Combining protein identification and quantification: C-terminal isotope-coded tagging using sulfanilic acid.

Détails

ID Serval
serval:BIB_AB6E02BA9E5A
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Combining protein identification and quantification: C-terminal isotope-coded tagging using sulfanilic acid.
Périodique
Rapid Communications in Mass Spectrometry
Auteur(s)
Panchaud A., Guillaume E., Affolter M., Robert F., Moreillon P., Kussmann M.
ISSN
0951-4198[print], 0951-4198[linking]
Statut éditorial
Publié
Date de publication
2006
Volume
20
Numéro
10
Pages
1585-1594
Langue
anglais
Résumé
Two methods of differential isotopic coding of carboxylic groups have been developed to date. The first approach uses d0- or d3-methanol to convert carboxyl groups into the corresponding methyl esters. The second relies on the incorporation of two 18O atoms into the C-terminal carboxylic group during tryptic digestion of proteins in H(2)18O. However, both methods have limitations such as chromatographic separation of 1H and 2H derivatives or overlap of isotopic distributions of light and heavy forms due to small mass shifts. Here we present a new tagging approach based on the specific incorporation of sulfanilic acid into carboxylic groups. The reagent was synthesized in a heavy form (13C phenyl ring), showing no chromatographic shift and an optimal isotopic separation with a 6 Da mass shift. Moreover, sulfanilic acid allows for simplified fragmentation in matrix-assisted laser desorption/ionization (MALDI) due the charge fixation of the sulfonate group at the C-terminus of the peptide. The derivatization is simple, specific and minimizes the number of sample treatment steps that can strongly alter the sample composition. The quantification is reproducible within an order of magnitude and can be analyzed either by electrospray ionization (ESI) or MALDI. Finally, the method is able to specifically identify the C-terminal peptide of a protein by using GluC as the proteolytic enzyme.
Mots-clé
Caseins/chemistry, Computer Simulation, Data Interpretation, Statistical, Databases, Protein, Hydrolysis, Indicators and Reagents, Isotope Labeling, Opioid Peptides/chemistry, Peptides/chemistry, Peptides/isolation & purification, Proteins/analysis, Proteins/chemistry, Reproducibility of Results, Spectrometry, Mass, Electrospray Ionization, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Substance P/chemistry, Sulfanilic Acids/chemistry
Pubmed
Web of science
Création de la notice
24/01/2008 14:57
Dernière modification de la notice
20/08/2019 16:15
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