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

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 crystalli...

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Autores principales: Ertem, Fatma Betul, Guven, Omur, Buyukdag, Cengizhan, Gocenler, Oktay, Ayan, Esra, Yuksel, Busra, Gul, Mehmet, Usta, Gozde, Cakılkaya, Barıs, Johnson, J. Austin, Dao, E. Han, Su, Zhen, Poitevin, Frederic, Yoon, Chun Hong, Kupitz, Christopher, Hayes, Brandon, Liang, Mengning, Hunter, Mark S., Batyuk, Alexander, Sierra, Raymond G., Ketawala, Gihan, Botha, Sabine, Dağ, Çağdaş, DeMirci, Hasan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784426/
https://www.ncbi.nlm.nih.gov/pubmed/35194584
http://dx.doi.org/10.1016/j.xpro.2022.101158
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author Ertem, Fatma Betul
Guven, Omur
Buyukdag, Cengizhan
Gocenler, Oktay
Ayan, Esra
Yuksel, Busra
Gul, Mehmet
Usta, Gozde
Cakılkaya, Barıs
Johnson, J. Austin
Dao, E. Han
Su, Zhen
Poitevin, Frederic
Yoon, Chun Hong
Kupitz, Christopher
Hayes, Brandon
Liang, Mengning
Hunter, Mark S.
Batyuk, Alexander
Sierra, Raymond G.
Ketawala, Gihan
Botha, Sabine
Dağ, Çağdaş
DeMirci, Hasan
author_facet Ertem, Fatma Betul
Guven, Omur
Buyukdag, Cengizhan
Gocenler, Oktay
Ayan, Esra
Yuksel, Busra
Gul, Mehmet
Usta, Gozde
Cakılkaya, Barıs
Johnson, J. Austin
Dao, E. Han
Su, Zhen
Poitevin, Frederic
Yoon, Chun Hong
Kupitz, Christopher
Hayes, Brandon
Liang, Mengning
Hunter, Mark S.
Batyuk, Alexander
Sierra, Raymond G.
Ketawala, Gihan
Botha, Sabine
Dağ, Çağdaş
DeMirci, Hasan
author_sort Ertem, Fatma Betul
collection PubMed
description 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).
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spelling pubmed-87844262022-01-24 Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography Ertem, Fatma Betul Guven, Omur Buyukdag, Cengizhan Gocenler, Oktay Ayan, Esra Yuksel, Busra Gul, Mehmet Usta, Gozde Cakılkaya, Barıs Johnson, J. Austin Dao, E. Han Su, Zhen Poitevin, Frederic Yoon, Chun Hong Kupitz, Christopher Hayes, Brandon Liang, Mengning Hunter, Mark S. Batyuk, Alexander Sierra, Raymond G. Ketawala, Gihan Botha, Sabine Dağ, Çağdaş DeMirci, Hasan STAR Protoc Protocol 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). Elsevier 2022-01-24 /pmc/articles/PMC8784426/ /pubmed/35194584 http://dx.doi.org/10.1016/j.xpro.2022.101158 Text en © 2022. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ertem, Fatma Betul
Guven, Omur
Buyukdag, Cengizhan
Gocenler, Oktay
Ayan, Esra
Yuksel, Busra
Gul, Mehmet
Usta, Gozde
Cakılkaya, Barıs
Johnson, J. Austin
Dao, E. Han
Su, Zhen
Poitevin, Frederic
Yoon, Chun Hong
Kupitz, Christopher
Hayes, Brandon
Liang, Mengning
Hunter, Mark S.
Batyuk, Alexander
Sierra, Raymond G.
Ketawala, Gihan
Botha, Sabine
Dağ, Çağdaş
DeMirci, Hasan
Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title_full Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title_fullStr Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title_full_unstemmed Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title_short Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
title_sort protocol for structure determination of sars-cov-2 main protease at near-physiological-temperature by serial femtosecond crystallography
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8784426/
https://www.ncbi.nlm.nih.gov/pubmed/35194584
http://dx.doi.org/10.1016/j.xpro.2022.101158
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