Details
| Original language | English |
|---|---|
| Article number | 653148 |
| Journal | Frontiers in Molecular Biosciences |
| Volume | 8 |
| Publication status | Published - 10 May 2021 |
Abstract
The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.
Keywords
- accessory proteins, cell-free protein synthesis, COVID-19, intrinsically disordered region, NMR spectroscopy, nonstructural proteins, SARS-CoV-2, structural proteins
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry, Genetics and Molecular Biology (miscellaneous)
Sustainable Development Goals
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In: Frontiers in Molecular Biosciences, Vol. 8, 653148, 10.05.2021.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications
AU - Altincekic, Nadide
AU - Korn, Sophie Marianne
AU - Qureshi, Nusrat Shahin
AU - Dujardin, Marie
AU - Ninot-Pedrosa, Martí
AU - Abele, Rupert
AU - Abi Saad, Marie Jose
AU - Alfano, Caterina
AU - Almeida, Fabio C.L.
AU - Alshamleh, Islam
AU - de Amorim, Gisele Cardoso
AU - Anderson, Thomas K.
AU - Anobom, Cristiane D.
AU - Anorma, Chelsea
AU - Bains, Jasleen Kaur
AU - Bax, Adriaan
AU - Blackledge, Martin
AU - Blechar, Julius
AU - Böckmann, Anja
AU - Brigandat, Louis
AU - Bula, Anna
AU - Bütikofer, Matthias
AU - Camacho-Zarco, Aldo R.
AU - Carlomagno, Teresa
AU - Caruso, Icaro Putinhon
AU - Ceylan, Betül
AU - Chaikuad, Apirat
AU - Chu, Feixia
AU - Cole, Laura
AU - Crosby, Marquise G.
AU - de Jesus, Vanessa
AU - Dhamotharan, Karthikeyan
AU - Felli, Isabella C.
AU - Ferner, Jan
AU - Fleischmann, Yanick
AU - Fogeron, Marie Laure
AU - Fourkiotis, Nikolaos K.
AU - Fuks, Christin
AU - Fürtig, Boris
AU - Gallo, Angelo
AU - Gande, Santosh L.
AU - Gerez, Juan Atilio
AU - Ghosh, Dhiman
AU - Gomes-Neto, Francisco
AU - Gorbatyuk, Oksana
AU - Guseva, Serafima
AU - Hacker, Carolin
AU - Häfner, Sabine
AU - Hao, Bing
AU - Hargittay, Bruno
AU - Henzler-Wildman, K.
AU - Hoch, Jeffrey C.
AU - Hohmann, Katharina F.
AU - Hutchison, Marie T.
AU - Jaudzems, Kristaps
AU - Jović, Katarina
AU - Kaderli, Janina
AU - Kalniņš, Gints
AU - Kaņepe, Iveta
AU - Kirchdoerfer, Robert N.
AU - Kirkpatrick, John
AU - Knapp, Stefan
AU - Krishnathas, Robin
AU - Kutz, Felicitas
AU - zur Lage, Susanne
AU - Lambertz, Roderick
AU - Lang, Andras
AU - Laurents, Douglas
AU - Lecoq, Lauriane
AU - Linhard, Verena
AU - Löhr, Frank
AU - Malki, Anas
AU - Bessa, Luiza Mamigonian
AU - Martin, Rachel W.
AU - Matzel, Tobias
AU - Maurin, Damien
AU - McNutt, Seth W.
AU - Mebus-Antunes, Nathane Cunha
AU - Meier, Beat H.
AU - Meiser, Nathalie
AU - Mompeán, Miguel
AU - Monaca, Elisa
AU - Montserret, Roland
AU - Mariño Perez, Laura
AU - Moser, Celine
AU - Muhle-Goll, Claudia
AU - Neves-Martins, Thais Cristtina
AU - Ni, Xiamonin
AU - Norton-Baker, Brenna
AU - Pierattelli, Roberta
AU - Pontoriero, Letizia
AU - Pustovalova, Yulia
AU - Ohlenschläger, Oliver
AU - Orts, Julien
AU - Da Poian, Andrea T.
AU - Pyper, Dennis J.
AU - Richter, Christian
AU - Riek, Roland
AU - Rienstra, Chad M.
AU - Robertson, Angus
AU - Pinheiro, Anderson S.
