RT Journal Article SR Electronic T1 The Use of Bioluminescence Resonance Energy Transfer 2 to Study Neuropeptide Y Receptor Agonist-Induced β-Arrestin 2 Interaction JF Journal of Pharmacology and Experimental Therapeutics JO J Pharmacol Exp Ther FD American Society for Pharmacology and Experimental Therapeutics SP 147 OP 156 DO 10.1124/jpet.103.051227 VO 306 IS 1 A1 Magnus M. Berglund A1 Douglas A. Schober A1 Michael A. Statnick A1 Patricia H. McDonald A1 Donald R. Gehlert YR 2003 UL http://jpet.aspetjournals.org/content/306/1/147.abstract AB The neuropeptide Y (NPY) family peptides NPY, peptide YY (PYY), and pancreatic polypeptide (PP) bind to four G protein-coupled receptors (GPCRs): Y1, Y2, Y4, and Y5. A key step in the desensitization and internalization of GPCRs is the association of the receptor with β-arrestins. In the present study, these receptors were analyzed with respect to their ability to interact with GFP2-tagged β-arrestin 2 using the new bioluminescence resonance energy transfer 2 method. Agonists induced a concentration-dependent association of β-arrestin 2 with all four receptors. Whereas the Y1 receptor exhibited the highest maximum response and rapid association (t½ = 3.4 min), the maximal signals for the association of Y2 and Y4 receptors were less than half of that of Y1, and the association rates were much slower. Interestingly, when evaluated at the Y4 receptor, the Y4 agonist 1229U91 [(Ile,Glu,Pro,Dpr,Tyr,Arg, Leu,Arg,Try-NH2)-2-cyclic(2,4′),(2′,4)-diamide] was unable to provoke the same maximal response as human PP, suggesting that 1229U91 is a partial agonist. When stimulated by PYY, the Y5 receptor responded with a t½ of 4.6 min and a maximal response approximately 60% of what was observed with Y1. Because β-arrestins are key components in GPCR internalization, it is interesting to note that the receptor that is known to internalize rapidly (Y1) exhibits the most rapid association with β-arrestin 2, whereas the receptor that is known to internalize slowly, or not at all (Y2) associates slowly with β-arrestin 2. The American Society for Pharmacology and Experimental Therapeutics