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Expression, purification of Zika virus membrane protein-NS2B in detergent micelles for NMR studies.
- Source :
-
Protein Expression & Purification . Feb2019, Vol. 154, p1-6. 6p. - Publication Year :
- 2019
-
Abstract
- 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 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. Highlights • Full length NS2B of Zika virus was purified. • NS2B in LMPG micelles exhibited dispersed NMR signals. • Structure and dynamics of NS2B were explored. • Free NS2B contains a long unstructured region. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 10465928
- Volume :
- 154
- Database :
- Academic Search Index
- Journal :
- Protein Expression & Purification
- Publication Type :
- Academic Journal
- Accession number :
- 133044248
- Full Text :
- https://doi.org/10.1016/j.pep.2018.09.013