on time and the ATP concentration (Km = 3.1 microM). The basal Mg2+-ATPase activity of gastric vesicles in the absence of K+ showed high (Km = 1.6 microM) and low (Km = 80 microM) affinities for ATP, indicating that the present flippase activity is driven mostly by the high affinity Mg2+-ATPase activity. It required Mg2+ but not K+. Verapamil, which is an inhibitor of mouse mdr2 phosphatidylcholine flippase, did not inhibit the present flippase activity. Isolated sarcoplasmic reticulum vesicles that contain Ca2+-ATPase did not show any flippase activity. Fluorescence analogues of phosphatidylserine and phosphatidylethanolamine were similarly translocated by the gastric flippase. These phospholipid flippase activities were inhibited by 2-methyl-8-(phenylmethoxy)imidazo[1,2-a]pyridine-3-acetonitrile (SCH 28080) (IC50 = 0.14-0.25 microM), a specific K+-ATPase inhibitor of gastric H+,K+-ATPase rich in gastric vesicles. IC50 value for the SCH 28080-inhibitable Mg2+-ATPase activity was about 0.13 microM, indicating that the phospholipid translocation was driven mostly by the SCH 28080-sensitive Mg2+-ATPase activity. Possible physiological roles of flippases were discussed in relation with the gastric acid secretory and cytoprotective mechanisms.
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PMID:The phospholipid flippase activity of gastric vesicles. 909 84

Recently, a gastric Mg(2+)-ATP-dependent phospholipid flippase was found. Here, the effects of ionophores and monovalent cations on the gastric flippase were examined. We found that translocation of the fluorescent analogue of phosphatidylcholine was inhibited by valinomycin in the presence of K(+). The inhibition depended on both the concentrations of valinomycin and K(+). Valinomycin did not inhibit translocation in the absence of K(+). Protonophores, carbonylcyanide-m-chlorophenylhydrazone (CCCP) and carbonylcyanide-p-(trifluoromethoxy)phenylhydrazone (FCCP), accelerated translocation by 190-270%. These increases were completely abolished by 2-methyl-8-(phenylmethoxy)imidazo-[1, 2-a]pyridine-3-acetonitrile (SCH 28080), a gastric flippase inhibitor. Since these protonophores did not affect the Mg(2+)-dependent ATPase activity that is responsible for phospholipid translocation by the flippase, the coupling ratio of the amount of transported phospholipids/the amount of hydrolyzed ATP was variable and seemed to depend on the state of the membrane bilayer, for example fluidity. Inhibition by the valinomycin-K(+) complex was abolished in the presence of CCCP or FCCP, indicating the valinomycin-K(+)-CCCP(FCCF) ternary complex did not inhibit the flippase.
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PMID:Effects of ionophores on the phospholipid flippase activity of gastric vesicles. 1060 27