Refine
Year of publication
Language
- English (22)
Has Fulltext
- yes (22)
Is part of the Bibliography
- no (22)
Keywords
- SARS-CoV-2 (2)
- nuclear magnetic resonance spectroscopy (2)
- ABC transporter (1)
- ABC transporters (1)
- Biophysical chemistry (1)
- COVID-19 (1)
- Covid19-NMR (1)
- Dimethyl maleic anhydride (1)
- Enzyme mechanisms (1)
- LILBID-MS (1)
- Membrane protein complex (1)
- Membrane proteins (1)
- Methanogenesis (1)
- Mycobacterium tuberculosis (1)
- N5 -methyl-tetrahydromethanopterin: coenzyme M methyltransferase (1)
- NMR spectroscopy (1)
- Non-structural protein (1)
- Protein folding (1)
- RNA processing (1)
- RRM domain (1)
- SAP domain (1)
- SEC-MALS (1)
- Solution NMR-spectroscopy (1)
- accessory proteins (1)
- antigen presentation (1)
- antigen processing (1)
- cell-free protein synthesis (1)
- cholesterol (1)
- chromatin (1)
- drug resistance (1)
- dual nucleic acid binding (1)
- endolysosomal system (1)
- endoplasmic reticulum (1)
- intrinsically disordered protein (1)
- intrinsically disordered region (1)
- lipid (1)
- lysosome (1)
- lysosomes (1)
- membrane proteins (1)
- nonstructural proteins (1)
- nuclear matrix (1)
- peptide transport (1)
- protein denaturation (1)
- protein dynamics (1)
- protein phosphorylation (1)
- protein tyrosine kinase (1)
- scaffold attachment factor proteins (1)
- structural proteins (1)
- surfactant (1)
- trafficking (1)
- viral immune escape (1)
- vitamin B12 (1)
1H, 13C, and 15N backbone chemical shift assignments of coronavirus-2 non-structural protein Nsp10
(2020)
The international Covid19-NMR consortium aims at the comprehensive spectroscopic characterization of SARS-CoV-2 RNA elements and proteins and will provide NMR chemical shift assignments of the molecular components of this virus. The SARS-CoV-2 genome encodes approximately 30 different proteins. Four of these proteins are involved in forming the viral envelope or in the packaging of the RNA genome and are therefore called structural proteins. The other proteins fulfill a variety of functions during the viral life cycle and comprise the so-called non-structural proteins (nsps). Here, we report the near-complete NMR resonance assignment for the backbone chemical shifts of the non-structural protein 10 (nsp10). Nsp10 is part of the viral replication-transcription complex (RTC). It aids in synthesizing and modifying the genomic and subgenomic RNAs. Via its interaction with nsp14, it ensures transcriptional fidelity of the RNA-dependent RNA polymerase, and through its stimulation of the methyltransferase activity of nsp16, it aids in synthesizing the RNA cap structures which protect the viral RNAs from being recognized by the innate immune system. Both of these functions can be potentially targeted by drugs. Our data will aid in performing additional NMR-based characterizations, and provide a basis for the identification of possible small molecule ligands interfering with nsp10 exerting its essential role in viral replication.
Methanogenic archaea share one ion gradient forming reaction in their energy metabolism catalyzed by the membrane-spanning multisubunit complex N5-methyl-tetrahydromethanopterin: coenzyme M methyltransferase (MtrABCDEFGH or simply Mtr). In this reaction the methyl group transfer from methyl-tetrahydromethanopterin to coenzyme M mediated by cobalamin is coupled with the vectorial translocation of Na+ across the cytoplasmic membrane. No detailed structural and mechanistic data are reported about this process. In the present work we describe a procedure to provide a highly pure and homogenous Mtr complex on the basis of a selective removal of the only soluble subunit MtrH with the membrane perturbing agent dimethyl maleic anhydride and a subsequent two-step chromatographic purification. A molecular mass determination of the Mtr complex by laser induced liquid bead ion desorption mass spectrometry (LILBID-MS) and size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS) resulted in a (MtrABCDEFG)3 heterotrimeric complex of ca. 430 kDa with both techniques. Taking into account that the membrane protein complex contains various firmly bound small molecules, predominantly detergent molecules, the stoichiometry of the subunits is most likely 1:1. A schematic model for the subunit arrangement within the MtrABCDEFG protomer was deduced from the mass of Mtr subcomplexes obtained by harsh IR-laser LILBID-MS.