Selective Orthosteric Free Fatty Acid Receptor 2 (FFA2) Agonists

• DOI: 10.1074/jbc.m110.210872
• Publication Date: 2011-01-11T03:54:56Z
• Authors: Johannes Schmidt; Nicola J. Smith; Elisabeth Christi...; Irina G. Tikhonova; Manuel Grundmann; Brian D. Hudson; Richard J. Ward; Christel Drewke; Graeme Milligan; Evi Kostenis; Trond Ulven
• Keywords: /dk/atira/pure/subjectarea/asjc/1300/1303; 0303 health sciences; /dk/atira/pure/subjectarea/asjc/1300/1312; name=Molecular Biology; Binding Sites; /dk/atira/pure/subjectarea/asjc/1300/1312; name=Biochemistry; Receptors, Cell Surface; name=Molecular Biology; 540; Ligands; name=Cell Biology; Receptors, G-Protein-Coupled; /dk/atira/pure/subjectarea/asjc/1300/1307; name=Cell Biology; Structure-Activity Relationship; 03 medical and health sciences; /dk/atira/pure/subjectarea/asjc/1300/1303; name=Biochemistry; HEK293 Cells; 615; Humans; Propionates; /dk/atira/pure/subjectarea/asjc/1300/1307; Signal Transduction

Free fatty acid receptor 2 (FFA2; GPR43) is a G protein-coupled seven-transmembrane for short-chain acids (SCFAs) that implicated in inflammatory and metabolic disorders. The SCFA propionate has close to optimal ligand efficiency FFA2 can hence be considered as highly potent given its size. Propionate, however, does not discriminate between the closely related FFA3 (GPR41). To identify FFA2-selective ligands understand molecular basis selectivity, targeted library of small carboxylic was examined using holistic, label-free dynamic mass redistribution technology primary screening receptor-proximal protein [(35)S]guanosine 5'-(3-O-thio)triphosphate activation, inositol phosphate, cAMP accumulation assays hit confirmation. Structure-activity relationship analysis allowed formulation general rule predict selectivity at orthosteric binding site where with substituted sp(3)-hybridized α-carbons preferentially activate FFA3, whereas sp(2)- or sp-hybridized prefer FFA2. mode verified by site-directed mutagenesis: replacement arginine residues alanine prevented binding, modeling predicted detailed binding. Based on this, selective mutation three their non-conserved counterparts sufficient transfer Thus, activation via achievable rather ligands, finding significant implications rational design therapeutic compounds selectively targeting receptors.