Location of Repository

Human urotensin-II receptor desensitisation

By Madura Suharshana Batuwangala


Human Urotensin-II (U-II) is a cyclic undecapeptide that binds to the U-II receptor UT. The desensitisation mechanisms of the UT receptor (G_{q/11} coupled GPCR) are not well defined and hampered by (1) lack of native (in-vitro) models; (2) paucity of ligands, especially non-peptides and (3) irreversible binding of U-II. There are some limited studies using rat aorta, where a U-II induced primary contractile response was reduced upon a secondary re-challenge after 5-hours. \ud Studies were undertaken to characterise cell lines expressing native (SJCRH30) and recombinant human hUT (HEK293 and CHO) for their suitability in binding and functional assays (PI and Ca^2+). SAR studies were carried out to characterise novel analogues modified at Tyr^9 of the U-II(4-11) template. This led to the identification of [3,5-diiodoTyr^9]U-II(4-11) a partial agonist in aorta and Ca^2+ assays at rat UT. Full agonism was demonstrated at hUT in PI and Ca^2+ assays. Efforts were made to delineate functional and genomic desensitisation of hUT. There was no functional desensitisation in SJCRH30. In HEK293hUT functional heterologous desensitisation of hUT was observed, this was not so in CHOhUT; instead P_2YR was functionally attenuated. In SJCRH30 6-hr U-II treatments led to UT mRNA reduction. Genomic desensitisation was also studied in Peripheral blood mononuclear cells (PBMCs). U-II treatments alone did not affect UT mRNA. Lipolysaccharide treatment of PBMCs led to UT mRNA upregulation which was desensitised with U-II treatments. In recombinant systems UT mRNA was upregulated at 6-hr U-II treatments. \ud In conclusion modification of the U-II(4-11) template at Tyr^9 is useful for reducing efficacy. There is a difference in desensitisation profiles of native and recombinant hUT, where native receptors are not prone to functional desensitisation while receptor mRNA is reduced. In recombinant systems, hUT undergoes desensitisation (HEK293hUT only) while receptor mRNA is increased in both systems

Publisher: University of Leicester
Year: 2009
OAI identifier: oai:lra.le.ac.uk:2381/7846

Suggested articles



  1. (1979). [1-Penicillamine,2-leucine]oxytocin. Synthesis and pharmacological and conformational studies of a potent peptide hormone inhibitor.
  2. (2004). A
  3. (2005). A Guide to wire myography.
  4. (1985). A new generation of Ca2+ indicators with greatly improved fluorescence properties.
  5. (2002). A new ligand for the urotensin II receptor.
  6. (2002). A new, potent urotensin II receptor peptide agonist containing a Pen residue at the disulfide bridge.
  7. (2009). A Pharmacology Primer: Theory, Application and Methods. Third edn.
  8. (2006). A signature pattern of stress-responsive microRNAs that can evoke cardiac hypertrophy and heart failure.
  9. (1998). A tool coming of age: thapsigargin as an inhibitor of sarco-endoplasmic reticulum Ca(2+)-ATPases. Trends Pharmacol
  10. (1998). Activation of stressactivated MAP protein kinases up-regulates expression of transgenes driven by the cytomegalovirus immediate/early promoter.
  11. (1999). Activators and target genes of Rel/NF-kappaB transcription factors.
  12. (2008). Agonist-selective mechanisms of GPCR desensitization.
  13. (1981). alpha-adrenergic antagonists as possible calcium channel inhibitors.
  14. (2006). Alteration of vascular urotensin II receptor in mice with apolipoprotein E gene knockout.
  15. (2007). An Introduction to real-time PCR. In: Dorak,
  16. (2001). Analysis of relative gene expression data using realtime quantitative PCR and the 2(-Delta Delta C(T)) Method.
  17. (2005). Architecture of the human urotensin II receptor: comparison of the binding domains of peptide and non-peptide urotensin II agonists.
  18. (2002). Are we beta-ARKing up the wrong tree? Casein kinase 1 alpha provides an additional pathway for GPCR phosphorylation.
  19. (2006). Arrestins: ubiquitous regulators of cellular signaling pathways.
  20. (2006). Arsenite induces apoptosis in hepatocytes through an enhancement of the activation of Jun N-terminal kinase and p38 mitogen-activated protein kinase caused by partial hepatectomy.
