Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications

Nadide Altincekic; Sophie Marianne Korn; Nusrat Shahin Qureshi; Marie Dujardin; Martí Ninot-Pedrosa; Rupert Abele; Marie Jose Abi Saad; Caterina Alfano; Fabio C.L. Almeida; Islam Alshamleh; Gisele Cardoso de Amorim; Thomas K. Anderson; Cristiane D. Anobom; Chelsea Anorma; Jasleen Kaur Bains; Adriaan Bax; Martin Blackledge; Julius Blechar; Anja Böckmann; Louis Brigandat; Anna Bula; Matthias Bütikofer; Aldo R. Camacho-Zarco; Teresa Carlomagno; Icaro Putinhon Caruso; Betül Ceylan; Apirat Chaikuad; Feixia Chu; Laura Cole; Marquise G. Crosby; Vanessa de Jesus; Karthikeyan Dhamotharan; Isabella C. Felli; Jan Ferner; Yanick Fleischmann; Marie Laure Fogeron; Nikolaos K. Fourkiotis; Christin Fuks; Boris Fürtig; Angelo Gallo; Santosh L. Gande; Juan Atilio Gerez; Dhiman Ghosh; Francisco Gomes-Neto; Oksana Gorbatyuk; Serafima Guseva; Carolin Hacker; Sabine Häfner; Bing Hao; Bruno Hargittay; K. Henzler-Wildman; Jeffrey C. Hoch; Katharina F. Hohmann; Marie T. Hutchison; Kristaps Jaudzems; Katarina Jović; Janina Kaderli; Gints Kalniņš; Iveta Kaņepe; Robert N. Kirchdoerfer; John Kirkpatrick; Stefan Knapp; Robin Krishnathas; Felicitas Kutz; Susanne zur Lage; Roderick Lambertz; Andras Lang; Douglas Laurents; Lauriane Lecoq; Verena Linhard; Frank Löhr; Anas Malki; Luiza Mamigonian Bessa; Rachel W. Martin; Tobias Matzel; Damien Maurin; Seth W. McNutt; Nathane Cunha Mebus-Antunes; Beat H. Meier; Nathalie Meiser; Miguel Mompeán; Elisa Monaca; Roland Montserret; Laura Mariño Perez; Celine Moser; Claudia Muhle-Goll; Thais Cristtina Neves-Martins; Xiamonin Ni; Brenna Norton-Baker; Roberta Pierattelli; Letizia Pontoriero; Yulia Pustovalova; Oliver Ohlenschläger; Julien Orts; Andrea T. Da Poian; Dennis J. Pyper; Christian Richter; Roland Riek; Chad M. Rienstra; Angus Robertson; Anderson S. Pinheiro; Raffaele Sabbatella; Nicola Salvi; Krishna Saxena; Linda Schulte; Marco Schiavina; Harald Schwalbe; Mara Silber; Marcius da Silva Almeida; Marc A. Sprague-Piercy; Georgios A. Spyroulias; Sridhar Sreeramulu; Jan Niklas Tants; Kaspars Tārs; Felix Torres; Sabrina Töws; Miguel Treviño; Sven Trucks; Aikaterini C. Tsika; Krisztina Varga; Ying Wang; Marco E. Weber; Julia E. Weigand; Christoph Wiedemann; Julia Wirmer-Bartoschek; Maria Alexandra Wirtz Martin; Johannes Zehnder; Martin Hengesbach; Andreas Schlundt

