Differentially isotope-coded N-terminal protein sulphonation: combining protein identification and quantification

Proteomics
Elisabeth GuillaumeMartin Kussmann

Abstract

Most proteomic labelling technologies intend to improve protein quantification and/or facilitate (de novo) peptide sequencing. We present here a novel stable-isotope labelling method to simultaneously identify and quantify protein components in complex mixtures by specifically derivatizing the N-terminus of proteins with 4-sulphophenyl isothiocyanate (SPITC). Our approach combines protein identification with quantification through differential isotope-coded labelling at the protein N-terminus prior to digestion. The isotope spacing of 6 Da (unlabelled vs. six-fold 13C-labelled tag) between derivatized peptide pairs enables the detection on different MS platforms (MALDI and ESI). Optimisation of the reaction conditions using SPITC was performed on three model proteins. Improved detection of the N-terminally derivatized peptide compared to the native analogue was observed in negative-ion MALDI-MS. Simpler fragmentation patterns compared to native peptides facilitated protein identification. The 13C-labelled SPITC resulted in convenient peptide pair spacing without isotopic overlap and hence facilitated relative quantification by MALDI-TOF/TOF and LC-ESI-MS/MS. The combination of facilitated identification and quantification achie...Continue Reading

References

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Citations

Dec 28, 2011·Analytical and Bioanalytical Chemistry·Jong-Seo KimHie-Joon Kim
Nov 26, 2008·Journal of the American Society for Mass Spectrometry·Lisa A VasicekJennifer S Brodbelt
Jul 26, 2011·The Proceedings of the Nutrition Society·Sandra Sénéchal, Martin Kussmann
Aug 17, 2006·Current Opinion in Clinical Nutrition and Metabolic Care·Martin Kussmann, Michael Affolter
Mar 24, 2007·Clinical Chemistry and Laboratory Medicine : CCLM·Martin Kussmann
Nov 16, 2010·Journal of Proteomics·María Jesús García-MurriaManuel M Sánchez del Pino
Jun 11, 2008·Journal of Proteomics·Alexandre PanchaudMartin Kussmann
Aug 31, 2006·Rapid Communications in Mass Spectrometry : RCM·Chunxi ZhouXiaohong Qian
May 9, 2014·Journal of Mass Spectrometry : JMS·Wenlong ZhangSongping Liang
Apr 8, 2006·Journal of Biotechnology·Martin KussmannMichael Affolter
Dec 18, 2007·Molecular & Cellular Proteomics : MCP·Alexandre PanchaudMartin Kussmann
Oct 13, 2009·Journal of Proteome Research·Hong WangSamir Hanash

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