C5a-stimulated recruitment of beta-arrestin2 to the nonsignaling 7-transmembrane decoy receptor C5L2.

C5L2 (or GPR77) is a high-affinity receptor for the complement fragment C5a and its desarginated product, C5a-desArg. Unlike the classical C5a receptor CD88, C5L2 does not couple to intracellular G-protein-signaling pathways but is thought to function as a decoy receptor. The authors show that stimulation of C5L2 with C5a and C5a-desArg induces redistribution of green fluorescent protein-labeled beta-arrestin2 to cytoplasmic vesicles. C3a and C3a-desArg were inactive in the beta-arrestin translocation assay. Direct interaction of ligand-stimulated C5L2 with beta-arrestin was confirmed using a novel beta-galactosidase fragment complementation assay. In this assay, C5L2 was labeled with a mutationally altered peptide fragment of beta-galactosidase, whereas beta-arrestin2 was labeled with a corresponding deletion mutant of the enzyme. Stable transfection of the modified C5L2 and subsequent stimulation with C5a or C5a-desArg restored beta-galactosidase activity in a dose-dependent manner. The subnanomolar potency of beta-arrestin coupling in the beta-galactosidase fragment complementation assay is in agreement with the affinity of the receptor-ligand interaction. C5L2 is the first example of a 7-transmembrane decoy receptor that couples to beta-arrestin in a ligand-dependent manner. This observation supports the notion that G-protein-signaling and beta-arrestin coupling can be 2 separate activities of 7-transmembrane receptors. Furthermore, the beta-arrestin assays described in this article provide methods of screening for selective C5L2 modulators.