Structure-activity relationships and structural conformation of a novel urotensin II-related peptide

Urotensin II (UII) has been described as the most potent vasoconstrictor peptide and recognized as the endogenous ligand of the orphan G protein-coupled receptor GPR14. Recently, a UII-related peptide (URP) has been isolated from the rat brain and its sequence has been established as H-Ala-Cys-Phe-Trp-Lys-Tyr-Cys-Val-OH. In order to study the structure-function relationships of URP, we have synthesized a series of URP analogs and measured their binding affinity on hGPR14-transfected cells and their contractile activity in a rat aortic ring bioassay. Alanine substitution of each residue of URP significantly reduced the binding affinity and the contractile activity of the peptides, except for the Ala(8)-substituted analog that retained biological activity. Most importantly, D-scan of URP revealed that [D-Trp(4)]URP abrogated and [D-Tyr(6)]URP partially suppressed the UII-evoked contractile response. [Orn(5)]URP, which had very low agonistic efficacy, was the most potent antagonist in this series. The solution structure of URP has been determined by H-1 NMR spectroscopy and molecular dynamics. URP exhibited a single conformation characterized by an inverse gamma-turn comprising residues Trp-Lys-Tyr which plays a crucial role in the biological activity of URP. These pharmacological and structural data should prove useful for the rational design of non-peptide ligands as potential GPR14 agonists and antagonists. (C) 2004 Elsevier Inc. All rights reserved.