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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 in molecular biology (Clifton, N.J.)

Publication Date

01/2020

Volume

2159

Pages

55 - 65

Addresses

Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.

Keywords

Animals, Humans, GTP Phosphohydrolases, Recombinant Fusion Proteins, Chromatography, Affinity, Chromatography, Gel, Transfection, Gene Expression, Enzyme Activation, Plasmids, HEK293 Cells, Myxovirus Resistance Proteins