화학공학소재연구정보센터
Protein Expression and Purification, Vol.154, 1-6, 2019
Expression, purification of Zika virus membrane protein-NS2B in detergent micelles for NMR studies
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 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 lysomyristoyl phosphatidylglycerol (LMPG) micelles. It was found that compared to wild type NS2B, NS2B C11S mutation in LMPG exhibited dispersed cross peaks in the H-1- N-15-HSQC spectrum, thereby suggesting the feasibility for structural characterization using solution NMR spectroscopy.