Design and Preparation of Novel Nitro-Oxide-Grafted Nanospheres with Enhanced Hydrogen Bonding Interaction for O-GlcNAc Analysis

ACS Appl Mater Interfaces. 2022 Oct 26;14(42):47482-47490. doi: 10.1021/acsami.2c15039. Epub 2022 Oct 14.


As an essential modification, O-linked β-N-acetylglucosamine (O-GlcNAc) modulates the functions of many proteins. However, site-specific characterization of O-GlcNAcylated proteins remains challenging. Herein, an innovative material grafted with nitro-oxide (N→O) groups was designed for high affinity enrichment for O-GlcNAc peptides from native proteins. By testing with synthetic O-GlcNAc peptides and standard proteins, the synthesized material exhibited high affinity and selectivity. Based on the material prepared, we developed a workflow for site-specific analysis of O-GlcNAcylated proteins in complex samples. We performed O-GlcNAc proteomics with the PANC-1 cell line, a representative model for pancreatic ductal adenocarcinoma. In total 364 O-GlcNAc peptides from 267 proteins were identified from PANC-1 cells. Among them, 183 proteins were newly found to be O-GlcNAcylated in humans (with 197 O-GlcNAc sites newly reported). The materials and methods can be facilely applied for site-specific O-GlcNAc proteomics in other complex samples.

Keywords: O-GlcNAc; O-GlcNAcylation; glycopeptide enrichment; glycoproteomics; hydrogen bonding interaction; nitro-oxide-grafted nanosphere.

MeSH terms

  • Acetylglucosamine* / analysis
  • Acetylglucosamine* / chemistry
  • Acetylglucosamine* / metabolism
  • Humans
  • Hydrogen Bonding
  • Nanospheres*
  • Oxides
  • Peptides
  • Proteins


  • Acetylglucosamine
  • Oxides
  • Proteins
  • Peptides