doi:10.3389/fmolb.2021.653148

Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications

Nadide Altincekic, Sophie Marianne Korn, Nusrat Shahin Qureshi, Marie Dujardin, Martí Ninot-Pedrosa, Rupert Abele, Marie Jose Abi Saad, Caterina Alfano, Fabio C.L. Almeida, Islam Alshamleh, Gisele Cardoso de Amorim, Thomas K. Anderson, Cristiane D. Anobom, Chelsea Anorma, Jasleen Kaur Bains, Adriaan Bax, Martin Blackledge, Julius Blechar, Anja Böckmann^*, Louis BrigandatAnna Bula, Matthias Bütikofer, Aldo R. Camacho-Zarco, Teresa Carlomagno, Icaro Putinhon Caruso, Betül Ceylan, Apirat Chaikuad, Feixia Chu, Laura Cole, Marquise G. Crosby, Vanessa de Jesus, Karthikeyan Dhamotharan, Isabella C. Felli, Jan Ferner, Yanick Fleischmann, Marie Laure Fogeron, Nikolaos K. Fourkiotis, Christin Fuks, Boris Fürtig, Angelo Gallo, Santosh L. Gande, Juan Atilio Gerez, Dhiman Ghosh, Francisco Gomes-Neto, Oksana Gorbatyuk, Serafima Guseva, Carolin Hacker, Sabine Häfner, Bing Hao, Bruno Hargittay, K. Henzler-Wildman, Jeffrey C. Hoch, Katharina F. Hohmann, Marie T. Hutchison, Kristaps Jaudzems, Katarina Jović, Janina Kaderli, Gints Kalniņš, Iveta Kaņepe, Robert N. Kirchdoerfer, John Kirkpatrick, Stefan Knapp, Robin Krishnathas, Felicitas Kutz, Susanne zur Lage, Roderick Lambertz, Andras Lang, Douglas Laurents, Lauriane Lecoq, Verena Linhard, Frank Löhr, Anas Malki, Luiza Mamigonian Bessa, Rachel W. Martin, Tobias Matzel, Damien Maurin, Seth W. McNutt, Nathane Cunha Mebus-Antunes, Beat H. Meier, Nathalie Meiser, Miguel Mompeán, Elisa Monaca, Roland Montserret, Laura Mariño Perez, Celine Moser, Claudia Muhle-Goll, Thais Cristtina Neves-Martins, Xiamonin Ni, Brenna Norton-Baker, Roberta Pierattelli, Letizia Pontoriero, Yulia Pustovalova, Oliver Ohlenschläger, Julien Orts, Andrea T. Da Poian, Dennis J. Pyper, Christian Richter, Roland Riek, Chad M. Rienstra, Angus Robertson, Anderson S. Pinheiro, Raffaele Sabbatella, Nicola Salvi, Krishna Saxena, Linda Schulte, Marco Schiavina, Harald Schwalbe^*, Mara Silber, Marcius da Silva Almeida, Marc A. Sprague-Piercy, Georgios A. Spyroulias, Sridhar Sreeramulu, Jan Niklas Tants, Kaspars Tārs, Felix Torres, Sabrina Töws, Miguel Treviño, Sven Trucks, Aikaterini C. Tsika, Krisztina Varga, Ying Wang, Marco E. Weber, Julia E. Weigand, Christoph Wiedemann, Julia Wirmer-Bartoschek, Maria Alexandra Wirtz Martin, Johannes Zehnder, Martin Hengesbach^*, Andreas Schlundt

^*Corresponding author for this work

Biosciences

Research output: Contribution to journal › Article › peer-review

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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.

Original language	English
Article number	653148
Number of pages	25
Journal	Frontiers in Molecular Biosciences
Volume	8
DOIs	https://doi.org/10.3389/fmolb.2021.653148
Publication status	Published - 10 May 2021

Bibliographical note

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.

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)

Publisher Copyright:
© Copyright © 2021 Altincekic.

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
Molecular Biology
Biochemistry, Genetics and Molecular Biology (miscellaneous)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

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Cite this

Altincekic, N., Korn, S. M., Qureshi, N. S., Dujardin, M., Ninot-Pedrosa, M., Abele, R., Abi Saad, M. J., Alfano, C., Almeida, F. C. L., Alshamleh, I., de Amorim, G. C., Anderson, T. K., Anobom, C. D., Anorma, C., Bains, J. K., Bax, A., Blackledge, M., Blechar, J., Böckmann, A., ... Schlundt, A. (2021). Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications. Frontiers in Molecular Biosciences, 8, Article 653148. https://doi.org/10.3389/fmolb.2021.653148

