Structure–activity relationships and structural conformation of a novel urotensin II-related peptide☆
Introduction
Urotensin II (UII) is a cyclic peptide that has been originally isolated from the caudal neurosecretory organ (termed urophysis) of teleost fish [53]. Characterization of UII from the brain of a frog has subsequently shown that a UII gene is also present in tetrapods and that this gene is expressed in the CNS [14]. In fact, the cDNA, encoding the UII precursor, has now been cloned in various species of vertebrates including carp [51], frog [16], mouse [17], rat [17], pig [47], monkey [21] and human [16]. The structure of the cyclic hexapeptide located in the C-terminal portion of UII has been fully conserved from fish to mammals while the sequence of the N-terminal region is highly variable (Fig. 1).
Pharmacological studies have shown that UII exercices a large array of spasmogenic actions. Notably, in fish and amphibians, UII provokes smooth muscle contraction in various vascular, intestinal and urogenital tissues [15]. In mammals, UII exerts a complex myotropic action on smooth muscles. Early studies have shown that UII produces relaxation of the mouse anococcygeus muscle [25] and causes hypotension in anesthetized rat [26]. More recently, UII has been found to exert potent contractile activity in rat, porcine, dog and primate vascular tissues [2], [8], [19] and primate airway smooth muscles [29]. In addition, intracerebroventricular or intracerebral administration of UII elicits various cardiovascular [36], [41], endocrine [24] and behavioral effects [24].
Using a reverse pharmacology approach, four groups have concomitantly shown that UII is the cognate ligand for the orphan G protein-coupled receptor GPR14 [2], [40], [47], [50]. The UII receptor is widely expressed in various peripheral organs including the cardiovascular system, the kidney, the bladder, as well as the central nervous system [2], [11], [24], [40], [44], [61], [63]. The variety of effects of UII and the broad expression pattern of its receptors indicate that UII may be involved in physiopathological processes. Indeed, recent studies suggest that UII may play a role in smooth muscle cell proliferation [56], [66], cardiac fibrosis and hypertrophy [64], [70], heart failure [66], cardiac remodeling [6], [20], atherosclerosis [62] and renal dysfunction [63].
A recent report has demonstrated the existence, in mouse, rat and human, of a paralog of UII named UII-related peptide (URP) [60] (Fig. 1). In the rat brain, the URP isoform has been purified and sequenced whereas the UII isoform has not yet been isolated [60]. Further, synthetic URP exhibits a higher binding affinity and a slightly higher potency to increase Ca2+ mobilization in human GPR14-transfected cells than the UII variant [60], suggesting that the biological effects previously attributed to UII could actually be exerted, at least in part, by its paralog URP.
In the present report, we have studied the structure–activity relationships of a series of URP analogs using a radioligand binding assay with hGPR14-transfected cells and an aortic ring contraction assay, and we have determined the solution conformation of URP by 1H NMR and molecular dynamics.
Section snippets
Chemicals and reagents
All l-amino acid residues, preloaded polyethylene glycol–polystyrene resins (Fmoc-Val-PEG-PS, Fmoc-Ala-PEG-PS), dichloromethane (DCM), 1-hydroxybenzotriazole (HOBt) and trifluoroacetic acid (TFA) were purchased from Applera-France (Courtaboeuf, France). Fmoc-3-iodo-Tyr-OH, Fmoc-Cys(Acm)-OH and all d-amino acid residues were obtained from Bachem Biochimie (Weil am Rhein, Germany). Fmoc-Pen(Acm)-OH was from VWR (Fontenay-sous-Bois, France). N,N-dimethylformamide (DMF), isopropanol (iPrOH) and
Peptide analysis
RP-HPLC analysis of hUII, URP and URP analogs revealed that the purity of all peptides was higher than 98% (Table 1). For all peptides, the molecular weight observed by MS analysis agreed with the theoretical values (Table 1). These physicochemical data ensured that pharmacological and functional information as well as structural parameters were undoubtedly assigned to the considered analog.
