Agback, Tatiana
- Department of Molecular Sciences, Swedish University of Agricultural Sciences
Research article2017Peer reviewed
Co-refolding of a functional complex of Dengue NS3 protease and NS2B co-factor domain and backbone resonance assignment by solution NMR
Woestenenk, Esmeralda; Agback, Peter; Unnerstale, Sofia; Henderson, Ian; Agback, Tatiana
Abstract
A novel approach for separate expression of dengue virus NS3 protease and its NS2B cofactor domain is described in this paper. The two proteins are expressed in E.coli and purified separately and subsequently efficiently co-refolded to form a stable complex. This straightforward and robust method allows for separate isotope labeling of the two proteins, facilitating analysis by nuclear magnetic resonance (NMR) spectroscopy. Unlinked NS2B-NS3pro behaves better in NMR spectroscopy than linked NS2B-NS3pro, which has resulted in the backbone resonance assignment of the unlinked NS2B-NS3 complex bound to a peptidic boronic acid inhibitor. (C) 2017 Elsevier Inc. All rights reserved.
Keywords
Dengue virus; NS3 protease; Flavivirus protease; Refolding; Protein purification; Resonance assignment; Protein complex
Published in
Protein Expression and Purification
2017, Volume: 140, pages: 16-27
Publisher: ACADEMIC PRESS INC ELSEVIER SCIENCE
Agback, Peter
- Department of Molecular Sciences, Swedish University of Agricultural Sciences
- Department of Molecular Sciences, Swedish University of Agricultural Sciences
SLU Authors
UKÄ Subject classification
Biochemistry and Molecular Biology
Publication identifier
DOI: https://doi.org/10.1016/j.pep.2017.07.002
Permanent link to this page (URI)
https://res.slu.se/id/publ/93161