Crystallisation protocol for SARS-CoV-2 nsp3 macrodomain in P43 v2
The COVID-19 pandemic has demonstrated the need for novel therapeutic interventions and improved pandemic preparedness strategies against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This protocol details an optimized crystallization method for the SARS-CoV-2 nsp3 macrodomain, a potential drug target. Using sitting drop vapor diffusion, we describe specific buffer conditions and procedures to consistently produce high-quality crystals suitable for XChem fragment screening. The method yields crystals that diffract to an average resolution of 1.2 Å, enabling high-resolution structural studies. All structures solved during the development of tool compounds for the SARS-CoV-2 nsp3 macrodomain are deposited on the PDB (Group deposition: G_1002283). In this version we added the Addgene id and changed the results section.
Authors
- Charles W.E. Tomlinson (ORCID: https://orcid.org/0000-0002-1845-6028)
- L. Koekemoer (ORCID: https://orcid.org/0000-0001-9226-9127)
- J.C. Aschenbrenner (ORCID: https://orcid.org/0000-0002-4318-0481)
- D. Fearon (ORCID: https://orcid.org/0000-0003-3529-7863)
- Peter Marples (ORCID: https://orcid.org/0000-0002-8787-7969)
Institutions
- Discovery Centre (CA)
Publication Details
- Published
- 2024-08-15
- DOI
- https://doi.org/10.17504/protocols.io.e6nvw1qb2lmk/v2
- Primary Topic
- Cell Image Analysis Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Diamond Light Source
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases