Analyzing In Vivo Metabolite-Protein Interactions by Large-Scale Systematic Analyses

Metabolites interact with proteins in vivo in various ways other than enzymatic reactions. Profiling of such interactions may help disclose unknown molecular mechanisms that regulate protein functions, and provide potential targets for disease treatment. Here a procedure is described for systematic analyses of metabolite-protein interactions in vivo. This procedure couples protein affinity purification and mass spectrometry to identify metabolite-protein interactions. The primary effort can be completed within 1 day and scaled to process hundreds of samples in a batch. Originally developed in yeast, the same principles and protocol can be adapted to other organisms. Curr. Protoc. Chem. Biol. 3:181-196 © 2011 by John Wiley & Sons, Inc.

Keywords: metabolite-protein interaction; liquid chromatography; mass spectrometry; LC-MS; metabolite; protein affinity purification; yeast