BIOL 110 |
| We are developing a high-throughput assay capable of dissecting the molecular interactions of glycosylated proteins. This assay was inspired by the traditional yeast two-hybrid assay, which is an extremely powerful genetic technique for the detection of protein-protein interactions. The Golgi two-hybrid will test protein-protein interactions in the late compartments of the secretory pathway, where these important proteins will have their full complement of post-translational modifications. Our assay takes advantage of the fact that Golgi-resident glycosyltransferases can be subdivided into modular domains that can be reassembled. The modular glycosyltransferase domains will be fused to the “bait” (protein of interest) and “prey” (library of potential interaction partners). If the bait interacts with a prey protein within the selection cell line, the glycosyltransferase will become active and will serve as a reporter of the interaction. Here we evaluate possible ways to link protein-protein interactions to glycosyltransferase activity via a robust selection strategy. |
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Chemistry and Metabolism
4:30 PM-6:30 PM, Tuesday, 12 September 2006 Moscone Center -- Hall D, Poster
Division of Biological Chemistry |