Journal Article

PUBDB-2023-05575

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png SARS-CoV-2 Mpro responds to oxidation by forming disulfide and NOS/SONOS bonds

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

2024
Nature Publishing Group UK [London]

This record in other databases:        

Please use a persistent id in citations: doi:10.1038/s41467-024-48109-3  doi:10.3204/PUBDB-2023-05575

Abstract: The main protease (M$^{pro}$) of SARS-CoV-2 is critical for viral function and a key drug target. Mpro 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.

---

Contributing Institute(s):

1. FS-CFEL-1 (Group Leader: Henry Chapman) (CFEL-I)
2. FS-CFEL-1 Fachgruppe PBIO (FS-CFEL-1-PBIO)
3. FS-CFEL-1 Fachgruppe BMX (FS-CFEL-1-BMX)
4. SPB/SFX (XFEL_E1_SPB/SFX)

Research Program(s):

1. 633 - Life Sciences – Building Blocks of Life: Structure and Function (POF4-633) (POF4-633)
2. 6G3 - PETRA III (DESY) (POF4-6G3) (POF4-6G3)
3. InternLabs-0011 - HIR3X - Helmholtz International Laboratory on Reliability, Repetition, Results at the most advanced X-ray Sources (2020_InternLabs-0011) (2020_InternLabs-0011)
4. FISCOV - FISCOV Helmholtz large research infrastructures in the fight against epidemic outbreaks (FISCOV) (FISCOV)
5. DFG project G:(GEPRIS)390715994 - EXC 2056: CUI: Advanced Imaging of Matter (390715994) (390715994)
6. DFG project G:(GEPRIS)194651731 - EXC 1074: Hamburger Zentrum für ultraschnelle Beobachtung (CUI): Struktur, Dynamik und Kontrolle von Materie auf atomarer Skala (194651731) (194651731)

Experiment(s):

1. PETRA Beamline P11 (PETRA III)
2. SPB: Single Particles, clusters & Biomolecules (SASE1)

Appears in the scientific report 2024

---

Database coverage:
; ; ; ; Article Processing Charges ; BIOSIS Previews ; Biological Abstracts ; Clarivate Analytics Master Journal List ; Current Contents - Agriculture, Biology and Environmental Sciences ; Current Contents - Life Sciences ; Current Contents - Physical, Chemical and Earth Sciences ; DOAJ Seal ; Essential Science Indicators ; Fees ; IF >= 15 ; JCR ; SCOPUS ; Science Citation Index Expanded ; Web of Science Core Collection ; Zoological Record

---