title: Soluble and particulate forms of muscle alkaline proteinase show differential sensitivity to endogenous inhibitor(s).
abstract: Membrane-free washed myofibrils derived from rat skeletal muscle homogenates contained a chymostatin-sensitive protease(s) which acted on associated myofibrillar proteins, at an optimum pH of 8.5, much less rapidly at low ionic strength (insoluble myofilaments) than at high salt concentrations (solubilized proteins). When the myofibrillar fraction was added to the particle-free cytosol prepared from the muscle extracts, proteins of the cytosol were also degraded, but the activity in this case was much more pronounced at low ionic strength. This was because inhibitor(s) of the proteinase present in the cytosol fraction were only effective at high ionic strength when all the myofibrillar (and associated) proteins were in solution. The protease was separated from the bulk of the myofibrillar proteins by gel chromatography at high ionic strength. On dialysis against a low-salt buffer, part of the enzyme was precipitated. The putative cytosolic inhibitor(s) were again only effective on the soluble enzyme at high ionic strength.
mesh_terms: D000818:Animals; D002364:Caseins / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004229:Dithiothreitol / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000032:analysis* / Q000378:metabolism; D066298:In Vitro Techniques; D009132:Muscles / Q000201:enzymology*; D009210:Myofibrils / Q000201:enzymology; D011480:Protease Inhibitors / Q000494:pharmacology*; D051381:Rats; D012697:Serine Endopeptidases
chemical_list: D002364:Caseins; D011480:Protease Inhibitors; C026018:azocasein; D004533:Egtazic Acid; D010450:Endopeptidases; D012697:Serine Endopeptidases; D004229:Dithiothreitol