Author
Affilliation
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 SARSCoV-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 highthroughput drug screening. Here, we present the extensive catalog of a holistic SARSCoV-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.
Publisher
Frontiers Media
Citation
ALTINCEKIC, Nadide et al. Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications. Frontiers in Molecular Biosciences, v. 8, Article 653148, 25 p, May 2021.
DOI
10.3389/fmolb.2021.653148
ISSN
2296-889X
Notes
Francisco Gomes-Neto. Oswaldo Cruz Foundation. Oswaldo Cruz Instituto. Laboratory of Toxinology. Rio de Janeiro, RJ, Brazil. Institute for Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany, 2 Center of Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, Frankfurt am Main, Germany, 3 Institute for Molecular Biosciences, Goethe University Frankfurt, Frankfurt am Main, Germany, 4 Molecular Microbiology and Structural Biochemistry, UMR 5086, CNRS/Lyon University, Lyon, France, 5 Institute for Biochemistry, Goethe University Frankfurt, Frankfurt am Main, Germany, 6 Swiss Federal Institute of Technology, Laboratory of Physical Chemistry, ETH Zurich, Zurich, Switzerland, 7 Structural Biology and Biophysics Unit, Fondazione Ri.MED, Palermo, Italy, 8 National Center of Nuclear Magnetic Resonance (CNRMN, CENABIO), Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, 9 Institute of Medical Biochemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, 10 Multidisciplinary Center for Research in Biology (NUMPEX), Campus Duque de Caxias Federal University of Rio de Janeiro, Duque de Caxias, Brazil, 11 Institute for Molecular Virology, University of Wisconsin-Madison, Madison, WI, United States, 12 Institute of Chemistry, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil, 13 Department of Chemistry, University of California, Irvine, CA, United States, 14 LCP, NIDDK, NIH, Bethesda, MD, United States, 15 Univ. Grenoble Alpes, CNRS, CEA, IBS, Grenoble, France, 16 Latvian Institute of Organic Synthesis, Riga, Latvia, 17 BMWZ and Institute of Organic, Chemistry, Leibniz University Hannover, Hannover, Germany, 18 Group of NMR-Based Structural Chemistry, Helmholtz Centre for Infection Research, Braunschweig, Germany, 19 Multiuser Center for Biomolecular Innovation (CMIB), Department of Physics, São Paulo State University (UNESP), São José do Rio Preto, Brazil, 20 Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Frankfurt am Main, Germany, 21 Structural Genomics Consortium, Buchmann Institute for Molecular Life Sciences, Frankfurt am Main, Germany, 22 Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, NH, United States, 23 Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, United States, 24 Magnetic Resonance Centre (CERM), University of Florence, Sesto Fiorentino, Italy, 25 Department of Chemistry “Ugo Schiff”, University of Florence, Sesto Fiorentino, Italy, 26 Department of Pharmacy, University of Patras, Patras, Greece, 27 Laboratory of Toxinology, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil, 28 Department of Molecular Biology and Biophysics, UConn Health, Farmington, CT, United States, 29 Signals GmbH & Co. KG, Frankfurt am Main, Germany, 30 Leibniz Institute on Aging—Fritz Lipmann Institute (FLI), Jena, Germany, 31 Latvian Biomedical Research and Study Centre, Riga, Latvia, 32 “Rocasolano” Institute for Physical Chemistry (IQFR), Spanish National Research Council (CSIC), Madrid, Spain, 33 Institute of Biophysical Chemistry, Goethe University Frankfurt, Frankfurt am Main, Germany, 34 IBG-4, Karlsruhe Institute of Technology, Karlsruhe, Germany, 35 Department of Biochemistry and National Magnetic Resonance Facility at Madison, University of Wisconsin-Madison, Madison, WI, United States, 36 Department of Biology, Technical University of Darmstadt, Darmstadt, Germany, 37 Institute of Biochemistry and Biotechnology, Charles Tanford Protein Centre, Martin Luther University Halle-Wittenberg, Halle/Saale, Germany.
Share