Isotope-coded N-terminal sulfonation of peptides allows quantitative proteomic analysis with increased de novo peptide sequencing capability

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Recently various methods for the N-terminal sulfonation of peptides have been developed for the mass spectrometric analyses of proteomic samples to facilitate de novo sequencing of the peptides produced. This paper describes the isotope-coded N-terminal sulfonation (lCenS) of peptides; this procedure allows both de novo peptide sequencing and quantitative proteomics to be studied simultaneously. As N-terminal sulfonation reagents, C-13-labeled 4-sulfophenyl[(13)C6]isothiocyanate (C-13-SPITC) and unlabeled 4-sulfophenyl isothiocyanate (C-12-SPITC) were synthesized. The experimental and reference peptide mixtures were derivatized independently using C-13-SPITC and C-12-SPITC and then combined to generate an isotopically labeled peptide mixture in which each isotopic pair differs in mass by 6 Da. Capillary reverse-phase liquid chromatography/tandem mass spectrometry experiments on the resulting peptide mixtures revealed several immediate advantages of lCenS in addition to the de novo sequencing capability of N-terminal sulfonation, namely, differentiation between N-terminal sulfonated peptides and unmodified peptides in mass spectra, differentiation between N- and C-terminal fragments in tandem mass spectra of multiply protonated peptides by comparing fragmentations of the isotopic pairs, and relative peptide quantification between proteome samples. We demonstrate that the combination of N-terminal sulfonation and isotope coding in the mass spectrometric analysis of proteomic samples is a viable method that overcomes many problems associated with current N-terminal sulfonation methods. Copyright (C) 2004 John Wiley Sons, Ltd.
Publisher
JOHN WILEY SONS LTD
Issue Date
2004
Language
English
Article Type
Article
Keywords

FLIGHT MASS-SPECTROMETRY; EXPRESSION PROTEOMICS; LIQUID-CHROMATOGRAPHY; PROTEIN MIXTURES; AFFINITY TAGS; CELL-CULTURE; AMINO-ACIDS; DERIVATIZATION; SILAC; TOOL

Citation

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, v.18, no.24, pp.3019 - 3027

ISSN
0951-4198
DOI
10.1002/rcm.1724
URI
http://hdl.handle.net/10203/83657
Appears in Collection
CH-Journal Papers(저널논문)
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