Preparation of Myosin Heavy Chain from Fish Muscle

Abstract

Myosin heavy chain (light chain-free myosin) was prepared from the white muscle of mackerel, striped pigfish, carp. jack mackerel and yellow tail by the modification of the method for rabbit. Myosin was treated with 2M LiCl-0.1M KCl-0.1M glycine buffer (pH 11.35) containing 1 mM ATP and 0.5% (v/v) 2-mercaptoethanol, for 50 min under reduced pressure. Crued myosin heavy chain was precipitated by adding 2.5M K citrate (pH 7.1), then purified by repeated dissolutions and precipitations.
Purities of the products were calculated to be 80-94% from the area of the band in the densitogram pattern for SDS polyacarylamide gel electrophoresis and their yields were 54-70% based on the myosin. The intrinsic viscosities of the heavy chains were 1.8dl/g for carp and 1.9dl/g for yellow tail at 10°C. E_??_ values at 280 nm were varied from 8 to 31 for the above fifhes at pH 7.5, and CD spectrum of the heavy chain from striped pigfish showed two extreme at 208 nm (θ=-32, 000) and 222 nm (θ=-36, 000).