  21. (2005). Behavioral effects of urotensin-II centrally administered in mice.
  22. (1990). beta-Arrestin: a protein that regulates beta-adrenergic receptor function.
  23. (2008). Bi-directional heterologous desensitization between the major HIV-1 co-receptor CXCR4 and the kappa-opioid receptor.
  24. (2000). Bidirectional heterologous desensitization of opioid and chemokine receptors.
  25. (2008). Biological properties and functional determinants of the urotensin II receptor.
  26. (1993). Ca2+ limits the development of the light response in Drosophila photoreceptors.
  27. (2008). Ca2+-operated transcriptional networks: molecular mechanisms and in vivo models.
  28. (2001). Cardiostimulant effects of urotensin-II in human heart in vitro.
  29. (1983). Catecholamine-induced desensitization of turkey erythrocyte adenylate cyclase is associated with phosphorylation of the beta-adrenergic receptor.
  30. (2006). Cell and tissue responses of a range of Urotensin II analogs at cloned and native urotensin II receptors. Evidence for coupling promiscuity.
  31. (2003). Central cardiovascular action of urotensin II in spontaneously hypertensive rats.
  32. (2004). Central effects of native urotensin II on motor activity, ventilatory movements, and heart rate in the trout Oncorhynchus mykiss.
  33. (2001). Central effects of urotensin-II following ICV administration in rats.
  34. (2008). Chronic urotensin II infusion enhances macrophage foam cell formation and atherosclerosis in apolipoprotein E-knockout mice.
  35. (2006). Circulating urotensin II levels in moderate to severe congestive heart failure: its relations with myocardial function and well established neurohormonal markers.
  36. (1995). Cloning and chromosomal mapping of three novel genes, GPR9, GPR10, and GPR14, encoding receptors related to interleukin 8, neuropeptide Y, and somatostatin receptors.
  37. (1998). Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord.
  38. (1999). Cloning, sequence analysis and tissue distribution of the mouse and rat urotensin II precursors.
  39. (1995). Co-expression in CHO cells of two muscle proteins involved in excitation-contraction coupling.
  40. (1999). Comparison of SYBR Green I nucleic acid gel stain mutagenicity and ethidium bromide mutagenicity in the Salmonella/mammalian microsome reverse mutation assay (Ames test).
  41. (2006). Confocal microscopy: theory and applications for cellular signaling In: Lambert, D (ed). Calcium Signaling Protocols.
  42. CR (2001a). Synergistic effect of urotensin II with mildly oxidized LDL on DNA synthesis in vascular smooth muscle cells.
  43. CR (2001b). Synergistic effect of urotensin II with serotonin on vascular smooth muscle cell proliferation.
  44. (1996). Cross-desensitization among receptors for platelet activating factor and peptide chemoattractants. Evidence for independent regulatory pathways.
  45. D (2002b). Effects of human urotensin II in isolated vessels of various species; comparison with other vasoactive agents.
  46. (2007). Definition of new pharmacophores for nonpeptide antagonists of human urotensin-II. Comparison with the 3D-structure of human urotensin-II and URP.
  47. (2003). Deletion of the UT receptor gene results in the selective loss of urotensin-II contractile activity in aortae isolated from UT receptor knockout mice.
  48. (1999). Desensitization of G-protein-coupled receptors in the cardiovascular system.
  49. (1994). Desensitization of neuromedin B receptors (NMB-R) on native and NMB-R-transfected cells involves downregulation and internalization.
  50. (1992). Desensitization of the isolated beta 2-adrenergic receptor by beta-adrenergic receptor kinase, cAMP-dependent protein kinase, and protein kinase C occurs via distinct molecular mechanisms.
  51. (2003). Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects.
  52. (2008). Diabetes-induced upregulation of urotensin II and its receptor plays an important role in TGF-beta1-mediated renal fibrosis and dysfunction.
  53. (2005). Dicer is required for embryonic angiogenesis during mouse development.
  54. (2004). Differential effect of urotensin II on vascular tone in normal subjects and patients with chronic heart failure.
  55. (2005). Effect of exogenous urotensin-II on vascular tone in skin microcirculation of patients with essential hypertension.
  56. (2007). Effects of receptor density on Nociceptin/OrphaninFQ peptide receptor desensitisation: studies using the ecdysone inducible expression system.
