Worldwide Network Develops SARS-CoV-2 Protocols for Research Laboratories

Scientists Martin Hengesbach (left) and Andreas Schlundt of the nuclear magnetic resonance (NMR) spectrometer at Goethe University Frankfurt, Germany. Credit: Uwe Dettmar for Goethe University Frankfurt, Germany

80% of all SARS-CoV-2 proteins. produced in the laboratory – protocols available for worldwide research – Goethe University Frankfurt, Germany, forms the center of research network from 17 countries.

When the SARS-CoV-2 virus mutates, it only initially means that there is a change in the genetic blueprint. For example, the mutation can cause an amino acid to be exchanged at a specific location in a viral protein. To quickly assess the effect of this change, a three-dimensional image of the viral protein is extremely useful. This is because it shows whether the switch to amino acid affects the function of the protein – or how it interacts with a potential drug or antibody.

Researchers from Goethe University Frankfurt and TU Darmstadt began to network internationally from the very beginning of the pandemic. Their goal: to describe the three-dimensional structures of SARS-CoV-2 molecules using nuclear magnetic resonance spectroscopy (NMR). In NMR spectroscopy, molecules are first labeled with special types of atoms (isotopes) and then exposed to a strong magnetic field. NMR can then be used to look in detail and with high throughput at how potentially active compounds bind to viral proteins. This is done in the Center for Biomolecular Magnetic Resonance (BMRZ) at Goethe University and other locations. However, the basic requirement is to produce large quantities of the proteins with high purity and stability, and with their correct folding, for the large number of tests.

The network, coordinated by Professor Harald Schwalbe of the Institute of Organic Chemistry and Chemical Biology at Goethe University, extends around the world. The development of laboratory protocols for the production of proteins is already the second milestone. In addition to proteins, the virus consists of RNA and the consortium made all important RNA fragments of SARS-CoV-2 accessible last year. With the expertise of 129 colleagues, it is now possible to produce and purify 23 of the almost 30 proteins of SARS-CoV-2 completely or as relevant fragments “in the test tube” and in large quantities.

For this, the genetic information of these proteins was processed into small ring-shaped pieces of DNA (plasmids). These plasmids were then introduced into bacteria for protein production. Some special proteins have also been produced in cell-free systems. Whether these proteins were still correctly folded after their isolation and enrichment was confirmed by NMR spectroscopy, among other things.

Dr. Martin Hengesbach of the Institute for Organic Chemistry and Chemical Biology at Goethe University explains: “We have isolated functional units of the SARS-CoV-2 proteins so that their structure, function and interactions can now be characterized by ourselves. and others. In addition, our large consortium provides working protocols that enable laboratories around the world to work quickly and reproducibly on SARS-CoV-2 proteins as well as future mutants. Disseminating this work from the outset was one of our top priorities. In addition to the protocols, we also make the plasmids freely available. “

Dr. Andreas Schlundt of the Institute for Molecular Biosciences at Goethe University says: “Our work accelerates the global search for active substances: Scientific laboratories equipped for this work do not have to spend several months setting up and optimizing systems. for the production and research of SARS-CoV-2 proteins, but can now start their research work within two weeks thanks to our elaborated protocols. Given the numerous mutations of SARS-CoV-2 to come, it is particularly important to have access to reliable, fast and proven methods of studying the virus in the laboratory. This will, for example, also facilitate research into the so-called helper proteins of SARS-CoV-2, which have remained under research, but which also play a role in the development of mutations. “

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Meanwhile, work in the NMR consortium continues: the researchers are currently working hard to find out whether viral proteins can bind to potential drugs.

The research work was funded by the German Research Foundation and the Goethe Coronavirus Fund. The high logistical effort and constant communication of research results was supported by Signals, a spin-off company from Goethe University.

Partners:

Brazil

National Center for Nuclear Magnetic Resonance (CNRMN, CENABIO), Federal University of Rio de Janeiro, Brazilian Institute of Medical Biochemistry, Federal University of Rio de Janeiro, Brazil Multidisciplinary Center for Research in Biology (NUMPEX), Campus Duque de Caxias, Federal University of Rio de Janeiro, Duque de Caxias, Brazil Institute of Chemistry, Federal University of Rio de Janeiro, Brazil Multiuser Center for Biomolecular Innovation (CMIB), Department of Physics, São Paulo State University (UNESP), São José do Rio Preto, Brazil Laboratory of Toxicology, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil

France

Molecular Microbiology and Structural Biochemistry (MMSB), UMR 5086, CNRS / Lyon University, France Université Grenoble Alpes, CNRS, CEA, IBS, Grenoble, France Germany Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Germany Center for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, Germany Institute for Molecular Biosciences, Goethe University Frankfurt, Germany Institute for Biochemistry, Goethe University Frankfurt, Germany Institute for Pharmaceutical Chemistry, Goethe University Frankfurt, Germany Institute for Biophysical Chemistry, Goethe University Frankfurt, Germany BMWZ and Institute of Organic Chemistry, Leibniz University Hannover, Germany Group of NMR-based Structural Chemistry, Helmholtz Center for Infection Research, Braunschweig, Germany Structural Genomics Consortium, Buchmann Institute for Molecular Life Sciences (BMLS), Germany Signals GmbH & Co. KG, Frankfurt am Main, Germany Leibniz Institute on Aging – Fritz Lipman n Institute (FLI), Jena, Germany IBG-4, Karlsruhe Institute of Technology, Karlsruhe, Germany Department of Biology, Technical University of Darmstadt, Darmstadt, Germany Institute of Biochemistry and Biotechnology, Charles Tanford Protein Center, Martin Luther University Halle-Wittenberg, Halle / Saale, Germany.

Greece

Department of Pharmacy, University of Patras, Greece

Italy

Structural Biology and Biophysics Unit, Fondazione Ri.MED, Palermo, Italy Magnetic Resonance Center (CERM), University of Florence, Sesto Fiorentino, Italy Department of Chemistry “Ugo Schiff”, University of Florence, Sesto Fiorentino, Italy

Latvia

Latvian Biomedical Research and Study Center, Riga, Latvia Latvian Institute of Organic Synthesis, Riga, Latvia

Switzerland

Swiss Federal Institute of Technology, Laboratory of Physical Chemistry, ETH Zurich, Zurich, Switzerland Spain “Rocasolano” Institute of Physical Chemistry (IQFR), Spanish National Research Council (CSIC), Serrano, Spain

USA

Institute for Molecular Virology, University of Wisconsin-Madison, WI, United States Department of Chemistry, University of California, Irvine, United States Laboratory of Chemical Physics, National Institute of Diabetes and Digestive Kidney Diseases, National Institute of Health, United States Department of Molecular, Cellular, and Biomedical Sciences, University of New Hampshire, Durham, NH, United States Department of Molecular Biology and Biochemistry, University of California, Irvine, California, United States Department of Molecular Biology and Biophysics, UC 72 onn Health, Farmington, CT , United States