Isolation and purification of actomyosin ATPase from mammalian brain.
A simple technique for the isolation and purification of mammalian brain actomyosin, based on extraction of whole brains in low ionic-strength buffer, is described. The final preparation of brain actomyosin is obtained in good yield, has relatively high K+-EDTA and Ca2+-ATPase activities, and is substantially free of other ATPases and tubulin. The preparation is useful for initial enzymatic studies and/or as an enrichment step toward purification of the individual protein components. The Mg2+-ATPase and K+-EDTA ATPase activities are strongly inhibited by the sulfhydryl blocking reagent, pHMB. Interaction between the actin and myosin components can be demonstrated. Brain actomyosin had a distinct electrophoretic profile and enzymatic activity when compared with smooth muscle actomyosin from the aorta.
D000199:Actins / Q000378:metabolism; D000205:Actomyosin / Q000302:isolation & purification; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001011:Aorta; D001921:Brain / Q000201:enzymology*; D004347:Drug Interactions; D004591:Electrophoresis, Polyacrylamide Gel; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D009130:Muscle, Smooth / Q000201:enzymology; D009218:Myosins / Q000378:metabolism; D011817:Rabbits
D000199:Actins; D006902:Hydroxymercuribenzoates; C030601:4-hydroxymercuribenzoate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins