Elizabeth Yihui Ng, Ying Ru Loh, Yan Li, Qingxin Li, CongBao Kang, Expression, purification of Zika virus membrane protein-NS2B in detergent micelles for NMR studies, Protein Expression and Purification, Volume 154, 2019, Pages 1-6, ISSN 1046-5928, https://doi.org/10.1016/j.pep.2018.09.013.
Abstract:
The Zika virus (ZIKV) genome encodes a polyprotein that can be post-translationally processed into functional viral proteins. The viral protease is indispensable in the maturation of viral proteins. The Zika protease comprises of two components crucial for catalysis. The N-terminal region of NS3 contains the catalytic triad and approximately 40 amino acids of NS2B that are essential for folding and protease activity. NS2B is a membrane protein with transmembrane domains that are critical for the localization of NS3 to the membrane. In this study, we expressed and purified full-length NS2B from ZIKV in E. coli. Purified NS2B was then reconstituted into lyso-myristoyl phosphatidylglycerol (LMPG) micelles. It was found that compared to wild type NS2B, NS2B C11S mutation in LMPG exhibited dispersed cross peaks in the 1H-15N-HSQC spectrum, thereby suggesting the feasibility for structural characterization using solution NMR spectroscopy.
License type:
http://creativecommons.org/licenses/by-nc-nd/4.0/
Funding Info:
Agency for Science, Technology and Research (A*STAR) JCO grant (1431AFG102/1331A028). Science and Engineering Research Council (SERC) of A*STAR of Singapore (SERC grant number: 1526004161)