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

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

Research output: Contribution to journalArticle

26 Citations (Scopus)

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 1 2012

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Glycosylation
alpha-Macroglobulins
Polysaccharides
Glycopeptides
Electrons
Acetylglucosaminidase
Serum

All Science Journal Classification (ASJC) codes

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

Cite this

Lin, Zhenxin ; Lo, Andy ; Simeone, Diane M. ; Ruffin, Mack ; Lubman, David M. / An N-glycosylation analysis of human alpha-2-macroglobulin using an integrated approach. In: Journal of Proteomics and Bioinformatics. 2012 ; Vol. 5, No. 5. pp. 127-134.
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An N-glycosylation analysis of human alpha-2-macroglobulin using an integrated approach. / Lin, Zhenxin; Lo, Andy; Simeone, Diane M.; Ruffin, Mack; Lubman, David M.

In: Journal of Proteomics and Bioinformatics, Vol. 5, No. 5, 01.05.2012, p. 127-134.

Research output: Contribution to journalArticle

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