@article{d02f95cfa8c4454b82b1110df37b7dc0,

title = "Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications",

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{\textquoteright}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",

author = "Nadide Altincekic and Korn, {Sophie Marianne} and Qureshi, {Nusrat Shahin} and Marie Dujardin and Mart{\'i} Ninot-Pedrosa and Rupert Abele and {Abi Saad}, {Marie Jose} and Caterina Alfano and Almeida, {Fabio C.L.} and Islam Alshamleh and {de Amorim}, {Gisele Cardoso} and Anderson, {Thomas K.} and Anobom, {Cristiane D.} and Chelsea Anorma and Bains, {Jasleen Kaur} and Adriaan Bax and Martin Blackledge and Julius Blechar and Anja B{\"o}ckmann and Louis Brigandat and Anna Bula and Matthias B{\"u}tikofer and Camacho-Zarco, {Aldo R.} and Teresa Carlomagno and Caruso, {Icaro Putinhon} and Bet{\"u}l Ceylan and Apirat Chaikuad and Feixia Chu and Laura Cole and Crosby, {Marquise G.} and {de Jesus}, Vanessa and Karthikeyan Dhamotharan and Felli, {Isabella C.} and Jan Ferner and Yanick Fleischmann and Fogeron, {Marie Laure} and Fourkiotis, {Nikolaos K.} and Christin Fuks and Boris F{\"u}rtig and Angelo Gallo and Gande, {Santosh L.} and Gerez, {Juan Atilio} and Dhiman Ghosh and Francisco Gomes-Neto and Oksana Gorbatyuk and Serafima Guseva and Carolin Hacker and Sabine H{\"a}fner and Bing Hao and Bruno Hargittay and K. Henzler-Wildman and Hoch, {Jeffrey C.} and Hohmann, {Katharina F.} and Hutchison, {Marie T.} and Kristaps Jaudzems and Katarina Jovi{\'c} and Janina Kaderli and Gints Kalniņ{\v s} and Iveta Kaņepe and Kirchdoerfer, {Robert N.} and John Kirkpatrick and Stefan Knapp and Robin Krishnathas and Felicitas Kutz and {zur Lage}, Susanne and Roderick Lambertz and Andras Lang and Douglas Laurents and Lauriane Lecoq and Verena Linhard and Frank L{\"o}hr and Anas Malki and Bessa, {Luiza Mamigonian} and Martin, {Rachel W.} and Tobias Matzel and Damien Maurin and McNutt, {Seth W.} and Mebus-Antunes, {Nathane Cunha} and Meier, {Beat H.} and Nathalie Meiser and Miguel Mompe{\'a}n and Elisa Monaca and Roland Montserret and {Mari{\~n}o Perez}, Laura and Celine Moser and Claudia Muhle-Goll and Neves-Martins, {Thais Cristtina} and Xiamonin Ni and Brenna Norton-Baker and Roberta Pierattelli and Letizia Pontoriero and Yulia Pustovalova and Oliver Ohlenschl{\"a}ger and Julien Orts and {Da Poian}, {Andrea T.} and Pyper, {Dennis J.} and Christian Richter and Roland Riek and Rienstra, {Chad M.} and Angus Robertson and Pinheiro, {Anderson S.} and Raffaele Sabbatella and Nicola Salvi and Krishna Saxena and Linda Schulte and Marco Schiavina and Harald Schwalbe and Mara Silber and Almeida, {Marcius da Silva} and Sprague-Piercy, {Marc A.} and Spyroulias, {Georgios A.} and Sridhar Sreeramulu and Tants, {Jan Niklas} and Kaspars Tārs and Felix Torres and Sabrina T{\"o}ws and Miguel Trevi{\~n}o and Sven Trucks and Tsika, {Aikaterini C.} and Krisztina Varga and Ying Wang and Weber, {Marco E.} and Weigand, {Julia E.} and Christoph Wiedemann and Julia Wirmer-Bartoschek and {Wirtz Martin}, {Maria Alexandra} and Johannes Zehnder and Martin Hengesbach and Andreas Schlundt",

note = "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. 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{\textquoteright}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{\textquoteright}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{\textquoteright}s 700 MHz NMR equipment). Work in the CM-G lab was supported by the Helmholtz society. Work in the lab of AB{\"o} was supported by the CNRS, the French National Research Agency (ANR, NMR-SCoV2-ORF8), the Fondation de la Recherche M{\'e}dicale (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 G{\"u}nthard Stiftung f{\"u}r Physikalische Chemie, and the ETH Zurich. Work in the labs of AB{\"o} 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) Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2021 Altincekic. ",

year = "2021",

month = may,

day = "10",

doi = "10.3389/fmolb.2021.653148",

language = "English",

volume = "8",

journal = "Frontiers in Molecular Biosciences",

issn = "2296-889X",

publisher = "Frontiers Media",

}

Altincekic, N, Korn, SM, Qureshi, NS, Dujardin, M, Ninot-Pedrosa, M, Abele, R, Abi Saad, MJ, Alfano, C, Almeida, FCL, Alshamleh, I, de Amorim, GC, Anderson, TK, Anobom, CD, Anorma, C, Bains, JK, Bax, A, Blackledge, M, Blechar, J, Böckmann, A, Brigandat, L, Bula, A, Bütikofer, M, Camacho-Zarco, AR, Carlomagno, T, Caruso, IP, Ceylan, B, Chaikuad, A, Chu, F, Cole, L, Crosby, MG, de Jesus, V, Dhamotharan, K, Felli, IC, Ferner, J, Fleischmann, Y, Fogeron, ML, Fourkiotis, NK, Fuks, C, Fürtig, B, Gallo, A, Gande, SL, Gerez, JA, Ghosh, D, Gomes-Neto, F, Gorbatyuk, O, Guseva, S, Hacker, C, Häfner, S, Hao, B, Hargittay, B, Henzler-Wildman, K, Hoch, JC, Hohmann, KF, Hutchison, MT, Jaudzems, K, Jović, K, Kaderli, J, Kalniņš, G, Kaņepe, I, Kirchdoerfer, RN, Kirkpatrick, J, Knapp, S, Krishnathas, R, Kutz, F, zur Lage, S, Lambertz, R, Lang, A, Laurents, D, Lecoq, L, Linhard, V, Löhr, F, Malki, A, Bessa, LM, Martin, RW, Matzel, T, Maurin, D, McNutt, SW, Mebus-Antunes, NC, Meier, BH, Meiser, N, Mompeán, M, Monaca, E, Montserret, R, Mariño Perez, L, Moser, C, Muhle-Goll, C, Neves-Martins, TC, Ni, X, Norton-Baker, B, Pierattelli, R, Pontoriero, L, Pustovalova, Y, Ohlenschläger, O, Orts, J, Da Poian, AT, Pyper, DJ, Richter, C, Riek, R, Rienstra, CM, Robertson, A, Pinheiro, AS, Sabbatella, R, Salvi, N, Saxena, K, Schulte, L, Schiavina, M, Schwalbe, H, Silber, M, Almeida, MDS, Sprague-Piercy, MA, Spyroulias, GA, Sreeramulu, S, Tants, JN, Tārs, K, Torres, F, Töws, S, Treviño, M, Trucks, S, Tsika, AC, Varga, K, Wang, Y, Weber, ME, Weigand, JE, Wiedemann, C, Wirmer-Bartoschek, J, Wirtz Martin, MA, Zehnder, J, Hengesbach, M & Schlundt, A 2021, 'Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications', Frontiers in Molecular Biosciences, vol. 8, 653148. https://doi.org/10.3389/fmolb.2021.653148

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. 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) Publisher Copyright: © Copyright © 2021 Altincekic.

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

C2 - 34041264

SN - 2296-889X

VL - 8

JO - Frontiers in Molecular Biosciences

JF - Frontiers in Molecular Biosciences

M1 - 653148

ER -

Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications

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