SARS-CoV-2 Mpro responds to oxidation by forming disulfide and NOS/SONOS bonds

Reinke, P. Y. A.; Schubert, R.; Oberthür, D.; Galchenkova, M.; Mashhour, A. R.; Günther, S.; Chretien, A.; Round, A.; Seychell, B. C.; Norton-Baker, B.; Kim, C.; Schmidt, C.; Koua, F. H. M.; Tolstikova, A.; Ewert, W.; Murillo, G. E. P.; Mills, G.; Kirkwood, H.; Brognaro, H.; Han, H.; Koliyadu, J.; Schulz, J.; Bielecki, J.; Lieske, J.; Maracke, J.; Knoska, J.; Lorenzen, K.; Brings, L.; Sikorski, M.; Kloos, M.; Vakili, M.; Vagovic, P.; Middendorf, P.; de Wijn, R.; Bean, R.; Letrun, R.; Han, S.; Falke, S.; Geng, T.; Sato, T.; Srinivasan, V.; Kim, Y.; Yefanov, O. M.; Gelisio, L.; Beck, T.; Doré, A. S.; Mancuso, A. P.; Betzel, C.; Bajt, S.; Redecke, L.; Chapman, H. N.; Meents, A.; Turk, D.; Hinrichs, W.; Lane, T. J.

doi:10.1038/s41467-024-48109-3

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SARS-CoV-2 M^pro responds to oxidation by forming disulfide and NOS/SONOS bonds

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Norton-Baker, B.
Miller Group, Atomically Resolved Dynamics Department, Max Planck Institute for the Structure and Dynamics of Matter, Max Planck Society;
Department of Chemistry, University of California at Irvine;

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Citation

Reinke, P. Y. A., Schubert, R., Oberthür, D., Galchenkova, M., Mashhour, A. R., Günther, S., et al. (2024). SARS-CoV-2 M^pro responds to oxidation by forming disulfide and NOS/SONOS bonds. Nature Communications, 15(1): 3827. doi:10.1038/s41467-024-48109-3.

Cite as: https://hdl.handle.net/21.11116/0000-000F-47B9-1

Abstract

The main protease (M^pro) of SARS-CoV-2 is critical for viral function and a key drug target. M^pro is only active when reduced; turnover ceases upon oxidation but is restored by re-reduction. This suggests the system has evolved to survive periods in an oxidative environment, but the mechanism of this protection has not been confirmed. Here, we report a crystal structure of oxidized M^pro showing a disulfide bond between the active site cysteine, C145, and a distal cysteine, C117. Previous work proposed this disulfide provides the mechanism of protection from irreversible oxidation. M^pro forms an obligate homodimer, and the C117-C145 structure shows disruption of interactions bridging the dimer interface, implying a correlation between oxidation and dimerization. We confirm dimer stability is weakened in solution upon oxidation. Finally, we observe the protein’s crystallization behavior is linked to its redox state. Oxidized M^pro spontaneously forms a distinct, more loosely packed lattice. Seeding with crystals of this lattice yields a structure with an oxidation pattern incorporating one cysteine-lysine-cysteine (SONOS) and two lysine-cysteine (NOS) bridges. These structures further our understanding of the oxidative regulation of M^pro and the crystallization conditions necessary to study this structurally.