Probing the dynamics of O-GlcNAc glycosylation in the brain using quantitative proteomics

  • Khidekel N
  • Ficarro S
  • Clark P
 et al. 
  • 142

    Readers

    Mendeley users who have this article in their library.
  • 181

    Citations

    Citations of this article.

Abstract

The addition of the monosaccharide beta-N-acetyl-D-glucosamine to proteins (O-GlcNAc glycosylation) is an intracellular, post-translational modification that shares features with phosphorylation. Understanding the cellular mechanisms and signaling pathways that regulate O-GlcNAc glycosylation has been challenging because of the difficulty of detecting and quantifying the modification. Here, we describe a new strategy for monitoring the dynamics of O-GlcNAc glycosylation using quantitative mass spectrometry-based proteomics. Our method, which we have termed quantitative isotopic and chemoenzymatic tagging (QUIC-Tag), combines selective, chemoenzymatic tagging of O-GlcNAc proteins with an efficient isotopic labeling strategy. Using the method, we detect changes in O-GlcNAc glycosylation on several proteins involved in the regulation of transcription and mRNA translocation. We also provide the first evidence that O-GlcNAc glycosylation is dynamically modulated by excitatory stimulation of the brain in vivo. Finally, we use electron-transfer dissociation mass spectrometry to identify exact sites of O-GlcNAc modification. Together, our studies suggest that O-GlcNAc glycosylation occurs reversibly in neurons and, akin to phosphorylation, may have important roles in mediating the communication between neurons.

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Nelly Khidekel

  • Scott B. Ficarro

  • Peter M. Clark

  • Marian C. Bryan

  • Danielle L. Swaney

  • Jessica E. Rexach

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free