Doublet N-Terminal Oriented Proteomics for N-Terminomics and Proteolytic Processing Identification.
The study of the N-terminome and the precise identification of proteolytic processing events are key in biology. Dedicated methodologies have been developed as the comprehensive characterization of the N-terminome can hardly be achieved by standard proteomics methods. In this context, we have set up a trimethoxyphenyl phosphonium (TMPP) labeling approach that allows the characterization of both N-terminal and internal digestion peptides in a single experiment. This latter point is a major advantage of our strategy as most N-terminomics methods rely on the enrichment of N-terminal peptides and thus exclude internal peptides.We have implemented a double heavy/light TMPP labeling and an automated data validation workflow that make our doublet N-terminal oriented proteomics (dN-TOP) strategy efficient for high-throughput N-terminome analysis.
BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, CNRS-UdS, UMR 7178, University of Strasbourg, 25, rue Becquerel, 67087, Strasbourg, France.
Benoit
B
Westermann
BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, CNRS-UdS, UMR 7178, University of Strasbourg, 25, rue Becquerel, 67087, Strasbourg, France.
Alvaro Sebastian Vaca
ASV
Jacome
BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, CNRS-UdS, UMR 7178, University of Strasbourg, 25, rue Becquerel, 67087, Strasbourg, France.
Magali
M
Rompais
BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, CNRS-UdS, UMR 7178, University of Strasbourg, 25, rue Becquerel, 67087, Strasbourg, France.
Christine
C
Carapito
BioOrganic Mass Spectrometry Laboratory (LSMBO), IPHC, CNRS-UdS, UMR 7178, University of Strasbourg, 25, rue Becquerel, 67087, Strasbourg, France. christine.schaeffer@unistra.fr.
Christine
C
Schaeffer-Reiss
D002853:Chromatography, Liquid / Q000379:methods*; D007553:Isotope Labeling; D010446:Peptide Fragments; D059748:Proteolysis; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D013223:Statistics as Topic / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D057188:Workflow
D010446:Peptide Fragments; D020543:Proteome
N-terminomics; Proteogenomics; TMPP derivatization; dN-TOP approach