Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography
Summary: 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...
| Main Authors: | , , , , , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722000387 |
| _version_ | 1830230565572313088 |
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| 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 |
| author_facet | 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 |
| author_sort | Fatma Betul Ertem |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-12-18T10:57:04Z |
| format | Article |
| id | doaj.art-57d9c8025fa44a7b88ba24db6f0d7586 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-18T10:57:04Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-57d9c8025fa44a7b88ba24db6f0d75862022-12-21T21:10:19ZengElsevierSTAR Protocols2666-16672022-03-0131101158Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallographyFatma Betul Ertem0Omur Guven1Cengizhan Buyukdag2Oktay Gocenler3Esra Ayan4Busra Yuksel5Mehmet Gul6Gozde Usta7Barıs Cakılkaya8J. Austin Johnson9E. Han Dao10Zhen Su11Frederic Poitevin12Chun Hong Yoon13Christopher Kupitz14Brandon Hayes15Mengning Liang16Mark S. Hunter17Alexander Batyuk18Raymond G. Sierra19Gihan Ketawala20Sabine Botha21Çağdaş Dağ22Hasan DeMirci23Department of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyDepartment of Molecular Biology and Genetics, Koc University, Istanbul, TurkeyLinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USA; Department of Applied Physics, Stanford University, Stanford, CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USALinac Coherent Light Source, SLAC National Accelerator Laboratory, 2575 Sand Hill Road, Menlo Park, 94025 CA, USADepartment of Physics, Arizona State University, Tempe, AZ 85287-1504, USA; Biodesign Center for Applied Structural Discovery, Arizona State University, Tempe, AZ 85287-5001, USADepartment of Physics, Arizona State University, Tempe, AZ 85287-1504, USA; Biodesign Center for Applied Structural Discovery, Arizona State University, Tempe, AZ 85287-5001, USADepartment of Molecular Biology and Genetics, Koc University, Istanbul, Turkey; Koc University, Nanofabrication and Nanocharacterization Center for Scientific and Technological Advanced Research (n2STAR), Istanbul, Turkey; Koc University Isbank Center for Infectious Diseases (KUISCID), Istanbul, TurkeyDepartment 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; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166722000387Protein BiochemistryProtein expression and purificationStructural BiologyX-ray Crystallography |
| spellingShingle | 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 Protocol for structure determination of SARS-CoV-2 main protease at near-physiological-temperature by serial femtosecond crystallography STAR Protocols Protein Biochemistry Protein expression and purification Structural Biology X-ray 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 | Protein Biochemistry Protein expression and purification Structural Biology X-ray Crystallography |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722000387 |
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