Purification and Characterization of MxB.
Purification and Characterization of MxB.
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DOI：
10.1007/978-1-0716-0676-6_5
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhang P
中科院分区：
文献类型：
--
作者：
Alvarez FJD;Zhang P
MXB/MX2是一种干扰素诱导的动力素样GTP酶，可限制许多危及生命的病毒。由于其N-末端区域被认为是内在无序的，以及其自我齐聚的倾向，在传统的大肠杆菌表达系统中，全长蛋白的纯化一直没有成功。在这里，我们描述了一种从悬浮适应的哺乳动物细胞中获得全长野生型MXB的表达和纯化方法。我们进一步描述了如何表征其GTPase活性和寡聚功能。
MxB/Mx2 is an interferon-induced dynamin-like GTPase, which restricts a number of life-threatening viruses. Because of its N-terminal region, predicted to be intrinsically disordered, and its propensity to self-oligomerize, purification of the full-length protein has not been successful in conventional E. coli expression systems. Here, we describe an expression and purification procedure to obtain pure full-length wild-type MxB from suspension-adapted mammalian cells. We further describe how to characterize its GTPase activity and oligomerization function.