Abstract
Norovirus capsids are icosahedral particles composed of 90 dimers of the major capsid protein VP1. The C-terminus of the VP1 proteins forms a protruding (P)-domain, mediating receptor attachment, and providing a target for neutralizing antibodies. NMR and native mass spectrometry directly detect P-domain monomers in solution for murine (MNV) but not for human norovirus (HuNoV). We report that the binding of glycochenodeoxycholic acid (GCDCA) stabilizes MNV-1 P-domain dimers (P-dimers) and induces long-range NMR chemical shift perturbations (CSPs) within loops involved in antibody and receptor binding, likely reflecting corresponding conformational changes. Global line shape analysis of monomer and dimer cross-peaks in concentration-dependent methyl TROSY NMR spectra yields a dissociation rate constant koff of about 1 s−1 for MNV-1 P-dimers. For structurally closely related HuNoV GII.4 Saga P-dimers a value of about 10−6s−1 is obtained from ion-exchange chromatography, suggesting essential differences in the role of GCDCA as a cofactor for MNV and HuNoV infection.
| Original language | English |
|---|---|
| Article number | 563 |
| Journal | Communications Biology |
| Volume | 5 |
| Issue number | 1 |
| Pages (from-to) | 563 |
| DOIs | |
| Publication status | Published - 09.06.2022 |
Funding
This research was funded by the Deutsche Forschungsgemeinschaft (DFG) via grants Pe494/12-2 (T.P.) and TA1093-2 (S.T.) within the research unit FOR2327 (ViroCarb). T.P. thanks the State of Schleswig-Holstein for supplying the NMR infrastructure (European Funds for Regional Development, LPW-E/1.1.2/857). C.F. thanks the Studienstiftung des deutschen Volkes for a fellowship. J.D. and C.U. acknowledge funding from FOR2327 ViroCarb (UE 183/1–2). C.U. and L.T. acknowledge funding from the EU Horizon 2020 project VIRUSCAN 731868. The Leibniz Institute for Experimental Virology (HPI) is supported by the Free and Hanseatic City of Hamburg and the Federal Ministry of Health. T.J.S. acknowledges funding from the NIH, grant 1R01-AI141465. We thank Cristiane Wobus at the University of Michigan (Michigan, USA) for the generous gift of the hybridoma cells 2D3, 4F9, and A6.2 and Ian Goodfellow (Cambridge University, United Kingdom) for the cDNA clone “pT7MNV3’RZ” for the optimized RNA-based reverse-genetics system. We would like to thank Thomas Krey, Institute of Biochemistry, University of Lübeck, for giving us access to the SEC-MALS apparatus. This research was funded by the Deutsche Forschungsgemeinschaft (DFG) via grants Pe494/12-2 (T.P.) and TA1093-2 (S.T.) within the research unit FOR2327 (ViroCarb). T.P. thanks the State of Schleswig-Holstein for supplying the NMR infrastructure (European Funds for Regional Development, LPW-E/1.1.2/857). C.F. thanks the Studienstiftung des deutschen Volkes for a fellowship. J.D. and C.U. acknowledge funding from FOR2327 ViroCarb (UE 183/1–2). C.U. and L.T. acknowledge funding from the EU Horizon 2020 project VIRUSCAN 731868. The Leibniz Institute for Experimental Virology (HPI) is supported by the Free and Hanseatic City of Hamburg and the Federal Ministry of Health. T.J.S. acknowledges funding from the NIH, grant 1R01-AI141465. We thank Cristiane Wobus at the University of Michigan (Michigan, USA) for the generous gift of the hybridoma cells 2D3, 4F9, and A6.2 and Ian Goodfellow (Cambridge University, United Kingdom) for the cDNA clone “pT7MNV3’RZ” for the optimized RNA-based reverse-genetics system. We would like to thank Thomas Krey, Institute of Biochemistry, University of Lübeck, for giving us access to the SEC-MALS apparatus.
Research Areas and Centers
- Centers: Center for Structural and Cell Biology (CSCM/ZMSZ)
- Academic Focus: Center for Infection and Inflammation Research (ZIEL)
DFG Research Classification Scheme
- 2.21-04 Virology