  57. EH (2000a). Human urotensin-II, the most potent mammalian vasoconstrictor identified to date, as a therapeutic target for the management of cardiovascular disease.
  58. (2004). Elevated plasma human urotensin-II-like immunoreactivity in ischemic cardiomyopathy.
  59. (2003). Elevated plasma levels of human urotensin-II immunoreactivity in congestive heart failure.
  60. (2004). Emerging roles of urotensin-II in cardiovascular disease.
  61. (1953). Enzyme secretion and the incorporation of P32 into phospholipides of pancreas slices.
  62. (1992). Epidermal growth factor induces dose-dependent calcium oscillations in single fura-2-loaded hepatocytes.
  63. (2008). Everything you wanted to know about small RNA but were afraid to ask.
  64. (1999). Evidence for a role of Trp proteins in the oxidative stress-induced membrane conductances of porcine aortic endothelial cells.
  65. (2001). Evolving concepts in G protein-coupled receptor endocytosis: the role in receptor desensitization and signaling.
  66. (1999). Expression and functional characterization of the cardiac muscle ryanodine receptor Ca(2+) release channel in Chinese hamster ovary cells.
  67. (1962). Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan,
  68. (1999). Fluorescent calcium indicators based on BAPTA. In: Putney Jr, J (ed). Calcium Signaling.
  69. (2006). Fluorescent measurement of [Ca2+]c: basic pratical considerations. In: Lambert, D (ed). Calcium Signaling Protocols.
  70. (1998). G protein-coupled receptor adaptation mechanisms.
  71. (1996). G-protein-coupled receptor kinases.
  72. (2006). GRKs and beta-arrestins: roles in receptor silencing, trafficking and signaling.
  73. (1995). Heterologous desensitization of both phosphoinositide and Ca2+ signaling in SH-SY5Y neuroblastoma cells: a role for intracellular Ca2+ store depletion?
  74. (2000). Human urotensin II mediates vasoconstriction via an increase in inositol phosphates.
  75. (1999). Human urotensin-II is a potent vasoconstrictor and agonist for the orphan receptor GPR14.
  76. (2000). Human urotensin-II is an endothelium-dependent vasodilator in rat small arteries.
  77. (2004). Identification and pharmacological characterization of native, functional human urotensinII receptors in rhabdomyosarcoma cell lines.
  78. Identification of nonpeptidic urotensin II receptor antagonists by virtual screening based on a pharmacophore model derived from structure-activity relationships and nuclear magnetic resonance studies on urotensin II.
  79. (1999). Identification of the natural ligand of an orphan G-protein-coupled receptor involved in the regulation of vasoconstriction.
  80. (1999). Identification of urotensin II as the endogenous ligand for the orphan G-protein-coupled receptor GPR14.
  81. (2003). Identification of urotensin II-related peptide as the urotensin II-immunoreactive molecule in the rat brain.
  82. (2007). Increased human urotensin II levels are correlated with carotid atherosclerosis in essential hypertension.
  83. (2003). Increased plasma urotensin II levels in patients with diabetes mellitus.
  84. (1993). Increases in intracellular calcium via activation of an endogenous P2-purinoceptor in cultured CHO-K1 cells.
  85. (1990). Inositol phosphate formation and its relationship to calcium signaling.
  86. (1975). Inositol phospholipids and cell surface receptor function.
  87. (1993). Inositol trisphosphate and calcium signaling.
  88. (2003). Intracellular calcium handling in rat olfactory ensheathing cells and its role in axonal regeneration.
  89. (2005). Involvement of a cytoplasmic-tail serine cluster in urotensin II receptor internalization.
  90. (2008). Involvement of reactive oxygen species in urotensin II-induced proliferation of cardiac fibroblasts.
  91. (2006). Measurement of inositol(1,4,5)triphosphate using a stereospecific radioreceptor mass assay. In: Lambert,
  92. (2004). Mechanism of human urotensin II-induced contraction in rat aorta.
  93. (2003). Mechanisms regulating membrane trafficking of G protein-coupled receptors in the endocytic pathway.
  94. (2004). Messenger RNA turnover in eukaryotes: pathways and enzymes.
  95. (1990). Methods for receptor binding.
  96. (2008). MicroRNAs control gene expression: importance for cardiac development and pathophysiology.
