title: Proteolytic separation of the actin-activatable ATPase site from the phosphorylation site on the heavy chain of Acanthamoeba myosin IA.
abstract: Previous work (Maruta, H., Gadasi, H., Collins, J. H., and Korn, E. D. (1978) J. Biol. Chem. 253, 6292-6300) had shown that phosphorylation of the heavy chain of Acanthamoeba myosin IA is required for actin activation of its Mg2+-ATPase activity and that, like the phosphorylation site, the catalytic site and the actin binding site are also on the heavy chain. We now show that limited digestion of phosphorylated myosin IA by subtilisin allows separation of the catalytically active peptide fragment from the phosphorylated peptide without any significant loss of actin-activated Mg2+-ATPase activity. A proteolytic fragment with full actin-activated Mg2+-ATPase activity has also been isolated from subtilisin digests of nonphosphorylated myosin IA, which, before proteolysis, did not have actin-activated Mg2+-ATPase activity. The simplest interpretation of these data is that, in its nonphosphorylated state, the phosphorylation site of Acanthamoeba myosin IA inhibits the catalytic site and that this inhibition can be reversed either by phosphorylation of the site or by proteolytically separating it from the catalytic site. Alternatively, phosphorylation and proteolysis may, by unrelated mechanisms, induce similar conformational changes in the myosin heavy chain that lead to activation of its actomyosin ATPase activity.
mesh_terms: D000199:Actins / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000656:Amoeba / Q000201:enzymology*; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D004789:Enzyme Activation; D007700:Kinetics; D046911:Macromolecular Substances; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D010766:Phosphorylation; D013381:Subtilisins
chemical_list: D000199:Actins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D013381:Subtilisins; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins