Global identification of O-GlcNAc transferase (OGT) interactors by a human proteome microarray and the construction of an OGT interactome

Proteomics. 2014 May;14(9):1020-30. doi: 10.1002/pmic.201300144. Epub 2014 Mar 25.


O-Linked β-N-acetylglucosamine (O-GlcNAcylation) is an important protein PTM, which is very abundant in mammalian cells. O-GlcNAcylation is catalyzed by O-GlcNAc transferase (OGT), whose substrate specificity is believed to be regulated through interactions with other proteins. There are a handful of known human OGT interactors, which is far from enough for fully elucidating the substrate specificity of OGT. To address this challenge, we used a human proteome microarray containing ~17,000 affinity-purified human proteins to globally identify OGT interactors and identified 25 OGT-binding proteins. Bioinformatics analysis showed that these interacting proteins play a variety of roles in a wide range of cellular functions and are highly enriched in intra-Golgi vesicle-mediated transport and vitamin biosynthetic processes. Combining newly identified OGT interactors with the interactors identified prior to this study, we have constructed the first OGT interactome. Bioinformatics analysis suggests that the OGT interactome plays important roles in protein transportation/localization and transcriptional regulation. The novel OGT interactors that we identified in this study could serve as a starting point for further functional analysis. Because of its high-throughput and parallel analysis capability, we strongly believe that protein microarrays could be easily applied for the global identification of regulators for other key enzymes.

Keywords: Glycoproteomics; Human proteome microarray; O-GlcNAcylation; OGT; OGT interactome; Protein-protein interaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Glycoproteins / analysis*
  • Glycoproteins / chemistry
  • Glycoproteins / metabolism*
  • Humans
  • Immunoprecipitation
  • N-Acetylglucosaminyltransferases / chemistry
  • N-Acetylglucosaminyltransferases / metabolism*
  • Protein Array Analysis
  • Protein Interaction Maps / physiology
  • Proteome / analysis*
  • Proteome / chemistry
  • Proteome / metabolism*
  • Proteomics / methods*
  • Reproducibility of Results


  • Glycoproteins
  • Proteome
  • N-Acetylglucosaminyltransferases
  • O-GlcNAc transferase