Study on behaviors and performances of universal N-glycopeptide enrichment methods

Glycosylation is a crucial process in protein biosynthesis. However, the analysis of glycopeptides through MS remains challenging due to the microheterogeneity and macroheterogeneity of the glycoprotein. Selective enrichment of glycopeptides from complex samples prior to MS analysis is essential for...

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Bibliographic Details
Published in:Analyst (London) 2018-01, Vol.143 (8), p.1870-1880
Main Authors: Xue, Yu, Xie, Juanjuan, Fang, Pan, Yao, Jun, Yan, Guoquan, Shen, Huali, Yang, Pengyuan
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibiotics
Biosynthesis
Enrichment
Glycopeptides
Glycopeptides - chemistry
Glycopeptides - isolation & purification
Glycoproteins
Glycosylation
Liver
Liver - chemistry
Male
Mannose
Mannose - chemistry
Mice
Mice, Inbred C57BL
Polysaccharides
Proteomics
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Summary:Glycosylation is a crucial process in protein biosynthesis. However, the analysis of glycopeptides through MS remains challenging due to the microheterogeneity and macroheterogeneity of the glycoprotein. Selective enrichment of glycopeptides from complex samples prior to MS analysis is essential for successful glycoproteome research. In this work, we systematically investigated the behaviors and performances of boronic acid chemistry, ZIC-HILIC, and PGC of glycopeptide enrichment to promote understanding of these methods. We also optimized boronic acid chemistry and ZIC-HILIC enrichment methods and applied them to enrich glycopeptides from mouse liver. The intact N-glycopeptides were interpreted using the in-house analysis software pGlyco 2.0. We found that boronic acid chemistry in this study preferred to capture glycopeptides with high mannose glycans, ZIC-HILIC enriched most N-glycopeptides and did not show significant preference during enrichment and PGC was not suitable for separating glycopeptides with a long amino acid sequence. We performed a detailed study on the behaviors and performances of boronic acid chemistry, ZIC-HILIC, and PGC enrichment methods and provide a better understanding of enrichment methods for further glycoproteomics research.
ISSN:0003-2654
1364-5528
DOI:10.1039/c7an02062g