Detecting the "o-GlcNAcome"; Detection, purification, and analysis of O-GlcNAc modified proteins

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

The modification of Ser and Thr residues of cytoplasmic and nuclear proteins with a monosaccharide of O-linked β-N-acetylglucosamine is an essential and dynamic post-translational modification of metazoans. Deletion of the O-GlcNAc transferase (OGT), the enzyme that adds O-GlcNAc, is lethal in mammalian cells highlighting the importance of this post-translational modification in regulating cellular function. O-GlcNAc is believed to modulate protein function in a manner analogous to protein phosphorylation. Notably, on some proteins O-GlcNAc and O-phosphate modify the same Ser/Thr residue, suggesting that a reciprocal relationship exists between these two post-translational modifications. In this chapter we describe the most robust techniques for the detection and purification of O-GlcNAc modified proteins, and discuss some more specialized techniques for site-mapping and detection of O-GlcNAc during mass spectrometry.

Original languageEnglish (US)
Title of host publicationGlycomics
Subtitle of host publicationMethods and Protocols
EditorsNicolle Packer, Niclas Karlsson
Pages251-279
Number of pages29
DOIs
StatePublished - Dec 1 2009

Publication series

NameMethods in Molecular Biology
Volume534
ISSN (Print)1064-3745

Keywords

  • Affinity purification
  • Cellular stress
  • Mass spectrometry
  • Metabolic sensor
  • Post-translational modification
  • Signal transduction
  • Site-mapping

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Zachara, N. E. (2009). Detecting the "o-GlcNAcome"; Detection, purification, and analysis of O-GlcNAc modified proteins. In N. Packer, & N. Karlsson (Eds.), Glycomics: Methods and Protocols (pp. 251-279). (Methods in Molecular Biology; Vol. 534). https://doi.org/10.1007/978-1-59745-022-5_19