One-pipeline approach achieving glycoprotein identification and obtaining intact glycopeptide information by tandem mass spectrometry.
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.
Chen
Yaohan
Y
Department of Chemistry, Fudan University, Shanghai 200433, P. R. China.
Liu
Mingqi
M
Yan
Guoquan
G
Lu
Haojie
H
Yang
Pengyuan
P
D000595:Amino Acid Sequence; D002853:Chromatography, Liquid; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*
D006020:Glycopeptides; D006023:Glycoproteins