在非离子解离剂水合氯醛存在下的离子交换色谱法。应用于一种膜蛋白，牛心细胞色素c氧化酶。
Ion-exchange chromatography in the presence of the non-ionic dissociating agent chloral hydrate. Application to a membrane protein, bovine heart cytochrome c oxidase.
作者信息
Griffin D C, Landon M
出版信息
Biochem J. 1982 Jan 1;201(1):227-31. doi: 10.1042/bj2010227.
In previous work we have shown that aq. 100% (w/v) chloral hydrate (2,2,2-trichloroethane-1,1-diol) is a potent non-ionic protein dissociating agent. We have employed it in systems of polyacrylamide-gel electrophoresis and have demonstrated the presence of 15 components in a preparation of bovine heart cytochrome c oxidase [Griffin & Landon (1981) Biochem. J. 197, 333-344]. Here we describe the use of solutions containing aq. 100% (w/v) chloral hydrate in the ion-exchange column chromatographic separation on CM-cellulose of the alpha- and beta-chains of human haemoglobin, which we have employed as a model protein of known structure. We also describe the use of similar procedures in order to fractionate the polypeptide components of bovine heart cytochrome c oxidase. An effective separation has been obtained and we suggest that chloral hydrate-containing solutions could have general application in the ion-exchange-chromatographic analysis of membrane proteins, a procedure that has had restricted use owing to the inadequacy of non-ionic dissociating agents available previously.
在先前的工作中，我们已表明，100%（w/v）水合氯醛（2,2,2-三氯乙烷-1,1-二醇）是一种有效的非离子型蛋白质解离剂。我们已将其用于聚丙烯酰胺凝胶电泳体系，并已证实在牛心脏细胞色素c氧化酶制剂中存在15种成分[格里芬和兰登（1981年），《生物化学杂志》197, 333 - 344]。在此，我们描述了含100%（w/v）水合氯醛溶液在人血红蛋白α链和β链于CM - 纤维素上进行离子交换柱色谱分离中的应用，我们将人血红蛋白用作已知结构的模型蛋白质。我们还描述了使用类似方法来分离牛心脏细胞色素c氧化酶的多肽成分。已实现了有效的分离，并且我们认为含氯醛水合物的溶液在膜蛋白的离子交换色谱分析中可能具有广泛应用，此前由于可用的非离子解离剂不足，该方法的应用受到限制。
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