Inhibition of actin-myosin subfragment 1 ATPase activity by troponin I and IC: relationship to the thin filament states of muscle.
Troponin I (TnI) is the component of the troponin complex that inhibits actomyosin ATPase activity, and Ca(2+) binding to the troponin C (TnC) component reverses the inhibition. Effects of the binding of TnI and the TnI-TnC (TnIC) complex to actin-tropomyosin (actinTm) on ATPase and on the binding kinetics of myosin subfragment 1 (S1) were studied to clarify the mechanism of the inhibition. TnI and TnIC in the absence of Ca(2+) bind to actinTm and inhibit ATPase to similar levels with a stoichiometry of one TnI or one TnIC per one Tm and seven actin subunits. TnI also binds to actinTmTn in the presence of Ca(2+) with a stoichiometry and inhibition constant similar to those for the binding to actinTm of TnI and Tn in the absence of Ca(2+). Thus, in the presence of Ca(2+), the intrinsic TnI which is released from its binding site on actinTm does not interfere with the binding of an extra molecule of TnI to actinTmTn. The rate of S1 binding to actinTmTnI and to actinTmTnTnI in the presence of Ca(2+) was inhibited to the same extent as upon removal of Ca(2+) from actinTmTn. These studies show that TnI inhibits ATPase by the same mechanism as Tn in the absence of Ca(2+), by shifting the thin filament equilibria from the open state to the closed and blocked states.
Department of Biosciences, University of Kent, Canterbury, UK. M.A.Geeves@ukc.ac.uk
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Geeves
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M
Chai
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Lehrer
D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000378:metabolism; D000205:Actomyosin / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001667:Binding, Competitive; D002118:Calcium / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D008956:Models, Chemical; D018482:Muscle, Skeletal / Q000201:enzymology*; D015879:Myosin Subfragments / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009218:Myosins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D011817:Rabbits; D013329:Structure-Activity Relationship; D019209:Troponin C / Q000737:chemistry* / Q000378:metabolism; D019210:Troponin I / Q000737:chemistry* / Q000378:metabolism
D004791:Enzyme Inhibitors; D015879:Myosin Subfragments; D019209:Troponin C; D019210:Troponin I; D000205:Actomyosin; D009218:Myosins; D002118:Calcium