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

Author

Ertem, Fatma Betul, primary, Guven, Omur, additional, Buyukdag, Cengizhan, additional, Gocenler, Oktay, additional, Ayan, Esra, additional, Yuksel, Busra, additional, Gul, Mehmet, additional, Usta, Gozde, additional, Cakılkaya, Barıs, additional, Johnson, J. Austin, additional, Dao, E. Han, additional, Su, Zhen, additional, Poitevin, Frederic, additional, Yoon, Chun Hong, additional, Kupitz, Christopher, additional, Hayes, Brandon, additional, Liang, Mengning, additional, Hunter, Mark S., additional, Batyuk, Alexander, additional, Sierra, Raymond G., additional, Ketawala, Gihan, additional, Botha, Sabine, additional, Dağ, Çağdaş, additional, and DeMirci, Hasan, additional

Published
2022
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2. Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography

Author

Fatma Betul Ertem, Omur Guven, Cengizhan Buyukdag, Oktay Gocenler, Esra Ayan, Busra Yuksel, Mehmet Gul, Gozde Usta, Barıs Cakılkaya, J. Austin Johnson, E. Han Dao, Zhen Su, Frederic Poitevin, Chun Hong Yoon, Christopher Kupitz, Brandon Hayes, Mengning Liang, Mark S. Hunter, Alexander Batyuk, Raymond G. Sierra, Gihan Ketawala, Sabine Botha, Çağdaş Dağ, Hasan DeMirci, Ertem, Fatma Betül, Güven, Ömür, Büyükdağ, Cengizhan, Göçenler, Oktay, Ayan, Esra, Yüksel, Büşra, Gül, Mehmet, Usta, Gözde, Çakılkaya, Barış, Johnson, J. Austin, Demirci, Hasan, Dağ, Çağdaş, Demirci, Hasan (ORCID 0000-0002-9135-5397 & YÖK ID 307350), 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, Koç Üniversitesi İş Bankası Enfeksiyon Hastalıkları Uygulama ve Araştırma Merkezi (EHAM) / Koç University İşbank Center for Infectious Diseases (KU-IS CID), Graduate School of Sciences and Engineering, College of Sciences, and Department of Molecular Biology and Genetics

Subjects
Models, Molecular, Science (General), General Immunology and Microbiology, SARS-CoV-2, General Neuroscience, fungi, Crystallography, X-Ray, General Biochemistry, Genetics and Molecular Biology, body regions, Q1-390, X-ray Crystallography, X-Ray laser, Crystallography, Serials, Structural Biology, Protein Biochemistry, Protocol, Protein expression and purification, Humans, skin and connective tissue diseases, Protein biochemistry, Structural biology, X-ray crystallography, Crystallization, Coronavirus 3C Proteases
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)., Graphical 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.

Published
2022

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