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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.

Original publication

DOI

10.1007/978-1-0716-0676-6_5

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2020

Volume

2159

Pages

55 - 65

Keywords

Cryo-EM, Dynamin, GTPase, Helical assembly, MBP fusion, Mammalian expression, Mx protein, Negative-staining EM, Purification