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O-GlcNAcase cycling: Synthesis and Applications of a novel Halotag bioprobe

Wednesday, October 10, 2012 — Poster Session II

Noon – 2:00 p.m

Natcher Conference Center, Building 45



* FARE Award Winner


  • Dona Love
  • John Hanover


Hexosamine signaling pathway is one of the key cellular responses to nutrient excess. O-GlcNAc is present on many intracellular proteins and appears to have a role in the etiology of several diseases including cancer, Alzheimer’s diseases, and type II diabetes. To monitor flux through HSP, we devised and employed a novel synthetic route to create a bioprobe specific for O-GlcNAc cycling. Our labeling strategy utilizes an engineered bacterial enzyme, haloalkane dehalogenase- the HaloTag protein, and a novel fluorogenic bioprobe substrate containing the HaloTag ligand which was recently developed by our group. Novel HaloTag ligands labeled with a small organic dye have been developed for in vivo labeling of target proteins. We designed and synthesized a potent bioprobe that is highly selective for O-GlcNAcase activity. This bioprobe contains two crucial components 1) a HaloTag reactive linker, and 2) a functional reporter such as a fluorescent dye. Here, we will leverage this technology for the specific chemical conjugation of the HaloTag ligand to a fluorogenic O-GlcNAcase-specific substrate. This bioprobe in conjunction with the HaloTag will enable us to measure the spatial and temporal activity of O-GlcNAcase as well as purify O-GlcNAc-modified targets.

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