AU - Sabbatella, Raffaele
AU - Salvi, Nicola
AU - Saxena, Krishna
AU - Schulte, Linda
AU - Schiavina, Marco
AU - Schwalbe, Harald
AU - Silber, Mara
AU - Almeida, Marcius da Silva
AU - Sprague-Piercy, Marc A.
AU - Spyroulias, Georgios A.
AU - Sreeramulu, Sridhar
AU - Tants, Jan Niklas
AU - Tārs, Kaspars
AU - Torres, Felix
AU - Töws, Sabrina
AU - Treviño, Miguel
AU - Trucks, Sven
AU - Tsika, Aikaterini C.
AU - Varga, Krisztina
AU - Wang, Ying
AU - Weber, Marco E.
AU - Weigand, Julia E.
AU - Wiedemann, Christoph
AU - Wirmer-Bartoschek, Julia
AU - Wirtz Martin, Maria Alexandra
AU - Zehnder, Johannes
AU - Hengesbach, Martin
AU - Schlundt, Andreas
N1 - Funding Information: The authors thank Leonardo Gonnelli and Katharina Targaczewski for the valuable technical assistance. IBS acknowledges integration into the Interdisciplinary Research Institute of Grenoble (IRIG CEA). They acknowledge the Advanced Technologies Network Center of the University of Palermo to support infrastructures. Funding Information: This work was supported by Goethe University (Corona funds), the DFG-funded CRC: “Molecular Principles of RNA-Based Regulation,” DFG infrastructure funds (project numbers: 277478796, 277479031, 392682309, 452632086, 70653611), the state of Hesse (BMRZ), the Fondazione CR Firenze (CERM), and the IWB-EFRE-program 20007375. This project has received funding from the European Union’s Horizon 2020 research and innovation program under Grant Agreement No. 871037. AS is supported by DFG Grant SCHL 2062/2-1 and by the JQYA at Goethe through project number 2019/AS01. Work in the lab of KV was supported by a CoRE grant from the University of New Hampshire. The FLI is a member of the Leibniz Association (WGL) and financially supported by the Federal Government of Germany and the State of Thuringia. Work in the lab of RM was supported by NIH (2R01EY021514) and NSF (DMR-2002837). BN-B was supported by the NSF GRFP. MC was supported by NIH (R25 GM055246 MBRS IMSD), and MS-P was supported by the HHMI Gilliam Fellowship. Work in the labs of KJ and KT was supported by Latvian Council of Science Grant No. VPP-COVID 2020/1-0014. Work in the UPAT’s lab was supported by the INSPIRED (MIS 5002550) project, which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure,” funded by the Operational Program “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014–2020) and cofinanced by Greece and the EU (European Regional Development Fund) and the FP7 REGPOT CT-2011-285950–“SEE-DRUG” project (purchase of UPAT’s 700 MHz NMR equipment). Work in the CM-G lab was supported by the Helmholtz society. Work in the lab of ABö was supported by the CNRS, the French National Research Agency (ANR, NMR-SCoV2-ORF8), the Fondation de la Recherche Médicale (FRM, NMR-SCoV2-ORF8), and the IR-RMN-THC Fr3050 CNRS. Work in the lab of BM was supported by the Swiss National Science Foundation (Grant number 200020_188711), the Günthard Stiftung für Physikalische Chemie, and the ETH Zurich. Work in the labs of ABö and BM was supported by a common grant from SNF (grant 31CA30_196256). This work was supported by the ETH Zurich, the grant ETH 40 18 1, and the grant Krebsliga KFS 4903 08 2019. Work in the lab of the IBS Grenoble was supported by the Agence Nationale de Recherche (France).
PY - 2021/5/10
Y1 - 2021/5/10
N2 - The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.
AB - The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium’s collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.
KW - accessory proteins
KW - cell-free protein synthesis
KW - COVID-19
KW - intrinsically disordered region
KW - NMR spectroscopy
KW - nonstructural proteins
KW - SARS-CoV-2
KW - structural proteins
UR - http://www.scopus.com/inward/record.url?scp=85105502063&partnerID=8YFLogxK
U2 - 10.3389/fmolb.2021.653148
DO - 10.3389/fmolb.2021.653148
M3 - Article
AN - SCOPUS:85105502063
VL - 8
JO - Frontiers in Molecular Biosciences
JF - Frontiers in Molecular Biosciences
M1 - 653148
ER -