Exploring the binding of peptidic West Nile virus NS2B–NS3 protease inhibitors by NMR

Kang, CongBao; Gayen, Shovanlal; Wang, Weiling; Severin, René; Chen, Angela Shuyi; Lim, Huichang Annie; Chia, C. S. Brian; Schüller, Andreas; Doan, Danny N.P.; Poulsen, Anders; Hill, Jeffrey; Vasudevan, Subhash G.; Keller, Thomas H.

doi:10.1016/j.antiviral.2012.11.008

Cited by 33 publications

(50 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The NMR spectrum of the linked protease showed fewer cross-peaks than predicted very likely due to the conformational flexibility of this protease construct ( Fig. 2A), which was also observed in previous studies and is similar to WNV protease (8,23,27,28). Surprisingly, removal of the Gly 4 -Ser-Gly 4 linker from the constructs and coexpression of NS2B and NS3 as an unlinked protein complex greatly improved the quality of the 15 N HSQC spectra (Fig.…”

Section: Nmr Spectra Of the Purified Protease-supporting

confidence: 54%

NMR Analysis of a Novel Enzymatically Active Unlinked Dengue NS2B-NS3 Protease Complex

Kim

Gayen

Kang

et al. 2013

Journal of Biological Chemistry

Self Cite

178

View full text Add to dashboard Cite

Background: Dengue protease is a two-component protease that is important for viral replication. Results: An unlinked protease complex containing the NS2B regulatory region and the NS3 protease domain was obtained. Conclusion:The unlinked protease complex produces dispersed cross-peaks in NMR spectra and exists predominantly in a closed conformation in solution.Significance: This new construct will be a useful tool for drug discovery against the dengue virus.

show abstract

Section: Nmr Spectra Of the Purified Protease-supporting

confidence: 54%

NMR Analysis of a Novel Enzymatically Active Unlinked Dengue NS2B-NS3 Protease Complex

Kim

Gayen

Kang

et al. 2013

Journal of Biological Chemistry

Self Cite

178

View full text Add to dashboard Cite

show abstract

“…AcKR‐H contains an aldehyde moiety and is a potent inhibitor of WNV protease with an IC 50 in the nanomolar range . It also forms a tight complex with gZiPro as demonstrated by titration experiments showing that the binding is undergoing slow exchange in the NMR time regime (Fig.…”

Section: Resultsmentioning

confidence: 96%

Structural characterization of the linked NS2B‐NS3 protease of Zika virus

Phoo

Loh

et al. 2017

FEBS Letters

Self Cite

View full text Add to dashboard Cite

The Zika virus (ZIKV) NS2B-NS3 protease is an important drug target. The conventional flaviviral protease constructs used for structural studies contain the NS2B cofactor region linked to the NS3 protease domain via a glycine-rich flexible linker. Here, we examined the structural dynamics of this conventional Zika protease (gZiPro) using NMR spectroscopy. Although the glycine-rich linker in gZiPro does not alter the overall folding of the protease in solution, gZiPro is not homogenous in ion exchange chromatography. Compared to the unlinked protease construct, the artificial linker affects the chemical environment of many residues including H51 in the catalytic triad. Our study provides a direct comparison of ZIKV protease constructs with and without an artificial linker.

show abstract

“…Inhibitor designs for DENpro have used either conformation as template [14,17,19,20,23,26,[42][43][44][45][46][47][48][49][50][51][52]. For both WNVpro and DENpro, the presence of low molecular mass inhibitors greatly improves the quality of the NMR spectra [11,38,44,[53][54][55]. For WNVpro, however, the NMR experiments clearly showed that the closed conformation is predominant in solution regardless of the presence of an inhibitor [38].…”

Section: Open and Closed Conformationsmentioning

confidence: 99%

Binding mode of the activity‐modulating C‐terminal segment of NS2B to NS3 in the dengue virus NS2B–NS3 protease

Cruz

Chen

Graham³

et al. 2014

The FEBS Journal

View full text Add to dashboard Cite

The two-component dengue virus NS2B-NS3 protease (NS2B-NS3pro) is an established drug target but inhibitor design is hampered by uncertainties about its 3D structure in solution. Crystal structures reported very different conformations for the functionally important C-terminal segment of the NS2B cofactor (NS2Bc), indicating open and closed conformations in the absence and presence of inhibitors, respectively. An earlier NMR study in solution indicated that a closed state is the preferred conformation in the absence of an artificial linker engineered between NS2B and NS3pro. To obtain direct structural information on the fold of unlinked NS2B-NS3pro in solution, we tagged NS3pro with paramagnetic tags and measured pseudocontact shifts by NMR to position NS2Bc relative to NS3pro. NS2Bc was found to bind to NS3pro in the same way as reported in a previously published model and crystal structure of the closed state. The structure is destabilized, however, by high ionic strength and basic pH, showing the importance of electrostatic forces to tie NS2Bc to NS3pro. Narrow NMR signals previously thought to represent the open state are associated with protein degradation. In conclusion, the closed conformation of the NS2B-NS3 protease is the best model for structure-guided drug design.

show abstract

Exploring the binding of peptidic West Nile virus NS2B–NS3 protease inhibitors by NMR

Cited by 33 publications

References 27 publications

NMR Analysis of a Novel Enzymatically Active Unlinked Dengue NS2B-NS3 Protease Complex

NMR Analysis of a Novel Enzymatically Active Unlinked Dengue NS2B-NS3 Protease Complex

Structural characterization of the linked NS2B‐NS3 protease of Zika virus

Binding mode of the activity‐modulating C‐terminal segment of NS2B to NS3 in the dengue virus NS2B–NS3 protease

Contact Info

Product

Resources

About