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.

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Published
2024-08-15
DOI
https://doi.org/10.17504/protocols.io.e6nvw1qb2lmk/v2
Primary Topic
Cell Image Analysis Techniques
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article
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0.00

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article

Crystallisation protocol for SARS-CoV-2 nsp3 macrodomain in P43 v2

Charles W.E. Tomlinson, L. Koekemoer, J.C. Aschenbrenner, D. Fearon et al.
Cell Image Analysis Techniques
article

Crystallisation protocol for SARS-CoV-2 nsp3 macrodomain in P43 v2

Charles W.E. Tomlinson, L. Koekemoer, J.C. Aschenbrenner, D. Fearon, Peter Marples
article en

Abstract

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.

Discovery Centre (CA)
Diamond Light Source, National Institutes of Health, National Institute of Allergy and Infectious Diseases
Openalex Percentile: Top 83%
Cell Image Analysis Techniques
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Crystallisation protocol for SARS-CoV-2 nsp3 macrodomain in P43 v2 — Charles W.E. Tomlinson, L. Koekemoer, et al. · (2024) | TGRS Research Map | TGRS