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One-pipeline approach achieving glycoprotein identification and obtaining intact glycopeptide information by tandem mass spectrometry

Reputable Mentor II
Reputable Mentor II
Chen Y, Liu M, Yan G, Lu H, Yang P.
Mol Biosyst. 2010 Dec;6(12):2417-22.
A novel one-pipeline approach is reported, which can demonstrate glycoprotein identification and obtain intact glycosylation information after glycopeptide-level enrichment, without de-glycosylation. The proposed workflow has two enrichment steps plus two proteolytic processes: enriched glycoproteins were digested to peptides by Lys-C, and then enriched again and secondly digested by trypsin. In the resulting mixture, with a reasonable complexity, intact glycopeptides could be preserved and utilized informatively for glycosylation analysis, and non-glycopeptides for protein identification. In both standard protein mixture tests and real sample analysis, the resulting glycopeptides and non-glycopeptides were proved to play their expected roles, thus more confident protein glycosylation information was obtained.
Department of Chemistry, Fudan University, Shanghai 200433, P. R. China.
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