Effects of deuterium oxide on elementary steps in the ATPase reaction. Evidence for the similarity of key intermediates in contractile and transport ATPase.
The effects of D2O on the elementary steps in the contractile and transport ATPase [EC 3.6.1.3] reactions were studied, and the following results were obtained: 1. The rate of H-meromyosin ATPase in the steady state decreased in D2O to 60% of that in H2O. Deuterium oxide did not affect the size or rate of the initial burst of Pi liberation, i.e. the amount or rate of formation of the reactive myosin-phosphate-ADP complex, MADPP. Moreover, neither the rate of change in the fluorescence spectrum of H-meromyosin induced by ATP (the rate of formation of the second enzyme-ATP complex, M2ATP) nor the rate constant of decomposition of MADPP into M degrees + ADP + Pi was affected by D2O. However, the equilibrium constant of the step M2ATP in equilibrium MADPP decreased in D2O to about 1/2 the value in H2O. 2. In the case of the Na+-K+-dependent ATPase reactin, neither the rate constant of formation of the second enzyme-ATP complex, E2ATP, nor that of decomposition of a phosphorylated intermediate, EADP approximately P, was affected by D2O. However, the equilibrium constant of the step E2ATP in equilibrium EADP approximately P decreased in D2O to about 1/2.5-1/4 of the value in H2O. These results suggest a similarity between the modes of binding of phosphate in MADPP in the myosin ATPase reaction and in EADP approximatley P in the Na+-K+-dependent ATPase reaction.
D000199:Actins / Q000378:metabolism*; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D003903:Deuterium; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008433:Mathematics; D009119:Muscle Contraction; D009132:Muscles / Q000201:enzymology*; D015879:Myosin Subfragments / Q000378:metabolism*; D009928:Organ Specificity; D009994:Osmolar Concentration; D011189:Potassium Chloride / Q000494:pharmacology; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology
D000199:Actins; D015879:Myosin Subfragments; D012965:Sodium Chloride; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D003903:Deuterium; D000251:Adenosine Triphosphatases