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

Coronavirus 3C Proteases / chemistry Crystallization Crystallography, X-Ray Humans Models, Molecular SARS-CoV-2 / enzymology
Protein Biochemistry Protein expression and purification Structural Biology X-ray Crystallography

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
18 03 2022
Historique:
entrez: 23 2 2022
pubmed: 24 2 2022
medline: 3 3 2022
Statut: epublish

Résumé

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).

Identifiants

DOI: 10.1016/j.xpro.2022.101158 PMID: 35194584 PMC: PMC8784426
pubmed: 35194584
doi: 10.1016/j.xpro.2022.101158
pii: S2666-1667(22)00038-7
pmc: PMC8784426
doi:

Substances chimiques

3C-like proteinase, SARS-CoV-2 EC 3.4.22.-
Coronavirus 3C Proteases EC 3.4.22.28

Types de publication

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

Langues

eng

Sous-ensembles de citation

IM

Pagination

101158

Subventions

Organisme : NIGMS NIH HHS
ID : P41 GM103393
Pays : United States

Informations de copyright

© 2022.

Déclaration de conflit d'intérêts

The authors declare no competing interests.

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Auteurs

Fatma Betul Ertem (FB)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Omur Guven (O)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Cengizhan Buyukdag (C)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Oktay Gocenler (O)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Esra Ayan (E)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Busra Yuksel (B)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Mehmet Gul (M)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Gozde Usta (G)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

Barıs Cakılkaya (B)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

J Austin Johnson (JA)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.

E Han Dao (EH)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Zhen Su (Z)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.
Department of Applied Physics, Stanford University, Stanford, CA, USA.

Frederic Poitevin (F)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Chun Hong Yoon (CH)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Christopher Kupitz (C)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Brandon Hayes (B)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Mengning Liang (M)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Mark S Hunter (MS)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Alexander Batyuk (A)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Raymond G Sierra (RG)

Linac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA.

Gihan Ketawala (G)

Department of Physics, Arizona State University, Tempe, AZ 85287-1504, USA.
Biodesign Center for Applied Structural Discovery, Arizona State University, Tempe, AZ 85287-5001, USA.

Sabine Botha (S)

Department of Physics, Arizona State University, Tempe, AZ 85287-1504, USA.
Biodesign Center for Applied Structural Discovery, Arizona State University, Tempe, AZ 85287-5001, USA.

Çağdaş Dağ (Ç)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.
Koc University, Nanofabrication and Nanocharacterization Center for Scientific and Technological Advanced Research (nSTAR), Istanbul, Turkey.
Koc University Isbank Center for Infectious Diseases (KUISCID), Istanbul, Turkey.

Hasan DeMirci (H)

Department of Molecular Biology and Genetics, Koc University, Istanbul, Turkey.
Koc University Isbank Center for Infectious Diseases (KUISCID), Istanbul, Turkey.
Stanford PULSE Institute, SLAC National Laboratory, Menlo Park, CA, USA.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines virales › Protéines virales non structurales › Protéases virales › Protéases virales 3C › Protéases 3C des coronavirus › Protocol for structure determination of...
questionsmedicales.fr › Phénomènes chimiques › Cristallisation › Protocol for structure determination of...
questionsmedicales.fr › Techniques d'investigation › Techniques de chimie analytique › Cristallographie aux rayons X › Protocol for structure determination of...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › Protocol for structure determination of...
questionsmedicales.fr › Techniques d'investigation › Modèles théoriques › Modèles moléculaires › Protocol for structure determination of...
questionsmedicales.fr › Virus › Virus à ARN › Virus à ARN à brin positif › Nidovirales › Coronaviridae › Coronavirus › Betacoronavirus › Virus du SRAS › SARS-CoV-2 › Protocol for structure determination of...