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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Goujon C, Moncorge O, Bauby H, Doyle T, Ward CC, Schaller T, Hue S, Barclay WS, Schulz R, Malim MH (2013) Human MX2 is an interferon-induced post-entry inhibitor of HIV-1 infection. Nature 502(7472):559–562
Kane M, Yadav SS, Bitzegeio J, Kutluay SB, Zang T, Wilson SJ, Schoggins JW, Rice CM, Yamashita M, Hatziioannou T, Bieniasz PD (2013) MX2 is an interferon-induced inhibitor of HIV-1 infection. Nature 502(7472):563–566
Liu Z, Pan Q, Ding S, Qian J, Xu F, Zhou J, Cen S, Guo F, Liang C (2013) The interferon-inducible MxB protein inhibits HIV-1 infection. Cell Host Microbe 14(4):398–410
Schilling M, Bulli L, Weigang S, Graf L, Naumann S, Patzina C, Wagner V, Bauersfeld L, Goujon C, Hengel H, Halenius A, Ruzsics Z, Schaller T, Kochs G (2018) Human MxB protein is a Pan-herpesvirus restriction factor. J Virol 92(17):e01056–e01018
Jaguva Vasudevan AA, Bahr A, Grothmann R, Singer A, Haussinger D, Zimmermann A, Munk C (2018) MXB inhibits murine cytomegalovirus. Virology 522:158–167
Meier K, Jaguva Vasudevan AA, Zhang Z, Bahr A, Kochs G, Haussinger D, Munk C (2018) Equine MX2 is a restriction factor of equine infectious anemia virus (EIAV). Virology 523:52–63
Wang H, Bai J, Fan B, Li Y, Zhang Q, Jiang P (2016) The interferon-induced Mx2 inhibits porcine reproductive and respiratory syndrome virus replication. J Interferon Cytokine Res 36(2):129–139
Alvarez FJD, He S, Perilla JR, Jang S, Schulten K, Engelman AN, Scheres SHW, Zhang P (2017) CryoEM structure of MxB reveals a novel oligomerization interface critical for HIV restriction. Sci Adv 3(9):e1701264
Fribourgh JL, Nguyen HC, Matreyek KA, Alvarez FJD, Summers BJ, Dewdney TG, Aiken C, Zhang P, Engelman A, Xiong Y (2014) Structural insight into HIV-1 restriction by MxB. Cell Host Microbe 16(5):627–638
Haller O, Staeheli P, Schwemmle M, Kochs G (2015) Mx GTPases: dynamin-like antiviral machines of innate immunity. Trends Microbiol 23(3):154–163
Xu B, Kong J, Wang X, Wei W, Xie W, Yu XF (2015) Structural insight into the assembly of human anti-HIV dynamin-like protein MxB/Mx2. Biochem Biophys Res Commun 456(1):197–201
Ramachandran R, Schmid SL (2018) The dynamin superfamily. Curr Biol 28(8):R411–R416
Ingerman E, Nunnari J (2005) A continuous, regenerative coupled GTPase assay for dynamin-related proteins. Methods Enzymol 404:611–619
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Alvarez, F.J.D., Zhang, P. (2020). Purification and Characterization of MxB. In: Ramachandran, R. (eds) Dynamin Superfamily GTPases. Methods in Molecular Biology, vol 2159. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0676-6_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0676-6_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0675-9
Online ISBN: 978-1-0716-0676-6
eBook Packages: Springer Protocols