scholarly journals Rapid mass spectrometric peptide sequencing and mass matching for characterization of human melanoma proteins isolated by two-dimensional PAGE.

1995 ◽  
Vol 92 (11) ◽  
pp. 5072-5076 ◽  
Author(s):  
K. R. Clauser ◽  
S. C. Hall ◽  
D. M. Smith ◽  
J. W. Webb ◽  
L. E. Andrews ◽  
...  
Keyword(s):  
Peptide Sequencing ◽  
Human Melanoma ◽  
Mass Spectrometric ◽  
Two Dimensional ◽  
Rapid Mass

scholarly journals Mass spectrometric based detection of protein nucleotidylation in the RNA polymerase of SARS-CoV-2

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Brian J. Conti ◽  
Andrew S. Leicht ◽  
Robert N. Kirchdoerfer ◽  
Michael R. Sussman
Keyword(s):  
Rna Polymerase ◽  
Peptide Sequencing ◽  
Mass Spectrometric ◽  
Equine Arteritis Virus ◽  
Guanosine Monophosphate ◽  
Nucleotidyl Transferase ◽  
Replicase Proteins ◽  
New Strategies

AbstractCoronaviruses, like severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), encode a nucleotidyl transferase in the N-terminal (NiRAN) domain of the nonstructural protein (nsp) 12 protein within the RNA dependent RNA polymerase. Here we show the detection of guanosine monophosphate (GMP) and uridine monophosphate-modified amino acids in nidovirus proteins using heavy isotope-assisted mass spectrometry (MS) and MS/MS peptide sequencing. We identified lysine-143 in the equine arteritis virus (EAV) protein, nsp7, as a primary site of in vitro GMP attachment via a phosphoramide bond. In SARS-CoV-2 replicase proteins, we demonstrate nsp12-mediated nucleotidylation of nsp7 lysine-2. Our results demonstrate new strategies for detecting GMP-peptide linkages that can be adapted for higher throughput screening using mass spectrometric technologies. These data are expected to be important for a rapid and timely characterization of a new enzymatic activity in SARS-CoV-2 that may be an attractive drug target aimed at limiting viral replication in infected patients.

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