Recognition of Core-Fucosylated Glycopeptides Based on the Y1+Fuc/Y1 Ratio in Low-Energy HCD Spectra

Anal Chem. 2022 Dec 20;94(50):17349-17353. doi: 10.1021/acs.analchem.2c03182. Epub 2022 Dec 9.

Abstract

Accurate identification of core fucosylation on N-glycopeptides remains challenging due to fucose migration during mass spectrometry analysis. Here, we introduce a simple and straightforward method for core-fucosylated glycopeptide recognition based on the relative intensities of Y1+Fuc ions compared with their corresponding Y1 ions (labeled as Y1+Fuc/Y1 or simply Y1F/Y1 ratio > 0.1) in low-energy HCD-based spectra. The method was first developed by systematically evaluating the influence of fucose migration on the Y1F ion from antenna fucoses based on the distribution of the Y1F/Y1 ratios in the MS/MS spectra of antenna-fucosylated glycopeptides from Fut8-/- mouse brain. The feasibility of the method was then confirmed by using two standard glycoproteins, comparison with glycopeptides in Fut8+/+ mouse brain with/without in silico core-fucosylation removal, and Y1F/Y1 ratio alterations under a lower HCD energy. This method will be applicable to the manual interpretation and software-based high-throughput analysis of core-fucosylated glycopeptides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fucose / chemistry
  • Glycopeptides* / analysis
  • Glycoproteins / chemistry
  • Glycosylation
  • Mice
  • Tandem Mass Spectrometry* / methods

Substances

  • Glycopeptides
  • Fucose
  • Glycoproteins