A simple carbamidomethylation-based isotope labeling method for quantitative shotgun proteomics

Donggeun Oh, Sun Young Lee, Meehyang Kwon, Sook Kyung Kim, Myeong Hee Moon, Dukjin Kang

Research output: Contribution to journalArticle

Abstract

In this study, we present a new isotope-coded carbamidomethylation (iCCM)-based quantitative proteomics, as a complementary strategy for conventional isotope labeling strategies, with providing the simplicity, ease of use, and robustness. In iCCM-based quantification, two proteome samples can be separately isotope-labeled by means of covalently reaction of all cysteinyl residues in proteins with iodoacetamide (IAA) and its isotope (IAA-13C2, D2), denoted as CM and iCCM, respectively, leading to a mass shift of all cysteinyl residues to be + 4 Da. To evaluate iCCM-based isotope labeling in proteomic quantification, 6 protein standards (i.e., bovine serum albumin, serotransferrin, lysozyme, beta-lactoglobulin, beta-galactosidase, and alpha-lactalbumin) isotopically labeled with IAA and its isotope, mixed equally, and followed by proteolytic digestion. The resulting CM-/iCCM-labeled peptide mixtures were analyzed using a nLC-ESI-FT orbitrap-MS/MS. From our experimental results, we found that the efficiency of iCCM-based quantification is more superior to that of mTRAQ, as a conventional nonisobaric labeling method, in which both of a number of identified peptides from 6 protein standards and the less quantitative variations in the relative abundance ratios of heavy-/light-labeled corresponding peptide pairs. Finally, we applied the developed iCCM-based quantitative method to lung cancer serum proteome in order to evaluate the potential in biomarker discovery study.

Original languageEnglish
Pages (from-to)63-69
Number of pages7
JournalMass Spectrometry Letters
Volume5
Issue number3
DOIs
Publication statusPublished - 2014 Sep 1

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Spectroscopy

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