Co-refolding of a functional complex of Dengue NS3 protease and NS2B co-factor domain and backbone resonance assignment by solution NMR

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

SLU Authors

• Agback, Tatiana

  • Department of Molecular Sciences, Swedish University of Agricultural Sciences
    

• Agback, Peter

  • Department of Molecular Sciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Molecular Biology
Biochemistry

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