A New Searching Strategy for the Identification of O-Linked Glycopeptides

Analytical Chemistry
Jiawei MaoMingliang Ye

Abstract

For the analysis of homogeneous post-translational modifications such as protein phosphorylation and acetylation, setting a variable modification on the specific residue(s) is applied to identify the modified peptides for database searching. However, this approach is often not applicable to identify intact mucin-type O-glycopeptides due to the high microheterogeneity of the glycosylation. Because there is virtually no carbohydrate-related tag on the peptide fragments after the O-glycopeptides are dissociated in HCD, we find it is unnecessary to set the variable mass tags on the Ser/Thr residues to identify the peptide sequences. In this study, we present a novel approach, termed as O-Search, for the interpretation of O-glycopeptide HCD spectra. Instead of setting the variable mass tags on the Ser/Thr residues, we set variable mass tags on the peptide level. The precursor mass of the MS/MS spectrum was deducted by every possible summed mass of O-glycan combinations on at most three S/T residues. All the spectra with these new precursor masses were searched against the protein sequence database without setting variable glycan modifications. It was found that this method had much decreased search space and had excellent sensitivit...Continue Reading

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Citations

Jan 10, 2020·Expert Review of Proteomics·Wei-Qian CaoPeng-Yuan Yang
Feb 18, 2020·Molecular Omics·Robert J ChalkleyPeter R Baker
Sep 18, 2020·International Journal of Molecular Sciences·Xing LiXia Zou
Jul 3, 2020·Molecular & Cellular Proteomics : MCP·Weiqian CaoPengyuan Yang
Oct 28, 2020·Nature Methods·Lei LuLloyd M Smith
Feb 14, 2021·Genomics, Proteomics & Bioinformatics·Jiangming HuangWeiqian Cao
Feb 9, 2021·Molecular & Cellular Proteomics : MCP·Weiqian CaoPengyuan Yang
Jun 6, 2020·Journal of Proteome Research·Nicholas M RileyCarolyn R Bertozzi
Oct 31, 2020·Analytical Chemistry·Nicholas M RileyCarolyn R Bertozzi
Oct 19, 2019·Analytical Chemistry·Suttipong SuttapitugsakulRonghu Wu

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