Purification of messenger ribonucleic acids for fast and slow myosin heavy chains by indirect immunoprecipitation of polysomes from embryonic chick skeletal muscle.
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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通过对来自胚胎鸡骨骼肌的多核糖体进行间接免疫沉淀，纯化快速和慢速肌球蛋白重链的信使核糖核酸。
DOI：
10.1021/bi00550a035
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发表时间：
1980
期刊：
影响因子：
2.9
通讯作者：
M. Rabinowitz
中科院分区：
文献类型：
--
作者：
P. K. Umeda;R. Zak;M. Rabinowitz
采用间接免疫沉淀法从14日龄鸡胚腿肌中分离快肌球蛋白重链和慢肌球蛋白重链mRNA。制备的抗体对肌球蛋白重链纯化NaDod-SO 4-聚丙烯酰胺凝胶电泳，并显示出特异性的快速和慢速肌球蛋白重链。RNA组分指导在无细胞翻译系统中从小麦胚芽合成肌球蛋白重链。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，因为它们与肌球蛋白重链的抗体发生免疫沉淀。用快肌球蛋白重链和慢肌球蛋白重链的抗体对翻译产物进行免疫沉淀，结果表明，相对于肌球蛋白HC型，RNA制剂中快肌球蛋白重链mRNA约富集94%，慢肌球蛋白重链mRNA约富集84%。具有轻微不同的迁移率的肽NaDodSO 4-聚丙烯酰胺凝胶免疫沉淀的抗体快速和缓慢的肌球蛋白重链。
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.