An N-glycosylation analysis of human alpha-2-macroglobulin using an integrated approach

Zhenxin Lin, Andy Lo, Diane M. Simeone, Mack T. Ruffin, David M. Lubman

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

Assignment of glycosylation sites and site microheterogeneity is of both biological and clinical significance. Herein, the detailed N-glycosylation pattern of human serum alpha-2-macroglobulin was studied using an integrative approach, including permethylation of N-glycans, collision induced dissociation (CID) and electron transfer dissociation (ETD) of chymotryptic N-glycopeptides, and partial deglycosylation of chymotryptic N-glycopeptides with endo-β-N-acetylglucosaminidase F3 (Endo F3). Three N-glycosylation sites were found to be occupied by four biantennary complex type N-glycans using N-glycan analysis and the ETD/CID method. Endo F3 assisted mass spectrometric analysis yielded five N-glycosylation sites with and without core fucosylation. In total, six out of eight potential N-glycosylation sites were identified using this approach. This integrative approach was performed using only 10 μL of human serum for both N-glycosylation site assignment and site microheterogeneity determination.

Original languageEnglish (US)
Pages (from-to)127-134
Number of pages8
JournalJournal of Proteomics and Bioinformatics
Volume5
Issue number5
DOIs
StatePublished - May 2012

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Computer Science Applications
  • Cell Biology

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