The effect of the hexahistidine-tag in the oligomerization of HSC70 constructs
The effect of the hexahistidine-tag in the oligomerization of HSC70 constructs
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DOI：
10.1016/j.jchromb.2006.07.031
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发表时间：
2006-12-05
影响因子：
3
通讯作者：
Ladjimi, Moncef
中科院分区：
文献类型：
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作者：
Amor-Mahjoub, Mouna;Suppini, Jean-Philippe;Ladjimi, Moncef
六聚组氨酸是一种融合标签，用于通过固定化金属离子亲和色谱（IMAC）分离蛋白质。在本研究中，我们已经纯化和分析了两个构建的热休克蛋白HSC 70在存在或不存在His标签（C30 WT-His（+）/C30 WT和C30 Delta L-His（+）/C30 Delta L）。通过尺寸排阻色谱（SEC）和分析性超离心（Au）分析构建体的寡聚化性质。SEC分析的结果表明，与C30 Delta L和C30 WT各自的未标记形式相比，His标签促进C30 Delta L-His（+）的二聚化，但对C30 WT-His（+）的洗脱曲线没有影响。这些观察结果也得到了Au分析的证实，这表明在His标签存在的情况下，C30 Delta L稳定为二聚体形式。这些结果强调，需要删除His标签之前，一些重组蛋白的结构表征。(c)2006 Elsevier B. V.保留所有权利。
The hexahistidine is a fusion tag used for the isolation of proteins via an immobilized metal-ion affinity chromatography (IMAC). In the present study, we have purified and analyzed two constructs of the heat shock protein HSC70 in the presence or the absence of the His-tag (C30WT-His(+)/C30WT and C30 Delta L-His(+)/C30 Delta L). The oligomerization properties of the constructs were analyzed by size exclusion chromatography (SEC) and analytical ultracentrifugation (AU). Results from SEC analysis indicated that the His-tag promotes the dimerization of C30 Delta L-His(+) but has no effect on the elution profile of C30WT-His(+), compared to their respective untagged forms C30 Delta L and C30WT. These observations were also confirmed by AU analysis which indicates that C30 Delta L is stabilized in the dimeric form in the presence of the His-tag. These results emphasize the need to remove the His-tag before structural characterization of some recombinant proteins. (c) 2006 Elsevier B.V. All rights reserved.