Structure and activity of human TMPRSS2 protease implicated in SARS-CoV-2 activation

  • Nat Chem Biol. 2022 Sep;18(9):963-971. doi: 10.1038/s41589-022-01059-7.
Bryan J Fraser   #  1  2 ,  Serap Beldar   #  3 ,  Almagul Seitova  3 ,  Ashley Hutchinson  3 ,  Dhiraj Mannar  4 ,  Yanjun Li  3 ,  Daniel Kwon  1  2 ,  Ruiyan Tan  1 ,  Ryan P Wilson  1 ,  Karoline Leopold  4 ,  Sriram Subramaniam  4 ,  Levon Halabelian  5  6 ,  Cheryl H Arrowsmith  7  8  9 ,  François Bénard  10  11
Affiliations
  • 1. Department of Molecular Oncology, British Columbia Cancer Research Institute, Vancouver, British Columbia, Canada.
  • 2. Department of Radiology, University of British Columbia, Vancouver, British Columbia, Canada.
  • 3. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
  • 4. Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia, Canada.
  • 5. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada. [email protected].
  • 6. Department of Pharmacology and Toxicology, University of Toronto, Toronto, Ontario, Canada. [email protected].
  • 7. Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada. [email protected].
  • 8. Princess Margaret Cancer Centre, Toronto, Ontario, Canada. [email protected].
  • 9. Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada. [email protected].
  • 10. Department of Molecular Oncology, British Columbia Cancer Research Institute, Vancouver, British Columbia, Canada. [email protected].
  • 11. Department of Radiology, University of British Columbia, Vancouver, British Columbia, Canada. [email protected].
  • # Contributed equally.
PMID: 35676539 DOI: 10.1038/s41589-022-01059-7
Abstract

Transmembrane protease, serine 2 (TMPRSS2) has been identified as key host cell factor for viral entry and pathogenesis of SARS-CoV-2. Specifically, TMPRSS2 proteolytically processes the SARS-CoV-2 Spike (S) protein, enabling virus-host membrane fusion and Infection of the airways. We present here a recombinant production strategy for enzymatically active TMPRSS2 and characterization of its matured proteolytic activity, as well as its 1.95 Å X-ray cocrystal structure with the synthetic protease inhibitor nafamostat. Our study provides a structural basis for the potent but nonspecific inhibition by nafamostat and identifies distinguishing features of the TMPRSS2 substrate binding pocket that explain specificity. TMPRSS2 cleaved SARS-CoV-2 S Protein at multiple sites, including the canonical S1/S2 cleavage site. We ranked the potency of clinical Protease Inhibitors with half-maximal inhibitory concentrations ranging from 1.4 nM to 120 µM and determined inhibitor mechanisms of action, providing the groundwork for drug development efforts to selectively inhibit TMPRSS2.

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