Purification of messenger ribonucleic acids for fast and slow myosin heavy chains by indirect immunoprecipitation of polysomes from embryonic chick skeletal muscle

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Abstract
Fast and slow myosin heavy chain mRNA were isolated by indirect immunoprecipitation of polysomes from 14 day old embryonic chick leg muscle. The antibodies were prepared against myosin heavy chains purified by NaDodSO4[sodium dodecylsulfate]-polyacrylamide gel electrophoresis and were specific for fast and slow myosin heavy chains. The RNA fractions directed the synthesis of myosin heavy chains in a cell-free translation system from wheat germ. Several smaller peptides were synthesized in lower concentrations. These probably are partial products of myosin heavy chains, since they are immunoprecipitated with antibodies to myosin heavy chains. Immunoprecipitation of the translation products with the antibodies to fast and slow myosin heavy chains showed the RNA preparations to be .apprx. 94% enriched for fast myosin heavy chain mRNA and .apprx. 84% enriched for slow myosin heavy chain mRNA with respect to myosin HC type. Peptides having slightly different mobilities on NaDodSO4-polyacrylamide gels were immunoprecipitated by antibodies to fast and slow myosin heavy chains.