来自鸡胗的细丝蛋白（一种肌动蛋白结合蛋白）的纯化及特性

Purification and properties of filamin, and actin binding protein from chicken gizzard.

作者信息

Shizuta Y, Shizuta H, Gallo M, Davies P, Pastan I

出版信息

J Biol Chem. 1976 Nov 10;251(21):6562-7.

Abstract

Filamin, a protein recently identified in chicken gizzard (Wang, K., Ash, F., and Singer, S. J. (1975) Proc. Natl. Acad. Sci. U. S. A. 72, 4483-4486), has been purified free of other components and its molecular properties have been examined. Filamin has a sedimentation constant (S020,w) of 8.86 S and a partial specific volume of 0.734 ml/g. Sedimentation equilibrium experiments give a value of 498,000 for the molecular weight of native filamin. From these data a frictional ratio of 2.32 has been calculated. On sodium dodecyl sulfate gel electrophoresis, filamin migrates as a single protein band with an estimated molecular weight of 240,000. Filamin is a soluble protein and under a variety of conditions tested does not by itself form filaments or precipitate from solution. However, filamin binds to rabbit skeletal muscle F-actin, and the complex is readily sedimented by centrifugation to yield a gelatinous pellet containing actin and filamin. These results indicate that filamin is a dimeric protein with a moderate degree of asymmetry that binds to actin. The results also suggest that the distribution of filamin in cells is derived from its interaction with polymerized actin.

摘要

细丝蛋白是最近在鸡胗中发现的一种蛋白质（王，K.，阿什，F.，和辛格，S. J.（1975年）《美国国家科学院院刊》72，4483 - 4486），现已被提纯，不含其他成分，并对其分子特性进行了研究。细丝蛋白的沉降常数（S020,w）为8.86 S，比容为0.734 ml/g。沉降平衡实验得出天然细丝蛋白的分子量为498,000。根据这些数据计算出摩擦比为2.32。在十二烷基硫酸钠凝胶电泳中，细丝蛋白作为一条单一的蛋白带迁移，估计分子量为240,000。细丝蛋白是一种可溶性蛋白，在各种测试条件下，它本身不会形成细丝或从溶液中沉淀出来。然而，细丝蛋白能与兔骨骼肌F - 肌动蛋白结合，并且该复合物很容易通过离心沉淀，产生一个含有肌动蛋白和细丝蛋白的凝胶状沉淀。这些结果表明细丝蛋白是一种具有中等不对称程度的二聚体蛋白，能与肌动蛋白结合。结果还表明细丝蛋白在细胞中的分布源于它与聚合肌动蛋白的相互作用。