The tem-per-ature-dependent conformational ensemble of SARS-CoV-2 main protease (Mpro).

The COVID-19 pandemic, instigated by the SARS-CoV-2 coronavirus, continues to plague the globe. The SARS-CoV-2 main protease, or Mpro, is a promising target for the development of novel antiviral therapeutics. Previous X-ray crystal structures of Mpro were obtained at cryogenic tem-per-ature or room tem-per-ature only. Here we report a series of high-resolution crystal structures of unliganded Mpro across multiple tem-per-atures from cryogenic to physiological, and another at high humidity. We inter-rogate these data sets with parsimonious multiconformer models, multi-copy ensemble models, and isomorphous difference density maps. Our analysis reveals a perturbation-dependent conformational landscape for Mpro, including a mobile zinc ion inter-leaved between the catalytic dyad, mercurial conformational heterogeneity at various sites including a key substrate-binding loop, and a far-reaching intra-molecular network bridging the active site and dimer inter-face. Our results may inspire new strategies for antiviral drug development to aid preparation for future coronavirus pandemics.

The tem­per­ature-dependent conformational ensemble of SARS-CoV-2 main protease (Mpro)

X-ray crystallography at variable tem­per­ature for SARS-CoV-2 Mpro reveals a com­plex conformational landscape, including a mobile metal at the catalytic dyad, mercurial conformational heterogeneity at various sites, and an intra­molecular network bridging the active site and dimer inter­face. The COVID-19 pandemic, instigated by the SARS-CoV-2 coronavirus, continues to plague the globe. The SARS-CoV-2 main protease, or Mpro, is a promising target for the development of novel antiviral therapeutics. Previous X-ray crystal structures of Mpro were obtained at cryogenic tem­per­ature or room tem­per­ature only. Here we report a series of high-resolution crystal structures of unliganded Mpro across multiple tem­per­atures from cryogenic to physiological, and another at high humidity. We inter­rogate these data sets with parsimonious multiconformer models, multi-copy ensemble models, and isomorphous difference density maps. Our analysis reveals a perturbation-dependent conformational landscape for Mpro, including a mobile zinc ion inter­leaved between the catalytic dyad, mercurial conformational heterogeneity at various sites including a key substrate-binding loop, and a far-reaching intra­molecular network bridging the active site and dimer inter­face. Our results may inspire new strategies for antiviral drug development to aid preparation for future coronavirus pandemics.