Umeda P K, Zak R, Rabinowitz M
Biochemistry. 1980 Apr 29;19(9):1955-65. doi: 10.1021/bi00550a035.
Fast and slow myosin heavy chain mRNAs 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 NaDod-SO4-polyacrylamide gel electrophoresis and were shown to be 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 also 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 approximately 94% enriched for fast myosin heavy chain mRNA and approximately 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.
通过对14日龄鸡胚腿部肌肉多核糖体进行间接免疫沉淀,分离出快肌球蛋白重链和慢肌球蛋白重链mRNA。抗体是针对经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳纯化的肌球蛋白重链制备的,并且已证明对快肌球蛋白重链和慢肌球蛋白重链具有特异性。这些RNA组分在来自小麦胚芽的无细胞翻译系统中指导肌球蛋白重链的合成。还以较低浓度合成了几种较小的肽。这些可能是肌球蛋白重链的部分产物,因为它们能用抗肌球蛋白重链抗体进行免疫沉淀。用抗快肌球蛋白重链和慢肌球蛋白重链抗体对翻译产物进行免疫沉淀,结果表明,就肌球蛋白HC类型而言,RNA制剂中快肌球蛋白重链mRNA富集了约94%,慢肌球蛋白重链mRNA富集了约84%。在十二烷基硫酸钠-聚丙烯酰胺凝胶上迁移率略有不同的肽,能用抗快肌球蛋白重链和慢肌球蛋白重链抗体进行免疫沉淀。