Electrophoretic analysis of multiple forms of myosin in fast-twitch and slow-twitch muscles of the chick

Abstract

A method is described for the electrophoretic analysis of intact myosin in polyacrylamide gel in a buffer system containing 0.02 M-pyrophosphate and 10% (vol/vol) glycerol, pH 8.8. In this system chicken skeletal-muscle myosins reveal 5 distinct electrophoretic components, 3 components from the fast-twitch posterior latissimus dorsi muscle and 2 slower-migrating components from the slow-twitch anterior latissimus dorsi muscle. The Ca2+-activated ATPase activity of myosin components was measured by densitometric scanning of the gel for the Ca3(PO4)2 precipitate formed during the ATPase reaction and subsequently for stained protein. Each component from the same muscle appears to have identical ATPase activity, but components from the fast-twitch muscle had an activity 2.2 times higher than those from the slow-twitch muscle. On re-electrophoresis in the same buffer system, individual fractions of fast-twitch myosin did not reproduce the 3-band pattern of the original myosin, but migrated at rates consistent with their original mobility. Analysis of the mobility of the 3 fast-twitch myosin components in gels of different concentrations suggests that they are not stable oligomers of each other. These components of fast-twitch myosin and slow-twitch myosin are isoenzymes of myosin.