We recently communicated a new way to study the impacts of biologically relevant reactive electrophiles on specific proteins in living cells (JACS 2013 DOI:   10.1021/ja405400). A new set of chemical tools enables spatiotemporally-controlled delivery of a given reactive entity to specific target proteins of interest (POIs) in cells  . A caged-precursor to a reactive entity covalently binds to POI (or a tag fused to POI) with 1:1 stoichiometry. At a precise time, non-intrusive photoactivation enables rapid and localized liberation of the reactive species. Quasi-intramolecular delivery design combined with electrophile-sensitivity of the POI favors rapid covalent bond-formation to the target prior to diffusion. As the only platform uniquely capable of profiling effects of non-enzyme-mediated posttranslational modifications (nE-PTMs) on one single protein, this new technology enables dissection of individual nE-PTM events dictating redox signaling. Since the strategy does not suffer from the downsides of conventional 'multi-hit' approaches wherein entire cell is bathed with reactive species, each pathway can be probed in an otherwise healthy cell. We here provide exciting results from application of this new technology in understanding the target-specific impacts of bioactive lipid-derived electrophiles on a specific gate-keeper protein regulating a major mammalian cytoprotective signaling pathway of pharmaceutical interest. Importantly, the generality of this new tool endows us and other investigators with an ability to interrogate any redox-responsive POIs and pathways they regulate with any given bioactive electrophiles of interest.

Target-specific perturbation chemistry for deconvoluting electrophile signaling biology

247th National Meeting (2014)

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