Affinity Purification Protocol Starting with a Small Molecule as Bait

Abstract

Protein-metabolite interactions (PMIs) are fundamental for several biological processes. Even though PMI studies have increased in recent years, our knowledge is still limited. The screening of PMIs using small molecules as bait will broaden our ability to uncover novel PMIs, setting the basis for establishing their biological relevance. Here, we describe a protocol that allows the identification of multiple protein partners for one ligand. This protocol describes a straightforward methodology that can be adapted to a wide variety of organisms.

Keywords: Affinity purification; Ligand; Mass spectrometry; Metabolites; Protein–metabolite interactions; Protein–protein interactions.