학술논문
N-Terminal Finger Stabilizes the S1 Pocket for the Reversible Feline Drug GC376 in the SARS-CoV-2 M pro Dimer.
Document Type
Academic Journal
Author
Arutyunova E; Department of Biochemistry, Faculty of Medicine and Dentistry, Membrane Protein Disease Research Group, University of Alberta, Edmonton T6G 2R3, Alberta, Canada; Li Ka Shing Institute of Virology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2E1, Alberta, Canada.; Khan MB; Department of Biochemistry, Faculty of Medicine and Dentistry, Membrane Protein Disease Research Group, University of Alberta, Edmonton T6G 2R3, Alberta, Canada.; Fischer C; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; Lu J; Department of Biochemistry, Faculty of Medicine and Dentistry, Membrane Protein Disease Research Group, University of Alberta, Edmonton T6G 2R3, Alberta, Canada; Li Ka Shing Institute of Virology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2E1, Alberta, Canada.; Lamer T; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; Vuong W; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; van Belkum MJ; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; McKay RT; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; Tyrrell DL; Department of Medical Microbiology and Immunology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2E1, Alberta, Canada; Li Ka Shing Institute of Virology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2E1, Alberta, Canada.; Vederas JC; Department of Chemistry, Faculty of Science, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; Young HS; Department of Biochemistry, Faculty of Medicine and Dentistry, Membrane Protein Disease Research Group, University of Alberta, Edmonton T6G 2R3, Alberta, Canada. Electronic address: hyoung@ualberta.ca.; Lemieux MJ; Department of Biochemistry, Faculty of Medicine and Dentistry, Membrane Protein Disease Research Group, University of Alberta, Edmonton T6G 2R3, Alberta, Canada; Li Ka Shing Institute of Virology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton T6G 2E1, Alberta, Canada. Electronic address: mlemieux@ualberta.ca.
Source
Publisher: Elsevier Country of Publication: Netherlands NLM ID: 2985088R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1089-8638 (Electronic) Linking ISSN: 00222836 NLM ISO Abbreviation: J Mol Biol Subsets: MEDLINE
Subject
Language
English
Abstract
The main protease (M pro , also known as 3CL protease) of SARS-CoV-2 is a high priority drug target in the development of antivirals to combat COVID-19 infections. A feline coronavirus antiviral drug, GC376, has been shown to be effective in inhibiting the SARS-CoV-2 main protease and live virus growth. As this drug moves into clinical trials, further characterization of GC376 with the main protease of coronaviruses is required to gain insight into the drug's properties, such as reversibility and broad specificity. Reversibility is an important factor for therapeutic proteolytic inhibitors to prevent toxicity due to off-target effects. Here we demonstrate that GC376 has nanomolar K i values with the M pro from both SARS-CoV-2 and SARS-CoV strains. Restoring enzymatic activity after inhibition by GC376 demonstrates reversible binding with both proteases. In addition, the stability and thermodynamic parameters of both proteases were studied to shed light on physical chemical properties of these viral enzymes, revealing higher stability for SARS-CoV-2 M pro . The comparison of a new X-ray crystal structure of M pro from SARS-CoV complexed with GC376 reveals similar molecular mechanism of inhibition compared to SARS-CoV-2 M pro , and gives insight into the broad specificity properties of this drug. In both structures, we observe domain swapping of the N-termini in the dimer of the M pro , which facilitates coordination of the drug's P1 position. These results validate that GC376 is a drug with an off-rate suitable for clinical trials.
Competing Interests: Competing interests The authors declare no competing interests.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)
Competing Interests: Competing interests The authors declare no competing interests.
(Copyright © 2021 Elsevier Ltd. All rights reserved.)