tetano
Editor, Senior Moderator
Nat Commun
. 2024 May 7;15(1):3827.
doi: 10.1038/s41467-024-48109-3. SARS-CoV-2 M[SUP]pro[/SUP] responds to oxidation by forming disulfide and NOS/SONOS bonds
Patrick Y A Reinke[SUP] #[/SUP][SUP] 1 [/SUP], Robin Schubert[SUP] #[/SUP][SUP] 2 [/SUP], Dominik Oberthür[SUP] 1 [/SUP], Marina Galchenkova[SUP] 1 [/SUP], Aida Rahmani Mashhour[SUP] 1 [/SUP], Sebastian Günther[SUP] 1 [/SUP], Anaïs Chretien[SUP] 2 [/SUP], Adam Round[SUP] 2 [/SUP], Brandon Charles Seychell[SUP] 3 [/SUP], Brenna Norton-Baker[SUP] 4 5 [/SUP], Chan Kim[SUP] 2 [/SUP], Christina Schmidt[SUP] 2 [/SUP], Faisal H M Koua[SUP] 2 [/SUP], Alexandra Tolstikova[SUP] 1 [/SUP], Wiebke Ewert[SUP] 1 [/SUP], Gisel Esperanza Peña Murillo[SUP] 1 6 [/SUP], Grant Mills[SUP] 2 [/SUP], Henry Kirkwood[SUP] 2 [/SUP], Hévila Brognaro[SUP] 7 [/SUP], Huijong Han[SUP] 2 [/SUP], Jayanath Koliyadu[SUP] 2 [/SUP], Joachim Schulz[SUP] 2 [/SUP], Johan Bielecki[SUP] 2 [/SUP], Julia Lieske[SUP] 1 [/SUP], Julia Maracke[SUP] 1 [/SUP], Juraj Knoska[SUP] 1 6 [/SUP], Kristina Lorenzen[SUP] 2 [/SUP], Lea Brings[SUP] 2 [/SUP], Marcin Sikorski[SUP] 2 [/SUP], Marco Kloos[SUP] 2 [/SUP], Mohammad Vakili[SUP] 1 2 [/SUP], Patrik Vagovic[SUP] 1 2 [/SUP], Philipp Middendorf[SUP] 1 [/SUP], Raphael de Wijn[SUP] 2 [/SUP], Richard Bean[SUP] 2 [/SUP], Romain Letrun[SUP] 2 [/SUP], Seonghyun Han[SUP] 2 8 [/SUP], Sven Falke[SUP] 1 [/SUP], Tian Geng[SUP] 9 [/SUP], Tokushi Sato[SUP] 2 [/SUP], Vasundara Srinivasan[SUP] 7 [/SUP], Yoonhee Kim[SUP] 2 [/SUP], Oleksandr M Yefanov[SUP] 1 [/SUP], Luca Gelisio[SUP] 2 [/SUP], Tobias Beck[SUP] 3 10 [/SUP], Andrew S Doré[SUP] 9 11 [/SUP], Adrian P Mancuso[SUP] 2 12 13 [/SUP], Christian Betzel[SUP] 7 10 [/SUP], Saša Bajt[SUP] 1 10 [/SUP], Lars Redecke[SUP] 14 15 [/SUP], Henry N Chapman[SUP] 1 6 10 [/SUP], Alke Meents[SUP] 1 [/SUP], Dušan Turk[SUP] 16 17 [/SUP], Winfried Hinrichs[SUP] 18 [/SUP], Thomas J Lane[SUP] 19 20 21 [/SUP]
Affiliations
The main protease (M[SUP]pro[/SUP]) of SARS-CoV-2 is critical for viral function and a key drug target. M[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] and the crystallization conditions necessary to study this structurally.
. 2024 May 7;15(1):3827.
doi: 10.1038/s41467-024-48109-3. SARS-CoV-2 M[SUP]pro[/SUP] responds to oxidation by forming disulfide and NOS/SONOS bonds
Patrick Y A Reinke[SUP] #[/SUP][SUP] 1 [/SUP], Robin Schubert[SUP] #[/SUP][SUP] 2 [/SUP], Dominik Oberthür[SUP] 1 [/SUP], Marina Galchenkova[SUP] 1 [/SUP], Aida Rahmani Mashhour[SUP] 1 [/SUP], Sebastian Günther[SUP] 1 [/SUP], Anaïs Chretien[SUP] 2 [/SUP], Adam Round[SUP] 2 [/SUP], Brandon Charles Seychell[SUP] 3 [/SUP], Brenna Norton-Baker[SUP] 4 5 [/SUP], Chan Kim[SUP] 2 [/SUP], Christina Schmidt[SUP] 2 [/SUP], Faisal H M Koua[SUP] 2 [/SUP], Alexandra Tolstikova[SUP] 1 [/SUP], Wiebke Ewert[SUP] 1 [/SUP], Gisel Esperanza Peña Murillo[SUP] 1 6 [/SUP], Grant Mills[SUP] 2 [/SUP], Henry Kirkwood[SUP] 2 [/SUP], Hévila Brognaro[SUP] 7 [/SUP], Huijong Han[SUP] 2 [/SUP], Jayanath Koliyadu[SUP] 2 [/SUP], Joachim Schulz[SUP] 2 [/SUP], Johan Bielecki[SUP] 2 [/SUP], Julia Lieske[SUP] 1 [/SUP], Julia Maracke[SUP] 1 [/SUP], Juraj Knoska[SUP] 1 6 [/SUP], Kristina Lorenzen[SUP] 2 [/SUP], Lea Brings[SUP] 2 [/SUP], Marcin Sikorski[SUP] 2 [/SUP], Marco Kloos[SUP] 2 [/SUP], Mohammad Vakili[SUP] 1 2 [/SUP], Patrik Vagovic[SUP] 1 2 [/SUP], Philipp Middendorf[SUP] 1 [/SUP], Raphael de Wijn[SUP] 2 [/SUP], Richard Bean[SUP] 2 [/SUP], Romain Letrun[SUP] 2 [/SUP], Seonghyun Han[SUP] 2 8 [/SUP], Sven Falke[SUP] 1 [/SUP], Tian Geng[SUP] 9 [/SUP], Tokushi Sato[SUP] 2 [/SUP], Vasundara Srinivasan[SUP] 7 [/SUP], Yoonhee Kim[SUP] 2 [/SUP], Oleksandr M Yefanov[SUP] 1 [/SUP], Luca Gelisio[SUP] 2 [/SUP], Tobias Beck[SUP] 3 10 [/SUP], Andrew S Doré[SUP] 9 11 [/SUP], Adrian P Mancuso[SUP] 2 12 13 [/SUP], Christian Betzel[SUP] 7 10 [/SUP], Saša Bajt[SUP] 1 10 [/SUP], Lars Redecke[SUP] 14 15 [/SUP], Henry N Chapman[SUP] 1 6 10 [/SUP], Alke Meents[SUP] 1 [/SUP], Dušan Turk[SUP] 16 17 [/SUP], Winfried Hinrichs[SUP] 18 [/SUP], Thomas J Lane[SUP] 19 20 21 [/SUP]
Affiliations
- PMID: 38714735
- PMCID: PMC11076503
- DOI: 10.1038/s41467-024-48109-3
The main protease (M[SUP]pro[/SUP]) of SARS-CoV-2 is critical for viral function and a key drug target. M[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] 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[SUP]pro[/SUP] and the crystallization conditions necessary to study this structurally.