Feramisco J R, Burridge K
J Biol Chem. 1980 Feb 10;255(3):1194-9.
Brief, low ionic strength extraction of chicken gizzard at 37 degrees C yields a solution containing a limited number of proteins including alpha-actinin, filamin, actin, desmin, and a 130,000-dalton polypeptide. The proteins are then fractionated by Mg2+- and (NH4)2SO4-induced precipitations and by ion exchange and gel filtration column chromatography to give rise to highly purified preparations of alpha-actinin, filamin, and a 130,000-dalton protein. The alpha-actinin and filamin isolated by this scheme are "native" based upon their S20,w values and their ability to bind to F-actin. These procedures, with minor modification, can be used for the purification of alpha-actinin from skeletal muscle and non-muscle tissues as well as for the purification of filamin from non-muscle tissue.
在37摄氏度下对鸡胗进行简短的低离子强度提取,可得到一种溶液,其中含有数量有限的蛋白质,包括α - 辅肌动蛋白、细丝蛋白、肌动蛋白、结蛋白和一种130,000道尔顿的多肽。然后通过Mg2 +和硫酸铵诱导沉淀以及离子交换和凝胶过滤柱色谱对这些蛋白质进行分级分离,以得到高度纯化的α - 辅肌动蛋白、细丝蛋白和130,000道尔顿蛋白质制剂。通过该方案分离得到的α - 辅肌动蛋白和细丝蛋白基于它们的S20,w值以及与F - 肌动蛋白结合的能力是“天然的”。这些程序稍作修改后,可用于从骨骼肌和非肌肉组织中纯化α - 辅肌动蛋白,以及从非肌肉组织中纯化细丝蛋白。