Backbone chemical shift assignments for the SARS-CoV-2 non-structural protein Nsp9: intermediate (ms – μs) dynamics in the C-terminal helix at the dimer interface
| Autor: | Mowei Zhou, Wesley C. Van Voorhis, Justin K. Craig, Garry W. Buchko, Peter J. Myler |
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| Rok vydání: | 2021 |
| Předmět: |
Magnetic Resonance Spectroscopy
Dimer 030303 biophysics Protein domain Protein dynamics Viral Nonstructural Proteins Crystallography X-Ray Ligands Biochemistry Protein Structure Secondary Article 03 medical and health sciences chemistry.chemical_compound Non-structural protein Native mass spectrometry Protein Domains Structural Biology Viral replication Disulfides Codon 030304 developmental biology Solution NMR Carbon Isotopes 0303 health sciences Binding Sites Nitrogen Isotopes SARS-CoV-2 Chemistry Chemical shift RNA-Binding Proteins Nuclear magnetic resonance spectroscopy Hydrogen-Ion Concentration Kinetics Crystallography Intramolecular force Helix Dimerization Heteronuclear single quantum coherence spectroscopy Hydrogen Protein Binding |
| Zdroj: | Biomolecular Nmr Assignments |
| ISSN: | 1874-270X 1874-2718 |
| DOI: | 10.1007/s12104-020-09992-1 |
| Popis: | The Betacoronavirus SARS-CoV-2 non-structural protein Nsp9 is a 113-residue protein that is essential for viral replication, and consequently, a potential target for the development of therapeutics against COVID19 infections. To capture insights into the dynamics of the protein’s backbone in solution and accelerate the identification and mapping of ligand-binding surfaces through chemical shift perturbation studies, the backbone 1H, 13C, and 15N NMR chemical shifts for Nsp9 have been extensively assigned. These assignments were assisted by the preparation of an ~ 70% deuterated sample and residue-specific, 15N-labelled samples (V, L, M, F, and K). A major feature of the assignments was the “missing” amide resonances for N96-L106 in the 1H-15N HSQC spectrum, a region that comprises almost the complete C-terminal α-helix that forms a major part of the homodimer interface in the crystal structure of SARS-CoV-2 Nsp9, suggesting this region either undergoes intermediate motion in the ms to μs timescale and/or is heterogenous. These “missing” amide resonances do not unambiguously appear in the 1H-15N HSQC spectrum of SARS-CoV-2 Nsp9 collected at a concentration of 0.0007 mM. At this concentration, at the detection limit, native mass spectrometry indicates the protein is exclusively in the monomeric state, suggesting the intermediate motion in the C-terminal of Nsp9 may be due to intramolecular dynamics. Perhaps this intermediate ms to μs timescale dynamics is the physical basis for a previously suggested “fluidity” of the C-terminal helix that may be responsible for homophilic (Nsp9-Nsp9) and postulated heterophilic (Nsp9-Unknown) protein-protein interactions. |
| Databáze: | OpenAIRE |
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