7PXZ

Reduced form of SARS-CoV-2 Main Protease determined by XFEL radiation

7PXZ の概要

• エントリーDOI: 10.2210/pdb7pxz/pdb
• 関連するPDBエントリー: 7PZQ
• 分子名称: 3C-like proteinase nsp5, CHLORIDE ION (3 entities in total)
• 機能のキーワード: 3c-like protease, main-protease, viral replication, polyprotein maturation, viral protein
• 由来する生物種: Severe acute respiratory syndrome coronavirus 2
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 33861.00

構造登録者

Schubert, R.,Reinke, P.,Galchenkova, M.,Oberthuer, D.,Murillo, G.E.P.,Kim, C.,Bean, R.,Turk, D.,Hinrichs, W.,Middendorf, P.,Round, A.,Schmidt, C.,Mills, G.,Kirkwood, H.,Han, H.,Koliyadu, J.,Bielecki, J.,Gelisio, L.,Sikorski, M.,Kloos, M.,Vakilii, M.,Yefanov, O.N.,Vagovic, P.,de-Wijn, R.,Letrun, R.,Guenther, S.,White, T.A.,Sato, T.,Srinivasan, V.,Kim, Y.,Chretien, A.,Han, S.,Brognaro, H.,Maracke, J.,Knoska, J.,Seychell, B.C.,Brings, L.,Norton-Baker, B.,Geng, T.,Dore, A.S.,Uetrecht, C.,Redecke, L.,Beck, T.,Lorenzen, K.,Betzel, C.,Mancuso, A.P.,Bajt, S.,Chapman, H.N.,Meents, A.,Lane, T.J. (登録日: 2021-10-08, 公開日: 2023-01-18, 最終更新日: 2024-07-31)

主引用文献

Reinke, P.Y.A.,Schubert, R.,Oberthur, D.,Galchenkova, M.,Rahmani Mashhour, A.,Gunther, S.,Chretien, A.,Round, A.,Seychell, B.C.,Norton-Baker, B.,Kim, C.,Schmidt, C.,Koua, F.H.M.,Tolstikova, A.,Ewert, W.,Pena Murillo, G.E.,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.,Dore, A.S.,Mancuso, A.P.,Betzel, C.,Bajt, S.,Redecke, L.,Chapman, H.N.,Meents, A.,Turk, D.,Hinrichs, W.,Lane, T.J.
SARS-CoV-2 M pro responds to oxidation by forming disulfide and NOS/SONOS bonds.
Nat Commun, 15:3827-3827, 2024

PubMed Abstract: The main protease (M) of SARS-CoV-2 is critical for viral function and a key drug target. M 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 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 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 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 and the crystallization conditions necessary to study this structurally.

PubMed: 38714735
DOI: 10.1038/s41467-024-48109-3

実験手法

X-RAY DIFFRACTION (1.75 Å)