Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity
Bacterial expression and purification of Interleukin-2 Tyrosine kinase: Single step separation of the chaperonin impurity
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DOI：
10.1016/j.pep.2008.04.001
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发表时间：
2008-08-01
影响因子：
1.6
通讯作者：
Andreotti, Amy H.
中科院分区：
文献类型：
--
作者：
Joseph, Raji E.;Andreotti, Amy H.
激酶的生物化学和生物物理特性需要大量纯化的蛋白质。在这里，我们报告的细菌表达和纯化的活性Itk激酶结构域（Tec家族激酶）使用ArcticExpress细胞，共表达伴侣蛋白系统Cpn 60/10从Oleispira anaitica。我们描述了一个简单的一步MgCl 2/ATP/KCl孵育程序，以去除共纯化伴侣蛋白杂质。伴侣蛋白共纯化是蛋白质纯化过程中遇到的常见问题，本文所述的简单孵育步骤完全克服了这个问题。该方法靶向伴侣蛋白系统而不是感兴趣的蛋白质，因此广泛适用于其他蛋白质靶点。(c)2008年爱思唯尔公司All rights reserved.
Biochemical and biophysical characterization of kinases requires large quantities of purified protein. Here, we report the bacterial expression and purification of active Itk kinase domain (a Tec family kinase) using ArcticExpress cells that co-express the chaperonin system Cpn60/10 from Oleispira antarctica. We describe a simple one step MgCl2/ATP/KCl incubation procedure to remove the co-purifying chaperonin impurity. Chaperonin co-purification is a common problem encountered during protein purification and the simple incubation step described here completely overcomes this problem. The approach targets the chaperonin system rather than the protein of interest and is therefore widely applicable to other protein targets. (c) 2008 Elsevier Inc. All rights reserved.