A mutant O-GlcNAcase as a probe to reveal global dynamics of protein O-GlcNAcylation during Drosophila embryonic development.
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.
MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Daniel
D
Mariappa
Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Nithya
N
Selvan
MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Vladimir
V
Borodkin
MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Jana
J
Alonso
MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Andrew T
AT
Ferenbach
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Claire
C
Shepherd
Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Iva Hopkins
IH
Navratilova
MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.|Division of Molecular Microbiology, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, United Kingdom.
Daan M F
DMF
vanAalten
D000215:Acylation; D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D026481:Blotting, Far-Western; D003016:Clostridium perfringens / Q000201:enzymology; D004330:Drosophila / Q000196:embryology / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D047108:Embryonic Development; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D001619:beta-N-Acetylhexosaminidases / Q000235:genetics / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D001426:Bacterial Proteins; C486300:TAB1 protein, human; C110628:hexosaminidase C; D001619:beta-N-Acetylhexosaminidases
Drosophila; Fluorescence polarization; O-GlcNAc probe; O-GlcNAcase (OGA); embryonic development