1 H, 13 C, and 15 N backbone chemical shift assignments of the apo and the ADP-ribose bound forms of the macrodomain of
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ARTICLE
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H, 13C, and 15N backbone chemical shift assignments of the apo and the ADP‑ribose bound forms of the macrodomain of SARS‑CoV‑2 non‑structural protein 3b F. Cantini1,2 · L. Banci1,2 · N. Altincekic3 · J. K. Bains3 · K. Dhamotharan4 · C. Fuks3 · B. Fürtig3 · S. L. Gande · B. Hargittay3 · M. Hengesbach3 · M. T. Hutchison3 · S. M. Korn4 · N. Kubatova3 · F. Kutz3 · V. Linhard3 · F. Löhr5 · N. Meiser3 · D. J. Pyper3 · N. S. Qureshi3 · C. Richter3 · K. Saxena3 · A. Schlundt4 · H. Schwalbe3 · S. Sreeramulu3 · J.‑N. Tants4 · A. Wacker3 · J. E. Weigand6 · J. Wöhnert4 · A. C. Tsika7 · N. K. Fourkiotis7 · G. A. Spyroulias7 Received: 13 July 2020 / Accepted: 31 July 2020 © The Author(s) 2020
Abstract The SARS-CoV-2 genome encodes for approximately 30 proteins. Within the international project COVID19-NMR, we distribute the spectroscopic analysis of the viral proteins and RNA. Here, we report NMR chemical shift assignments for the protein Nsp3b, a domain of Nsp3. The 217-kDa large Nsp3 protein contains multiple structurally independent, yet functionally related domains including the viral papain-like protease and Nsp3b, a macrodomain (MD). In general, the MDs of SARS-CoV and MERS-CoV were suggested to play a key role in viral replication by modulating the immune response of the host. The MDs are structurally conserved. They most likely remove ADP-ribose, a common posttranslational modification, from protein side chains. This de-ADP ribosylating function has potentially evolved to protect the virus from the anti-viral ADP-ribosylation catalyzed by poly-ADP-ribose polymerases (PARPs), which in turn are triggered by pathogen-associated sensing of the host immune system. This renders the SARS-CoV-2 Nsp3b a highly relevant drug target in the viral replication process. We here report the near-complete NMR backbone resonance assignment (1H, 13C, 15N) of the putative Nsp3b MD in its apo form and in complex with ADP-ribose. Furthermore, we derive the secondary structure of Nsp3b in solution. In addition, 15N-relaxation data suggest an ordered, rigid core of the MD structure. These data will provide a basis for NMR investigations targeted at obtaining small-molecule inhibitors interfering with the catalytic activity of Nsp3b. Keywords SARS-CoV-2 · Non-structural protein · Macrodomain · Solution NMR-spectroscopy · Protein drugability · COVID19-NMR
F. Cantini, K. Saxena and A. C. Tsika are joint first authors. * L. Banci [email protected]; covid19‑[email protected]‑frankfurt.de
Johann Wolfgang Goethe-University Frankfurt, Max‑von‑Laue‑Str. 7, 60438 Frankfurt, Germany 4
Institute for Molecular Biosciences, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-University Frankfurt, Max‑von‑Laue‑Str. 7, 60438 Frankfurt, Germany
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Magnetic Resonance Center – CERM, University of Florence, Via Luigi Sacconi 6, Sesto Fiorentino, 50019 Florence, Italy
Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-University Frankfurt, Max‑von‑Laue‑St
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