Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography

STAR Protoc. 2022 Jan 24;3(1):101158. doi: 10.1016/j.xpro.2022.101158. eCollection 2022 Mar 18.

Abstract

The SARS-CoV-2 main protease of (Mpro) is an important target for SARS-CoV-2 related drug repurposing and development studies. Here, we describe the steps for structural characterization of SARS-CoV-2 Mpro, starting from plasmid preparation and protein purification. We detail the steps for crystallization using the sitting drop, microbatch (under oil) approach. Finally, we cover data collection and structure determination using serial femtosecond crystallography. For complete details on the use and execution of this protocol, please refer to Durdagi et al. (2021).

Keywords: Protein Biochemistry; Protein expression and purification; Structural Biology; X-ray Crystallography.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Coronavirus 3C Proteases / chemistry*
  • Coronavirus 3C Proteases / genetics
  • Crystallization
  • Crystallography, X-Ray
  • Humans
  • Models, Molecular*
  • SARS-CoV-2 / enzymology*

Substances

  • 3C-like proteinase, SARS-CoV-2
  • Coronavirus 3C Proteases