Evaluation of the Chemical Reporter Analog PNP-6AzGlcNAc as an O-GlcNAcase Substrate
- Evaluation of the Chemical Reporter Analog PNP-6AzGlcNAc as an O-GlcNAcase Substrate
- 남길수; 김은주; John A Hanover; Michelle R. Bond
- Metabolic chemical reporter; O-GlcNAcase; O-GlcNAc; PNP-6AzGlcNAc
- Issue Date
- Bulletin of the Korean Chemical Society
- VOL 38, NO 2-270
- 6-Azido-6-deoxy-N-acetylglucosamine (6AzGlcNAc) was recently introduced as a new selective metabolic chemical reporter (MCR) for labeling O-GlcNAc-modified proteins in cells. To investigate whether O-6AzGlcNAc is readily cleaved by O-GlcNAcase (OGA), the enzyme responsible for removing the OGlcNAc modification, we synthesized PNP-6AzGlcNAc. This analog mimics O-GlcNAc and can be used in vitro to define the kinetic parameters for OGA thereby defining whether O-6AzGlcNAc can be employed as an appropriate tool to monitor O-GlcNAc dynamics in cells. Both PNP-6AzGlcNAc and PNP-GlcNAc were efficiently hydrolyzed by OGA with similar kinetics suggesting that an azidomodification at the GlcNAc C6 position does not significantly interfere with the β-hexosaminidase activity of OGA.
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