  97. (2002). MicroRNAs: hidden in the genome.
  98. (2008). MicroRNAs: role in cardiovascular biology and disease.
  99. (2001). Modulation of human cytomegalovirus immediate-early gene enhancer by mitogen-activated protein kinase kinase kinase-1.
  100. (1985). Molecular mechanisms of receptor desensitization using the beta-adrenergic receptor-coupled adenylate cyclase system as a model.
  101. (1981). Mutations that lead to reiterations in the cell lineages of C.
  102. (2008). Neuropeptide interactions and REM sleep: a role for Urotensin II?
  103. (2008). New developments in the signaling mechanisms of the storeoperated calcium entry pathway.
  104. (2006). New roles for beta-arrestins in cell signaling: not just for seven-transmembrane receptors.
  105. (1989). NF-kappa B activation of the cytomegalovirus enhancer is mediated by a viral transactivator and by T cell stimulation.
  106. (2005). Nonpeptidic urotensin-II receptor antagonists I: in vitro pharmacological characterization of SB-706375.
  107. (2007). Nuclear factor-kappaB: activation and regulation during toll-like receptor signaling.
  108. (2008). Palosuran inhibits binding to primate UT receptors in cell membranes but demonstrates differential activity in intact cells and vascular tissues.
  109. (2006). Pharmacodynamics and pharmacokinetics of the urotensin II receptor antagonist palosuran in macroalbuminuric, diabetic patients.
  110. (2007). Pharmacological characterisation of the Urotensin-II receptor ligands urantide and UFP-803 in CHOhUT cells. 33° Congresso Nazionale della Societa’ Italiana di Farmacologia.
  111. (1996). Pharmacological principles for analysing responses of vascular smooth muscle. In: Garland CJ, A, JA (ed). Pharmacology of vascular smooth muscle. 1 edn.
  112. (1998). Phosphoinositide metabolism and Ca2+ oscillation.
  113. (2002). Plasma levels and cardiovascular gene expression of urotensin-II in human heart failure.
  114. (2002). Plasma urotensin II in heart failure.
  115. (2002). Plasma urotensin in human systolic heart failure.
  116. (1999). Properties of heterologously expressed hTRP3 channels in bovine pulmonary artery endothelial cells.
  117. (1951). Protein measurement with the Folin phenol reagent.
  118. (2006). Quantification of mRNA using real-time RTPCR.
  119. (1997). Radioligand binding methods for membrane preparations and intact cells. In: Challiss,
  120. (1982). Rapid and reversible disappearance of beta-adrenergic cell surface receptors.
  121. (2006). Ratiometric Ca2+ measurements using the Flexstation Scanning Fluorometer. In: Lambert, DG (ed). Calcium Signaling Protocols.
  122. (1996). Real time quantitative PCR.
  123. (2005). Real-time PCR for mRNA quantitation.
  124. (2007). Realtime PCR analysis of RNA extracted from formalin-fixed and paraffin-embeded tissues: effects of the fixation on outcome reliability.
  125. (1999). Receptor phosphorylation does not mediate cross talk between muscarinic M(3) and bradykinin B(2) receptors.
  126. (1998). Receptor-activated Ca2+ influx via human Trp3 stably expressed in human embryonic kidney (HEK)293 cells. Evidence for a noncapacitative Ca2+ entry.
  127. (1992). Regulation of cytosolic free calcium in isolated rat osteoclasts by calcitonin.
  128. (1999). Regulation of human cytomegalovirus gene expression.
  129. (1997). Regulation of Receptor Expression: Analysis of Receptor mRNA and Gene Transcription.
  130. (1973). Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction.
  131. (2006). Renal and vascular actions of urotensin II.
  132. (2006). Renal effects of human urotensin-II in rats with experimental congestive heart failure.
  133. RN (2000b). Differential vasoconstrictor activity of human urotensin-II in vascular tissue isolated from the rat, mouse, dog, pig, marmoset and cynomolgus monkey.
  134. (1998). Role of conformational constraints of position 7 of the disulphide bridge of h-alpha-CGRP derivatives in their agonist versus antagonist properties.
  135. (2007). Role of urotensin II and its receptor in health and disease.
  136. (2003). Role of urotensin II gene in genetic susceptibility to Type 2 diabetes mellitus in Japanese subjects.
