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Nat Biotechnol. 2016 Jan;34(1):84-8. doi: 10.1038/nbt.3403. Epub 2015 Nov 16.

Comprehensive analysis of protein glycosylation by solid-phase extraction of N-linked glycans and glycosite-containing peptides.

Author information

1
Department of Pathology, Johns Hopkins University, Baltimore, Maryland, USA.

Abstract

Comprehensive characterization of protein glycosylation is critical for understanding the structure and function of glycoproteins. However, due to the complexity and heterogeneity of glycoprotein conformations, current glycoprotein analyses focus mainly on either the de-glycosylated glycosylation site (glycosite)-containing peptides or the released glycans. Here, we describe a chemoenzymatic method called solid phase extraction of N-linked glycans and glycosite-containing peptides (NGAG) for the comprehensive characterization of glycoproteins that is able to determine glycan heterogeneity for individual glycosites in addition to providing information about the total N-linked glycan, glycosite-containing peptide and glycoprotein content of complex samples. The NGAG method can also be applied to quantitatively detect glycoprotein alterations in total and site-specific glycan occupancies.

PMID:
26571101
PMCID:
PMC4872599
DOI:
10.1038/nbt.3403
[Indexed for MEDLINE]
Free PMC Article

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