BIOL 165 |
| In vivo carrier protein tagging has recently become an attractive target for novel site-specific fusion systems and new approaches to natural product proteomics. A detailed study of pantetheine analogs was performed in order to identify suitable partners for covalent protein labeling inside living cells. A rapid synthesis of pantethenamide analogs was developed and used to produce a panel which was evaluated for in vitro and in vivo protein labeling. Kinetic comparisons allowed the construction of a structure-activity relationship to pinpoint the linker, dye, and bioorthogonal reporter of choice for carrier protein labeling. Finally bioorthogonal pantetheine analogs were shown to target carrier protein with high specificity in vivo, and undergo chemoselective ligation to biotin reporters in crude cell lysate. The tools developed allow carrier proteins to be visualized and isolated for the first time without the expense and complication of antibody techniques |
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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 |