  137. (2001). Role of urotensin II in patients on dialysis.
  138. (2004). Role of urotensin II in peripheral tissue as an autocrine/paracrine growth factor.
  139. (2007). Role of urotensin-II and its receptor in health and diseases.
  140. (1990). Selective inhibition of receptor-coupled phospholipase C-dependent processes in human platelets and polymorphonuclear neutrophils.
  141. (2007). Signaling to NF-kappaB by Toll-like receptors.
  142. (2006). Single cell and subcellular measurement of intracellular Ca2+ concentration In: Lamber, D (ed). Calcium signaling protocols. 2nd edn.
  143. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
  144. (2008). Statedependent calcium mobilization by urotensin-II in cultured human endothelial cells.
  145. (1997). Stimulus-dependent phosphorylation of G-protein-coupled receptors by casein kinase 1alpha.
  146. (2008). Structure activity relationship study on position 4 in the urotensin-II receptor ligand U-II(4-11). Peptides.
  147. (2005). Structure-activity relationship study on human urotensin II.
  148. (2004). Structure-activity relationships and structural conformation of a novel urotensin II-related peptide.
  149. Structure-function analysis of urotensin II and its use in the construction of a ligandreceptor working model.
  150. (2003). Structureactivity relationships of human urotensin II and related analogues on rat aortic ring contraction.
  151. (1990). Target amplification for DNA analysis by the polymerase chain reaction.
  152. (2000). Techniques to study the pharmacodynamics of isolated large and small blood vessels.
  153. (2000). The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans.
  154. (2002). The Basics of RT-PCR.
  155. (2007). The evasive nature of drug efficacy: implications for drug discovery.
  156. (2007). The G protein-coupled receptor kinase (GRK) interactome: role of GRKs in GPCR regulation and signaling.
  157. (2003). The neuromedin B receptor antagonist, BIM-23127, is a potent antagonist at human and rat urotensin-II receptors.
  158. (2006). The peptidic urotensin-II receptor ligand GSK248451 possesses less intrinsic activity than the low-efficacy partial agonists SB-710411 and urantide in native mammalian tissues and recombinant cell systems.
  159. (2006). The real-time polymerase chain reaction.
  160. (2004). The relationship between urotensin II plasma immunoreactivity and left ventricular filling pressures in coronary artery disease.
  161. (2002). The role of beta-arrestins in the termination and transduction of G-protein-coupled receptor signals.
  162. (2006). The role of urotensin II in cardiovascular and renal physiology and diseases.
  163. (2006). The urotensin-II receptor antagonist palosuran improves pancreatic and renal function in diabetic rats.
  164. (1996). trp, a novel mammalian gene family essential for agonist-activated capacitative Ca2+ entry.
  165. (2004). Urantide mimics urotensin-II induced calcium release in cells expressing recombinant UT receptors.
  166. (2003). Urantide: an ultrapotent urotensin II antagonist peptide in the rat aorta.
  167. (2005). Urotensin II in patients with chronic heart failure.
  168. (2007). Urotensin II is a new chemotactic factor for UT receptor-expressing monocytes.
  169. (2008). Urotensin II is an autocrine/paracrine growth factor for aortic adventitia of rat.
  170. (2007). Urotensin II is raised in acute myocardial infarction and low levels predict risk of adverse clinical outcome in humans.
  171. (1999). Urotensin II is the endogenous ligand of a G-protein-coupled orphan receptor, SENR (GPR14).
  172. (2004). Urotensin II, a novel peptide in central and peripheral cardiovascular control.
  173. (1980). Urotensin II: a somatostatin-like peptide in the caudal neurosecretory system of fishes.
  174. (2007). Urotensin II: from osmoregulation in fish to cardiovascular regulation in man.
  175. (2005). Urotensin II: its function in health and its role in disease.
  176. (1994). Voltage-dependent ionic conductances in Chinese hamster ovary cells.
  177. WG (2004a). Urotensin II: the old kid in town.
  178. WG (2004b). Urotensin II promotes hypertrophy of cardiac myocytes via mitogen-activated protein kinases.
  179. (1999). What molecular events underlie heterologous desensitization? Focus on "receptor phosphorylation does not mediate cross talk between muscarinic M(3) and bradykinin B(2) receptors".

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.