Abstract：
:Confident characterization of the microheterogeneity of protein glycosylation through identification of intact glycopeptides remains one of the toughest analytical challenges for glycoproteomics. Recently proposed mass spectrometry (MS)-based methods still have some defects such as lack of the false discovery rate (FDR) analysis for the glycan identification and lack of sufficient fragmentation information for the peptide identification. Here we proposed pGlyco, a novel pipeline for the identification of intact glycopeptides by using complementary MS techniques: 1) HCD-MS/MS followed by product-dependent CID-MS/MS was used to provide complementary fragments to identify the glycans, and a novel target-decoy method was developed to estimate the false discovery rate of the glycan identification; 2) data-dependent acquisition of MS3 for some most intense peaks of HCD-MS/MS was used to provide fragments to identify the peptide backbones. By integrating HCD-MS/MS, CID-MS/MS and MS3, intact glycopeptides could be confidently identified. With pGlyco, a standard glycoprotein mixture was analyzed in the Orbitrap Fusion, and 309 non-redundant intact glycopeptides were identified with detailed spectral information of both glycans and peptides.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Zeng WF,Liu MQ,Zhang Y,Wu JQ,Fang P,Peng C,Nie A,Yan G,Cao W,Liu C,Chi H,Sun RX,Wong CC,He SM,Yang Pdoi
10.1038/srep25102subject
Has Abstractpub_date
2016-05-03 00:00:00pages
25102issn
2045-2322pii
srep25102journal_volume
6pub_type
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