Chemoproteomic discovery of ligand binding hotspots in the lipid kinome

Abstract

Diacylglycerol kinases (DGKs) are integral components of signal transduction cascades that regulate cell biology through phosphorylation of the ubiquitous secondary messenger diacylglycerol. Methods for direct evaluation of DGK activity in native biological systems are lacking and needed to develop chemical probes for studying isoform‐specific functions. Here, I will discuss efforts from my group to utilize ATP acyl phosphate activity‐based probes and quantitative mass spectrometry to map previously undefined ATP‐ and small molecule‐binding sites of representative members from all subtypes of the DGK family. We use our chemoproteomics strategy to discover an unusual binding mode for the DGK‐alpha (DGKA) inhibitor ritanserin, including interactions at a novel binding site remote from the ATP binding pocket. I will also describe our efforts towards discovery of fragment leads for future development of highly potent and selective DGKA inhibitors. Collectively, our studies illustrate the power of chemoproteomics to site‐specifically profile protein‐small molecule interactions and reveal key ligand binding sites for selective inactivation of the DGK family of lipid kinases.Support or Funding InformationThis work was supported by the University of Virginia, the LaunchPad for Diabetes Program funded by the Manning Family Foundation at the University of Virginia, National Institutes of Health Grants (DA035864 and DA043571) and U.S. Department of Defense (W81XWH‐17‐1‐0487 to K.‐L.H.).This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.