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Updated: Dec 18, 2025
Expression, Detergent Solubilization, and Purification of a Membrane Transporter, the MexB Multidrug Resistance Protein
Published on: December 3, 2010
Purification and Characterization of MxB
Frances Joan D Alvarez1,2, Peijun Zhang3,4,5
1Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
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.
Insights
Researchers developed a new method to purify the MxB protein, a key antiviral dynamin-like GTPase. This breakthrough allows for better study of its function in restricting dangerous viruses.
Area of Science:
- Virology
- Molecular Biology
- Protein Biochemistry
Background:
- MxB (myxovirus resistance protein B), also known as Mx2, is an interferon-induced GTPase crucial for restricting viral infections.
- The protein's intrinsically disordered N-terminal region and self-oligomerization properties have hindered previous purification efforts using standard bacterial expression systems.
Purpose of the Study:
- To establish a reliable method for expressing and purifying full-length wild-type MxB protein.
- To enable detailed characterization of MxB's GTPase activity and oligomerization functions.
Main Methods:
- Utilized suspension-adapted mammalian cells for protein expression.
- Developed a specific purification protocol for full-length MxB.
- Established assays to measure GTPase activity.
- Developed methods to analyze protein oligomerization.
Main Results:
- Successfully purified full-length wild-type MxB protein to homogeneity.
- Characterized the GTPase activity of the purified MxB protein.
- Demonstrated the oligomerization properties of MxB in vitro.
Conclusions:
- The described expression and purification procedure overcomes previous limitations, providing pure MxB for functional studies.
- This methodology facilitates in-depth investigation of MxB's antiviral mechanisms at a molecular level.
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