Structure–activity relationships
The binding affinity of URP (compound 1) on human GPR14-transfected cells was slightly higher than that
Discussion
Soon after the UII precursor had been characterized in human [16], several studies have shown that UII is a natural ligand of the orphan receptor GPR14 [2], [40], [47], [50]. Recently, a UII-related peptide called URP has been isolated from the rat brain [60] and the URP precursor cDNA has been cloned in human, rat and mouse [60]. Binding and functional studies have revealed that URP, like UII, is a potent agonist of GPR14 [60]. In the present report, we describe the structure–activity
Acknowledgments
This study was supported by INSERM and the Ministère de la Jeunesse, de l’Education Nationale et de la Recherche. D.C. and C.D. were recipients of fellowships from INSERM-SERVIER laboratories.
References (70)
- et al.
NMR and dynamical simulated annealing studies on the solution conformation of urotensin II
Biochem Biophys Acta
(1994) - et al.
The urotensin II receptor is expressed in the cholinergic mesopontine tegmentum of the rat
Brain Res
(2001) - et al.
Post-translational processing of prepro-urotensin II
FEBS Lett
(1990) - et al.
Isolation and primary structure of urotensin II from the brain of a tetrapod, the frog Rana ridibunda
Biochem Biophys Res Commun
(1992) - et al.
Cloning, sequence analysis and tissue distribution of the mouse and rat urotensin II precursors
FEBS Lett
(1999) - et al.
Structural requirements at the N-terminus of urotensin II octapeptides
Peptides
(2002) - et al.
Congestive heart failure and expression of myocardial urotensin II
Lancet
(2002) - et al.
Cardiovascular effects of urotensin II in anesthetized and pithed rats
Gen Comp Neurol
(1986) - et al.
Design, synthesis, conformational analysis, and biological studies of urotensin-II lactam analogues
Bioorg Med Chem
(2002) - et al.
Primary structures of multiple forms of urotensin II in the urophysis of the carp Cyprinus Carpio
Gen Comp Endocrinol
(1984)
Identification of urotensin II as the endogenous ligand for the orphan G-protein-coupled receptor GPR14
Biochem Biophys Res Commun
Cardiovascular effects of urotensin II in different brain areas
Peptides
Cloning and chromosomal mapping of three novel genes, GPR9, GPR10, and GPR14, encoding receptors related to interleukin 8, neuropeptide Y, and somatostatin receptors
Genomics
Isolation and amino acid sequence of two urotensin II peptides from Catostomus commersoni urophyses
Peptides
Isolation and amino acid sequence of urotensin II from the sturgeon Acipenser ruthenus
Gen Comp Endocrinol
Urotensin II is the endogenous ligand of a G-protein-coupled orphan receptor, SENR (GPR14)
Biochem Biophys Res Commun
Determination of three-dimensional structures of proteins from interproton distance data by hybrid distance geometry-dynamical simulated annealing calculations
FEBS Lett
Calibration of the angular dependence of the amide proton–alpha proton coupling constants, 3JHN alpha, in a globular proteinUse of 3JHN alpha for identification of helical secondary structure
J Mol Biol
Turns in peptides and proteins
Adv Protein Chem
An improved method for anchoring of 9-fluorenylmethoxycarbonyl-amino acids to 4-alkoxybenzylalcohol resins
Tetrahedron Lett
Gradient-tailored water suppression for 1H–15N HSQC experiments optimized to retain full sensitivity
J Magn Reson
Identification of urotensin II-related peptide as the urotensin II-immunoreactive molecule in the rat brain
Biochem Biophys Res Commun
A novel putative neuropeptide receptor expressed in neural tissue, including sensory epithelia
Biochem Biophys Res Commun
Urotensin II-induced activation of extracellular signal-regulated kinase in cultured vascular smooth muscle cells: involvement of cell adhesion-mediated integrin signaling
Life Sci
Role of urotensin II in patients on dialysis
Lancet
