| Literature DB >> 32529363 |
Frances Joan D Alvarez1,2, Peijun Zhang3,4,5.
Abstract
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. In this chapter, 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.Entities:
Keywords: Cryo-EM; Dynamin; GTPase; Helical assembly; MBP fusion; Mammalian expression; Mx protein; Negative-staining EM; Purification
Mesh:
Substances:
Year: 2020 PMID: 32529363 PMCID: PMC8164019 DOI: 10.1007/978-1-0716-0676-6_5
Source DB: PubMed Journal: Methods Mol Biol ISSN: 1064-3745