Activation of skeletal S-1 ATPase activity by actin-tropomyosin-troponin. Effect of Ca++ on the fluorescence transient.
Regulation in striated muscles primarily involves the effect of changes in the free calcium concentration on the interaction of subfragment-1 (S-1) with the actin-tropomyosin-troponin complex (henceforth referred to as [acto]R). At low concentrations of free Ca++ the rate of ATP hydrolysis by (acto)R S-1 can be as much as 20-fold lower than that in the presence of high free Ca++, even though the binding of S-1 to (actin)R in the presence of ATP is virtually independent of the calcium concentration. This implies that the mechanism of regulation involves a kinetic transition between actin-bound states, rather than the result of changes in actin binding. In the current work, we have investigated the fluorescence transient that occurs with the binding and hydrolysis of ATP both at low and high free [Ca++]. The magnitude of this transition at low free [Ca++] is higher than at high free [Ca++]. At low free [Ca++], the rate of the fluorescence transient either stays constant or decreases slightly with increasing free actin concentrations, but at high free [Ca++] the rate increases slightly with increasing free actin concentration. The observed changes in rate are not great enough to be of regulatory importance. The results of the fluorescence transient experiments together with the binding studies performed at steady state also show that neither the binding of M.ATP or M.ADP.Pi to (actin)R is appreciably Ca++ sensitive. These data imply that an additional step (or steps) in the ATPase cycle, i.e., other than the burst transition, must be regulated by calcium.
Stein
L A
LA
Department of Medicine, State University of New York, Stony Brook.
Chalovich
J M
JM
D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D004789:Enzyme Activation; D066298:In Vitro Techniques; D007700:Kinetics; D009132:Muscles / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D014335:Tropomyosin / Q000378:metabolism; D014336:Troponin / Q000378:metabolism
D000199:Actins; D014335:Tropomyosin; D014336:Troponin; D000255:Adenosine Triphosphate; D009218:Myosins; D002118:Calcium