Glycopeptide-specific monoclonal antibodies suggest new roles for O-GlcNAc

Academic Article

Abstract

  • Studies of post-translational modification by β2-N-acetyl-D- glucosamine (O-GlcNAc) are hampered by a lack of efficient tools such as O-GlcNAc-specific antibodies that can be used for detection, isolation and site localization. We have obtained a large panel of O-GlcNAc-specific IgG monoclonal antibodies having a broad spectrum of binding partners by combining three-component immunogen methodology with hybridoma technology. Immunoprecipitation followed by large-scale shotgun proteomics led to the identification of more than 200 mammalian O-GlcNAc-modified proteins, including a large number of new glycoproteins. A substantial number of the glycoproteins were enriched by only one of the antibodies. This observation, combined with the results of inhibition ELISAs, suggests that the antibodies, in addition to their O-GlcNAc dependence, also appear to have different but overlapping local peptide determinants. The monoclonal antibodies made it possible to delineate differentially modified proteins of liver in response to trauma-hemorrhage and resuscitation in a rat model.
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    Digital Object Identifier (doi)

    Author List

  • Teo CF; Ingale S; Wolfert MA; Elsayed GA; Nöt LG; Chatham JC; Wells L; Boons GJ
  • Start Page

  • 338
  • End Page

  • 343
  • Volume

  • 6
  • Issue

  • 5