Ngo, K. H., Lattmann, S., Anindita, P. D., Liew, C. W., Harris, R. S., Luo, D., & Kang, C. (2026). An inactive Zika NS2B-NS3pro protease construct for investigating allosteric inhibitors. Journal of Structural Biology, 218(2), 108314. https://doi.org/10.1016/j.jsb.2026.108314
Abstract:
The Zika virus protease, composed of the cofactor region from NS2B and the N-terminal region of NS3, plays a critical role in viral polyprotein maturation and represents an attractive therapeutic target. However, developing small-molecule inhibitors for its highly hydrophilic active site remains challenging, highlighting the importance of pursuing allosteric inhibition strategies. In this study, we engineered an NS2B-NS3 protease containing an 18-residue NS2B sequence linked to the N-terminal region of NS3 via a glycine-rich linker. We determined its crystal structure and obtained the solution NMR spectrum with backbone resonance assigned. This new construct was used in fragment screening and two new fragments were identified. This design excludes the C-terminal part of NS2B cofactor region, whose conformation is influenced by substrate or inhibitor binding, making the construct particularly valuable for screening and characterizing allosteric inhibitors.
License type:
Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
Funding Info:
This research / project is supported by the Singapore Ministry of Education - Academic Research Fund
Grant Reference no. : MOE-T2EP30124-0023
This research / project is supported by the Singapore-International Synchrotron Access Programme (SG-ISAP) - Singapore-International Synchrotron Access Programme (SG-ISAP)
Grant Reference no. : AS241/MXCAP20846/21617a
This research / project is supported by the United States National Institute of Allergy and Infectious Disease (NIAID) - United States National Institute of Allergy and Infectious Disease (NIAID)
Grant Reference no. : U19-AI171954