Direct Proximity Tagging of Small Molecule Protein Targets Using an Engineered NEDD8 Ligase.

ABSTRACT: Identifying the protein targets of bioactive small molecules remains a major problem in the discovery of new chemical probes and therapeutics. While activity-based probes and photo-cross-linkers have had success in identifying protein targets of small molecules, each technique has limitations. Here we describe a method for direct proximity tagging of proteins that bind small molecules. We engineered a promiscuous ligase based on the NEDD8 conjugating enzyme, Ubc12, which can be covalently linked to a small molecule of interest. When target proteins bind the small molecule, they are directly labeled on surface lysines with a biotinylated derivative of the small ubiquitin homologue, NEDD8. This unique covalent tag can then be used to identify the small molecule binding proteins. Utilizing th

SUBMITTER: Hill ZB 

PROVIDER: S-EPMC5308480 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature