Enrichment and analysis of nonenzymatically glycated peptides: Boronate affinity chromatography coupled with electron-transfer dissociation mass spectrometry

Q.B. Zhang, N. Tang, J.W.C. Brock, H.M. Mottaz, Jennifer Ames, John Baynes, R.D. Smith, T.O. Metz

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

Nonenzymatic glycation of peptides and proteins by D-glucose has important implications in the pathogenesis of diabetes mellitus, particularly in the development of diabetic complications. However, no effective high-throughput methods exist for identifying proteins containing this low-abundance posttranslational modification in bottom-up proteomic studies. In this report, phenylboronate affinity chromatography was used in a two-step enrichment scheme to selectively isolate first glycated proteins and then glycated, tryptic peptides from human serum glycated in vitro. Enriched peptides were subsequently analyzed by alternating electron-transfer dissociation (ETD) and collision induced dissociation ( CID) tandem mass spectrometry. ETD fragmentation mode permitted identification of a significantly higher number of glycated peptides (87.6% of all identified peptides) versus CID mode (17.0% of all identified peptides), when utilizing enrichment on first the protein and then the peptide level. This study illustrates that phenylboronate affinity chromatography coupled with LC-MS/MS and using ETD as the fragmentation mode is an efficient approach for analysis of glycated proteins and may have broad application in studies of diabetes mellitus.
Original languageEnglish
Pages (from-to)2323-2330
Number of pages8
JournalJournal of Proteome Research
Volume6(6)
Issue number6
DOIs
Publication statusPublished - Jun 2007

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics

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