Myosin isozymes in avian skeletal muscles. II. Fractionation of myosin isozymes from adult and embryonic chicken pectoralis muscle by immuno-affinity chromatography

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Abstract
Chicken pectoralis consists primarily of large white fibres, which react exclusively with antibodies prepared against adult fast myosin. There is, however, a small region of uniformly red fibres which responds to antibodies against adult slow myosin as well as adult fast myosin. The myosin extracted from this red region is also heterogeneous as shown by the presence of both slow and fast light chains. By means of immunoadsorbents, it has been possible to separate the ‘red myosin’ into a ‘fast’ component and a ‘slow’ component. These two fractions have been characterized with respect to their light and heavy chain content by one-dimensional and two-dimensional gel electrophoresis. The myosin heavy chain was reduced to the smaller fragments required for electrophoresis by proteolytic degradation. We conclude from the electrophoretic patterns that the ‘fast’ and ‘slow’ myosin components from the pectoralis red region closely resemble the myosin from the white region of the pectoralis and the myosin from the slow anterior latissimus dorsi (ALD) muscle. The demonstration of a ‘slow myosin’ in adult pectoralis muscle raises the possibility that the crossreactivity of embryonic pectoralis myosin with anti-slow (ALD) myosin antibodies might be due to the presence of such slow components in embryonic chicken muscle. Direct isolation of a slow component from embryonic pectoralis was achieved by immunoadsorption, as described for adult mixed muscle myosin. Analysis of the subunit composition by gel electrophoresis shows an enrichment in adult-type slow light chains, but the heavy chain pattern is quite distinct from that of adult slow heavy chain. These studies suggest that several myosin isozymes exist in embryonic chicken pectoralis, but that none is identical to those myosins found in the different fibres of the adult pectoralis muscle.