Unit

UNIT 12.8 Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine

  1. Natasha E. Zachara1,
  2. Robert N. Cole1,
  3. Gerald W. Hart1,
  4. Yuan Gao2

Published Online: 1 NOV 2001

DOI: 10.1002/0471140864.ps1208s25

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Zachara, N. E., Cole, R. N., Hart, G. W. and Gao, Y. 2001. Detection and Analysis of Proteins Modified by O-Linked N-Acetylglucosamine. Current Protocols in Protein Science. 25:12.8:12.8.1–12.8.25.

Author Information

  1. 1

    The Johns Hopkins University, Medical School, Baltimore, Maryland

  2. 2

    Deakin University, Victoria, Australia

Publication History

  1. Published Online: 1 NOV 2001
  2. Published Print: SEP 2001

This is not the most recent version of the article. View current version (1 NOV 2011)

Abstract

First, a protocol for increasing the stoichiometry of O-GlcNAc on proteins is given. This is followed by simple techniques for the detection/screening of O-GlcNAc-modified proteins either by immunoblotting or lectin affinity chromatography. Separate protocols verify that the glycan is O-linked GlcNAc. These methods are followed by protocols for more comprehensive analysis of O-GlcNAc modified proteins, including labeling of O-GlcNAc residues with [3H]Gal, and subsequent product analysis. The final two protocols assay for O-GlcNAc transferase and O-GlcNAcase activity, respectively.