[ATPase activity of the novocaine segregation zones isolated from the erythrocytes of the common frog].
Novocaine segregation zones in frog's erythrocytes, isolated by differential centrifugation, were shown to be ATPase active. The enzyme displays half of its maximum activity at 0.18 Mm ATP concentration to be inhibited by high concentrations of ATP. ATPase is activated by both Mg2+ and Ca2+ (in a lesser degree), with the maximum activity being at pH 7.5. A 5 minutes heating without the substrate results in decreasing the enzyme activity at 30 degrees, and in the total inhibition at 50 degrees C. Along with ATP, the enzyme can hydrolyse GTP and, in a lesser degree, ADP and sodium pyrophosphate. The ATPase activity is not effected with oligomycin (0.5-1.5 mkg/ml) or ouabaine (0.1 mM). Oligomycin in concentration 5 micrograms/ml induced non-specific inhibition of ATPase. Uncouplers, like 2,4-dinitrophenol and carbonyl cyanid p-trifluorometoxyphenylhydrazone, stimulate the enzyme activity. The lack in the ATP-ase sensitivity to oligomycin (specific inhibitor of mitochondrial F1-ATPase) and ouabaine (specific inhibitor of Na+, K+-ATPase) may suggest that the ATPase activity of novocaine segregation zones in frog's erythrocytes is not associated with a random contamination with mitochondria or cytoplasmic membranes. The ATPase under study has much in common with the lysosomal +H-ATPase. The results obtained support a hypothesis that +H-ATPase may function as a course of protones for maintaining acidic medium in segregation zones and promote accumulation of weak bases by means of their protonation.
Bulychev
A G
AG
Veselkina
M N
MN
Braun
A D
AD
D000251:Adenosine Triphosphatases / Q000097:blood*; D000818:Animals; D002458:Cell Fractionation; D002469:Cell Separation; D004789:Enzyme Activation / Q000187:drug effects; D004912:Erythrocytes / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D011343:Procaine / Q000097:blood*; D011522:Protons; D011896:Rana temporaria; D013379:Substrate Specificity / Q000187:drug effects
D011522:Protons; D011343:Procaine; D000251:Adenosine Triphosphatases
ATFaznaia aktivnost' zon segregatsii novokaina, izolirovannykh iz éritrotsitov travianoĭ liagushki.