Mapping the oxysterol interactome

Abstract

Oxysterols are signaling mols. that translate metabolic information to cellular response. In mammalian cells, oxysterols serve essential roles in transcriptional regulation, membrane integrity, and the activity of target proteins. Accordingly, oxysterols coordinate diverse events that include cholesterol biosynthesis, immune response, and viral infection and defense. Despite their potent effects, the structural code that dets. the specific activity of oxysterols remains uncharacterized. We have created a unique oxysterol scaffold that preserves the biophys. properties of the natural mols. and contains functional groups to covalently capture and identify cellular interactors. We describe the synthesis and characterization of one such mol. and demonstrate its capacity to recapitulate the activity of its natural congener within the Hedgehog signaling pathway. Our detailed biochem., transcriptional, and chemoproteomic studies define these probes as powerful tools to reveal how Nature templates function through metabolite structure.