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 (1) , Julius Blechar , Anja Böckmann (2) , Louis Brigandat , Anna Bula , Matthias Bütikofer , Aldo R Camacho-Zarco (1) , 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 (1) , 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 (1) , Luiza Mamigonian Bessa (1) , Rachel W Martin , Tobias Matzel , Damien Maurin (1) , Seth W Mcnutt , Nathane Cunha Mebus-Antunes , Beat H Meier , Nathalie Meiser , Miguel Mompeán , Elisa Monaca , Roland Montserret , Laura Mariño Perez (1) , 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 (1) , 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 Á 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

1 IBS - UMR 5075 - Institut de biologie structurale
2 MMSB - Microbiologie moléculaire et biochimie structurale / Molecular Microbiology and Structural Biochemistry

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. Introduction

Keywords: COVID-19; SARS-CoV-2; nonstructural proteins; structural proteins; accessory proteins; intrinsically disordered region; cell-free protein synthesis; NMR spectroscopy

Domains: Virology; Structural Biology [q-bio.BM]; Genomics [q-bio.GN]; Biotechnology

Nadide Altincekic, Sophie Marianne Korn, Nusrat Shahin Qureshi, Marie Dujardin, Martí Ninot-Pedrosa, et al.. Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications. Frontiers in Molecular Biosciences, 2021, 8, pp.653148. ⟨10.3389/fmolb.2021.653148⟩. ⟨hal-03366994⟩