Glyco-DIA: a method for quantitative O-glycoproteomics with in silico-boosted glycopeptide libraries

TY - JOUR

T1 - Glyco-DIA

T2 - a method for quantitative O-glycoproteomics with in silico-boosted glycopeptide libraries

AU - Ye, Zilu

AU - Mao, Yang

AU - Clausen, Henrik

AU - Vakhrushev, Sergey Y

PY - 2019/9/1

Y1 - 2019/9/1

N2 - We report a liquid chromatography coupled to tandem mass spectrometry O-glycoproteomics strategy using data-independent acquisition (DIA) mode for direct analysis of O-glycoproteins. This approach enables characterization of glycopeptides and structures of O-glycans on a proteome-wide scale with quantification of stoichiometries (though it does not allow for direct unambiguous glycosite identification). The method relies on a spectral library of O-glycopeptides; the Glyco-DIA library contains sublibraries obtained from human cell lines and human serum, and it currently covers 2,076 O-glycoproteins (11,452 unique glycopeptide sequences) and the 5 most common core1 O-glycan structures. Applying the Glyco-DIA library to human serum without enrichment for glycopeptides enabled us to identify and quantify 269 distinct glycopeptide sequences bearing up to 5 different core1 O-glycans from 159 glycoproteins in a SingleShot analysis.

AB - We report a liquid chromatography coupled to tandem mass spectrometry O-glycoproteomics strategy using data-independent acquisition (DIA) mode for direct analysis of O-glycoproteins. This approach enables characterization of glycopeptides and structures of O-glycans on a proteome-wide scale with quantification of stoichiometries (though it does not allow for direct unambiguous glycosite identification). The method relies on a spectral library of O-glycopeptides; the Glyco-DIA library contains sublibraries obtained from human cell lines and human serum, and it currently covers 2,076 O-glycoproteins (11,452 unique glycopeptide sequences) and the 5 most common core1 O-glycan structures. Applying the Glyco-DIA library to human serum without enrichment for glycopeptides enabled us to identify and quantify 269 distinct glycopeptide sequences bearing up to 5 different core1 O-glycans from 159 glycoproteins in a SingleShot analysis.

U2 - 10.1038/s41592-019-0504-x

DO - 10.1038/s41592-019-0504-x

M3 - Journal article

C2 - 31384044

SN - 1548-7091

VL - 16

SP - 902

EP - 910

JO - Nature Methods

JF - Nature Methods

IS - 9

ER -