Comparison of ATP binding in the active sites of (Na+ + K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase with low affinity to calcium from cardiac sarcolemma.
The chemical composition of the active sites of cardiac sarcolemmal (Na+ + K(+)-ATPase, Mg(2+)-ATPase and Ca(2+)-ATPase has not been determined definitely. The present study deals with investigation of the role of OH group in position two on the ribose moiety of the ATP molecule in its interaction with the specific ATP binding sites on the above ATPases. Experiments with application of ATP and deoxyATP (the OH group in position 2 on the ribose absent revealed that neither Ca(2+)-ATPase nor Mg(2+)-ATPase is able to distinguish between ATP and deoxyATP as substrates). This indicates that the OH group investigated may play a negligible role only in ATP binding and splitting by the latter ATPases. On the contrary, kinetic studies of Na+ + K(+)-ATPase activation by deoxyATP revealed that the latter compound is a considerably less suitable substrate for the enzyme than ATP. Consequently the OH group in position 2 on the ribose moiety proved to be important both for ATP binding in the active site and for proper substrate turnover by (Na+ + K(+)-ATPase interaction of the ATP binding site of heart sarcolemmal ATPases. Results of the experiments showed that Ca(2+)-ATPase and Mg(2+)-ATPase cannot distinguish between ATP and deoxyATP as substrates. Kinetic studies of (Na+ + K+)-ATPase activation by deoxyATP revealed that the latter is a considerably less good substrate for the enzyme than ATP. It means that the OH group in position two on the ribose moiety proved to be important for both binding of ATP in the active site and for proper substrate turnover by (Na+ + K+)-ATPase.
Monosíková
R
R
Institute for Heart Research, Slovak Academy of Sciences, Bratislava.
Breier
A
A
Ziegelhöffer
A
A
Sima
F
F
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D012508:Sarcolemma / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism
D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases; D000254:Sodium-Potassium-Exchanging ATPase; D002118:Calcium