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来自小鼠艾氏腹水瘤细胞和兔肺的钙结合蛋白形成非共价二聚体。

Calcyclin from mouse Ehrlich ascites tumor cells and rabbit lung form non-covalent dimers.

作者信息

Wojda U, Kuźnicki J

机构信息

Nencki Institute of Experimental Biology, Warsaw, Poland.

出版信息

Biochim Biophys Acta. 1994 Dec 14;1209(2):248-52. doi: 10.1016/0167-4838(94)90192-9.

Abstract

Crosslinking treatments of fresh cytosol from mouse Ehrlich ascites tumor (EAT) cells revealed the existence of calcyclin dimers which were sensitive to SDS, but not to reducing agents, which suggests the existence of non-covalent dimers. In stored EAT cell cytosol and preparations of purified calcyclin dimers were also formed by S-S bridging (covalent dimers). The S-S dimers did not bind to organomercurial Agarose and could be separated from reduced forms of calcyclin that bound to the resin. Calcyclin eluted from the resin with DTT was a mixture of monomers and non-covalent dimers as shown by crosslinking and subsequent immunoblotting. Calcyclin from rabbit lung, lacking a cysteine residue, could also be crosslinked as a dimer. It is suggested that the ability of calcyclin to form non-covalent dimers is of physiological significance.

摘要

对来自小鼠艾氏腹水瘤(EAT)细胞的新鲜胞质溶胶进行交联处理,结果显示存在对十二烷基硫酸钠(SDS)敏感但对还原剂不敏感的钙结合蛋白二聚体,这表明存在非共价二聚体。在储存的EAT细胞胞质溶胶中,通过二硫键桥连(共价二聚体)也会形成纯化的钙结合蛋白二聚体制剂。这些二硫键二聚体不与有机汞琼脂糖结合,并且可以与结合到树脂上的还原形式的钙结合蛋白分离。用二硫苏糖醇(DTT)从树脂上洗脱下来的钙结合蛋白是单体和非共价二聚体的混合物,交联和随后的免疫印迹结果表明了这一点。来自兔肺的缺乏半胱氨酸残基的钙结合蛋白也可以交联形成二聚体。有人提出,钙结合蛋白形成非共价二聚体的能力具有生理意义。

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