Enrichment and analysis of nonenzymatically glycated peptides: boronate affinity chromatography coupled with electron-transfer dissociation mass spectrometry.
Title | Enrichment and analysis of nonenzymatically glycated peptides: boronate affinity chromatography coupled with electron-transfer dissociation mass spectrometry. |
Publication Type | Journal Article |
Year of Publication | 2007 |
Authors | Zhang Q, Tang N, Brock JWC, Mottaz HM, Ames JM, Baynes JW, Smith RD, Metz TO |
Journal | J Proteome Res |
Keywords | Amino Acid Sequence, Boronic Acids, Chromatography, Affinity, Electron Transport, Glucose, Glycopeptides, Glycosylation, Mass Spectrometry, Molecular Sequence Data, Proteomics |
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 post-translational 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. |
DOI | 10.1021/pr070112q |
Alternate Journal | J. Proteome Res. |
PubMed ID | 17488106 |
PubMed Central ID | PMC2587408 |
Grant List | 06564 / / Wellcome Trust / United Kingdom 5R21DK071283 / DK / NIDDK NIH HHS / United States DK-19971 / DK / NIDDK NIH HHS / United States P41 RR018522-05 / RR / NCRR NIH HHS / United States R33 DK071283-03 / DK / NIDDK NIH HHS / United States RR018522 / RR / NCRR NIH HHS / United States |