Abstract:
:Compared with N-linked glycosylation, the analysis of O-GalNAc glycosylation is extremely challenging due to the high structure diversity of glycans and lack of glycosidases to release O-GalNAc glycans. In this work, a glycoform simplification strategy by combining HILIC enrichment with chemical de-sialylation to characterize O-GalNAc glycosylation of human serum is presented. This method is first validated by using the bovine fetuin as the test sample. It is found that more than 90% of the sialic acid residues can be removed from bovine fetuin by the acid-assisted de-sialylation method, which significantly simplifies the glycan structure and improves identification sensitivity. Indeed, the number of identified peptide backbones increases nearly one fold when this strategy is used. This method is further applied to analyze the human serum sample, where 185 O-GalNAc modified peptide sequences corresponding to 94 proteins with high confidence (FDR (false detection rate) <1%) are identified. This straight forward strategy can significantly reduce the variations of glycan structures, and is applicable to analysis of other biological samples with high complexity.
journal_name
Proteomicsjournal_title
Proteomicsauthors
You X,Qin H,Mao J,Tian Y,Dong M,Guo Z,Liang X,Wang L,Jin Y,Ye Mdoi
10.1002/pmic.201800042subject
Has Abstractpub_date
2018-09-01 00:00:00pages
e1800042issue
17eissn
1615-9853issn
1615-9861journal_volume
18pub_type
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