α-Methylene-β-Lactone Scaffold for Developing Chemical Probes at the Two Ends of the Selectivity Spectrum |
| |
Authors: | Dr Lei Wang Louis P Riel Dr Bekim Bajrami Prof?Dr Bin Deng Prof?Dr Amy R Howell Prof?Dr Xudong Yao |
| |
Affiliation: | 1. Department of Chemistry, University of Connecticut, Storrs, CT 06269 USA;2. Chemical Biology & Proteomics, Biogen, Cambridge, MA 02142 USA;3. Department of Biology, University of Vermont, Burlington, VT 05405 USA
Vermont Genetics Network Proteomics Facility, University of Vermont, Burlington, VT 05405 USA |
| |
Abstract: | The utilities of an α-methylene-β-lactone (MeLac) moiety as a warhead composed of multiple electrophilic sites are reported. We demonstrate that a MeLac-alkyne not only reacts with diverse proteins as a broadly reactive measurement probe, but also recruits reduced endogenous glutathione (GSH) to assemble a selective chemical probe of GSH-β-lactone (GSH-Lac)-alkyne in live cells. Tandem mass spectrometry reveals that MeLac reacts with nucleophilic cysteine, serine, lysine, threonine, and tyrosine residues, through either Michael or acyl addition. A peptide-centric proteomics platform demonstrates that the proteomic selectivity profiles of orlistat and parthenolide, which have distinct reactivities, are measurable by MeLac-alkyne as a high-coverage probe. The GSH-Lac-alkyne selectively probes the glutathione S-transferase P responsible for multidrug resistance. The assembly of the GSH-Lac probe exemplifies a modular and scalable route to develop selective probes with different recognizing moieties. |
| |
Keywords: | chemical proteomics glutathione S-transferase methylenelactone orlistat parthenolide |
|
|