Purification and characterization of urotensin II from the brain of a teleost (trout, Oncorhynchus mykiss) and an elasmobranch (skate, Raja rhina)
Gen Comp Endocrinol
Urotensin II from the river lamprey (Lampetra fluviatilis), the sea lamprey (Petromyzon marinus), and the paddlefish (Polyodon spathula)
Gen Comp Endocrinol
Urotensin II induces hypertrophic responses in cultured cardiomyocytes from neonatal rats
FEBS Lett
Cyclization of disulfide-containing peptides in solid-phase synthesis
Int J Pept Protein Res
Human urotensin-II is a potent vasoconstrictor and agonist for the orphan receptor GPR14
Nature
Comparative distribution of pituitary adenylate cyclase-activating polypeptide (PACAP) binding sites and PACAP receptor mRNAs in the rat brain during development
J Comp Neurol
Neuropeptide Y: identification of the binding site
Int J Pept Protein Res
Pharmacological characterization of SB-710411 (Cpa-c[d-Cys-Pal-d-Trp-Lys-Val-Cys]-Cpa-amide), a novel peptidic urotensin-II receptor antagonist
Br J Pharmacol
Deletion of the UT receptor gene results in the selective loss of urotensin-II contractile activity in aortae isolated from UT receptor knockout mice
Br J Pharmacol
Effects of human urotensin II in isolated vessels of various species: comparison with other vasoactive agents
Naunyn Schmiedebergs Arch Pharmacol
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Identification and signaling characterization of four urotensin II receptor subtypes in the western clawed frog, Xenopus tropicalis
2020, General and Comparative EndocrinologyCitation Excerpt :A paralogous peptide, termed UII-related peptide (URP), was later reported in rodents and humans (Sugo et al., 2003), birds (Tostivint et al., 2006), Japanese eel (Nobata et al., 2011), and African clawed frog (Konno et al., 2013). The C-terminal hexapeptide (CFWKYC) of UII and URP are highly conserved among vertebrates (Vaudry et al., 2010) and exhibits full biological activity (Chatenet et al., 2004; Douglas and Ohlstein, 2000; Leprince et al., 2008). UII and URP act via G-protein-coupled receptor 14 (GPR14), now termed the urotensin II receptor (UTR) (Vaudry et al., 2015).
Endozepines and their receptors: Structure, functions and pathophysiological significance
2020, Pharmacology and TherapeuticsMicroinjection of urotensin II into the pedunculopontine tegmentum leads to an increase in the consumption of sweet tastants
2020, Physiology and BehaviorCitation Excerpt :Daily handling continued throughout the study. Although the endogenous activating ligand for UIIRs expressed in the supraspinal CNS is likely the urotensin II-related peptide (URP), our experiments use UII for the following reasons, the peptides: (1) have the same affinity and potency [46,47], (2) have the same core chemophore [46], and (3) to put our results into context of previous literature which exclusively uses UII. UII (Kinexus Bioinformatics, Vancouver, BC) was prepared in artificial cerebrospinal fluid (aCSF) and 0.01% bovine serum albumin at a concentration of 10 µM.
Towards Targeting the Urotensinergic System: Overview and Challenges
2019, Trends in Pharmacological Sciences
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Symbols and abbreviations are in accord with the recommendations of the IUPAC-IUB Joint Commission on Biochemical Nomenclature: Nomenclature and Symbolism for Amino Acids and Peptides, Biochem. J. 1984, 219, 345–373, and of A Short Guide to Abbreviations and Their Use in Peptide Science, J. Peptide Sci. 2003, 9, 1–8. Additional abbreviations are as follows: RP-HPLC, reversed-phase high-performance liquid chromatography; NMR, nuclear magnetic resonance; MALDI-TOF-MS, matrix-assisted laser desorption ionization-time of flight mass spectrometry. All optically active amino acids are of the l-configuration unless otherwise noted.
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Authors who equally contributed to this work.