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

Abstract

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 a successful glycoproteome research. In this work, we systematically investigated the behaviors and performances of boronic acid chemistry, ZIC-HILIC, and PGC of glycopeptides enrichment to promote understanding of these methods. We also optimized boronic acid chemistry and ZIC-HILIC enrichment methods and applied them to enrich intact 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 the 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 provides a better understanding of enrichment methods for further glycoproteomics research.

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Publication details

The article was received on 21 Dec 2017, accepted on 12 Feb 2018 and first published on 15 Feb 2018


Article type: Paper
DOI: 10.1039/C7AN02062G
Citation: Analyst, 2018, Accepted Manuscript
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    Study on behaviors and performances of universal N-glycopeptides enrichment methods

    Y. Xue, J. Xie, P. Fang, J. Yao, G. Yan, H. Shen and P. Yang, Analyst, 2018, Accepted Manuscript , DOI: 10.1039/C7AN02062G

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