Characterization of the membrane bound Mg2+-ATPase of rat skeletal muscle.
A procedure was developed to isolate a membrane fraction of rat skeletal muscle which contains a highly active Mg2+-ATPase (5-25 mumol Pi/mg min). The rate of ATP hydrolysis by the Mg2+-ATPase was nonlinear but decayed exponentially (first-order rate constant greater than or equal to 0.2 s-1 at 37 degrees C). The rapid decline in the ATPase activity depended on the presence of ATP or its nonhydrolyzable analog 5'-adenylyl imidodiphosphate (AdoPP[NH]P). Once inactivated, removal of ATP from the medium did not immediately restore the original activity. ATP- or AdoPP[NH]P-dependent inactivation could be blocked by concanavalin A, wheat germ agglutinin or rabbit antiserum against the membrane. Additions of these proteins after ATP addition prevented further inactivation but did not restore the original activity. Low concentrations of ionic and nonionic detergents increased the rate of ATP-dependent inactivation. Higher concentrations of detergents, which solubilize the membrane completely, inactivated the Mg2+-ATPase. Cross-linking the membrane components with glutaraldehyde prevented ATP-dependent inactivation and decreased the sensitivity of the Mg2+-ATPase to detergents. It is proposed that the regulation of the Mg2+-ATPase by ATP requires the mobility of proteins within the membrane. Cross-linking the membrane proteins with lectins, antiserum or glutaraldehyde prevents inactivation; increasing the mobility with detergents accelerates ATP-dependent inactivation.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D017301:Ca(2+) Mg(2+)-ATPase; D003208:Concanavalin A / Q000494:pharmacology; D003902:Detergents; D007425:Intracellular Membranes / Q000201:enzymology; D037102:Lectins / Q000494:pharmacology; D008297:Male; D008854:Microscopy, Electron; D009132:Muscles / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D014909:Wheat Germ Agglutinins
D003902:Detergents; D037102:Lectins; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase