Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications
Nadide Altincekic, Sophie Marianne Korn, Nusrat S. Qureshi, Marie Dujardin, Martí Ninot‐Pedrosa, Rupert Abele, Marie Jose Abi Saad, Caterina Alfano, Fábio C. L. Almeida, Islam Alshamleh, Gisele Cardoso de Amorim, Thomas K. Anderson, Cristiane Dinis Anobom, Chelsea Anorma, Jasleen Kaur Bains, Ad Bax, Martin Blackledge, Julius Blechar, Anja Böckmann, Louis Brigandat, Anna L. Bula, Matthias Bütikofer, Aldo R. Camacho‐Zarco, Teresa Carlomagno, Ícaro Putinhon Caruso, Betül Ceylan, A. 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, S.L. Gande, Juan Gerez, Dhiman Ghosh, Francisco Gomes‐Neto, Oksana Gorbatyuk, Serafima Guseva, Carolin Hacker, Sabine Häfner, Bing Hao, Bruno Hargittay, Katherine A. Henzler‐Wildman, Jeffrey C. Hoch, Katharina F. Hohmann, Marie Hutchison, Kristaps Jaudzems, Katarina Jović, Janina Kaderli, G. Kalnins, I. Kanepe, Robert N. Kirchdoerfer, John Kirkpatrick, Stefan Knapp, Robin Krishnathas, Felicitas Kutz, Susanne zur Lage, Roderick Lambertz, András Láng, Douglas V. Laurents, Lauriane Lecoq, V.L. Linhard, Frank Löhr, Anas Malki, Luiza M. Bessa, Rachel W. Martin, Tobias Matzel, Damien Maurin, Seth McNutt, Nathane C. Mebus-Antunes, Beat H. Meier, Nathalie Meiser, Miguel Mompeán, Elisa Monaca, Roland Montserret, Laura Mariño, Céline Moser, Claudia Muhle‐Goll, Thais C. 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 J. Robertson
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.