A mutant O-GlcNAcase as a probe to reveal global dynamics of protein O-GlcNAcylation during Drosophila embryonic development

Biochem J. 2015 Sep 1;470(2):255-262. doi: 10.1042/BJ20150610. Epub 2015 Jul 14.

Abstract

O-GlcNAcylation is a reversible type of serine/threonine glycosylation on nucleocytoplasmic proteins in metazoa. Various genetic approaches in several animal models have revealed that O-GlcNAcylation is essential for embryogenesis. However, the dynamic changes in global O-GlcNAcylation and the underlying mechanistic biology linking them to embryonic development is not understood. One of the limiting factors towards characterizing changes in O-GlcNAcylation has been the limited specificity of currently available tools to detect this modification. In the present study, harnessing the unusual properties of an O-GlcNAcase (OGA) mutant that binds O-GlcNAc (O-N-acetylglucosamine) sites with nanomolar affinity, we uncover changes in protein O-GlcNAcylation as a function of Drosophila development.

Keywords: Drosophila; Fluorescence polarization; O-GlcNAc probe; O-GlcNAcase (OGA); embryonic development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acylation
  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Blotting, Far-Western
  • Clostridium perfringens / enzymology
  • Drosophila / embryology
  • Drosophila / metabolism*
  • Embryo, Nonmammalian / metabolism
  • Embryonic Development
  • HEK293 Cells
  • Humans
  • Mutation
  • beta-N-Acetylhexosaminidases / genetics
  • beta-N-Acetylhexosaminidases / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Bacterial Proteins
  • TAB1 protein, human
  • hexosaminidase C
  • beta-N-Acetylhexosaminidases