Lazarides E, Lindberg U
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4742-6. doi: 10.1073/pnas.71.12.4742.
Various tissues and cells in culture contain a specific inhibitor of DNase I (EC 3.1.4.5). In this paper evidence is presented that this inhibitor is actin, one of the major structural proteins of muscle and nonmuscle cells. (a) The inhibitor is a major cellular component constituting 5-10% of the soluble protein. (b) It migrates with actin on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, having a characteristic molecular weight of 42,000. (c) It has an amino-acid composition closely similar to that of actin. (d) The peptide maps of the two proteins are nearly identical. (e) Skeletal muscle actin inhibits the enzymatic activity of DNase I. (f) DNase I-agarose affinity chromatography quantitatively retains purified skeletal muscle actin, and actin, specifically, from high-speed supernatants of whole cell extracts. (g) An antibody to purified inhibitor protein from calf thymus, used in indirect immunofluorescence on cells grown in culture, stains a two-dimensional network of fibers similar to that seen with an actin-specific antibody.The observation that actin can be isolated by DNase-agarose affinity chromatography provides a useful tool for the biochemical study of actin under different physiological conditions.
培养中的各种组织和细胞都含有一种脱氧核糖核酸酶I(EC 3.1.4.5)的特异性抑制剂。本文提供的证据表明,这种抑制剂是肌动蛋白,它是肌肉细胞和非肌肉细胞的主要结构蛋白之一。(a)该抑制剂是一种主要的细胞成分,占可溶性蛋白的5% - 10%。(b)它在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳中与肌动蛋白一起迁移,具有42,000的特征分子量。(c)它的氨基酸组成与肌动蛋白非常相似。(d)这两种蛋白质的肽图几乎相同。(e)骨骼肌肌动蛋白可抑制脱氧核糖核酸酶I的酶活性。(f)脱氧核糖核酸酶I - 琼脂糖亲和层析可定量保留纯化的骨骼肌肌动蛋白,特别是来自全细胞提取物高速上清液中的肌动蛋白。(g)用小牛胸腺纯化的抑制剂蛋白抗体对培养的细胞进行间接免疫荧光染色,可染出类似于用肌动蛋白特异性抗体观察到的二维纤维网络。通过脱氧核糖核酸酶 - 琼脂糖亲和层析可以分离肌动蛋白这一观察结果,为在不同生理条件下对肌动蛋白进行生化研究提供了一个有用的工具。