Hammer J A, Korn E D, Paterson B M
J Biol Chem. 1984 Sep 25;259(18):11157-9.
Addition of a myosin IB polyclonal antibody, which reacts with the heavy chains of both myosins IA and IB, to the [35S]methionine-labeled proteins synthesized in vitro using Acanthamoeba messenger RNA specifically immunoprecipitated radioactive polypeptides that migrated on sodium dodecyl sulfate-polyacrylamide gels exactly with the heavy chains of purified myosins IA and IB. A myosin II polyclonal antibody specifically immunoprecipitated a radioactive polypeptide which migrated exactly with the heavy chain of myosin II. These results provide strong evidence that Acanthamoeba myosins IA, IB, and II are separate gene products and that the purified proteins contain native, undegraded heavy chains. These results are especially important for myosins IA and IB, because they possess unusually small heavy chains for myosins.
向利用棘阿米巴信使核糖核酸体外合成的[35S]甲硫氨酸标记蛋白中添加一种肌球蛋白IB多克隆抗体(该抗体可与肌球蛋白IA和IB的重链发生反应),能特异性免疫沉淀放射性多肽,这些多肽在十二烷基硫酸钠-聚丙烯酰胺凝胶上的迁移情况与纯化的肌球蛋白IA和IB的重链完全一致。一种肌球蛋白II多克隆抗体特异性免疫沉淀了一种放射性多肽,其迁移情况与肌球蛋白II的重链完全一致。这些结果提供了有力证据,表明棘阿米巴肌球蛋白IA、IB和II是不同的基因产物,且纯化的蛋白包含天然的、未降解的重链。这些结果对肌球蛋白IA和IB尤为重要,因为它们的重链对于肌球蛋白来说异常小。