Melanocortin‐3 receptor responseis influenced by the melanocortin receptoraccessory protein‐1

Berruien, N., Le Tissier, P., Murray, J.F. and Smith, C.L. 2018. Melanocortin‐3 receptor responseis influenced by the melanocortin receptoraccessory protein‐1. Pharmacology 2018. Queen Elizabeth II conference centre London 18 - 20 Dec 2018 Wiley-Blackwell. https://doi.org/10.1111/bph.14681

TitleMelanocortin‐3 receptor responseis influenced by the melanocortin receptoraccessory protein‐1
AuthorsBerruien, N., Le Tissier, P., Murray, J.F. and Smith, C.L.
Abstract

Background and Purpose: Melanocortin receptors (MC1–5) areinvolved in controlling appetite, glucocorticoid production, and modu-lating anti‐inflammatory responses. These functions are mediatedthrough binding with different melanocortin peptides. MC2expressionand activation was previously found to be regulated by twomelanocortin receptor accessory proteins, MRAP1 and MRAP2. ThePRESTO‐TANGO (parallel receptorome expression and screening viatranscriptional output, with transcriptional activation followingarrestin translocation) assay was designed to study in vitro G‐proteincoupled signalling. This assay reports MC receptor activation bylinking beta‐arrestin recruitment to the transcription of luciferasewhich generates light from catalysis of its substrate, luciferin. Theaim of this study was to examine the human MC3responsiveness tothe melanocortin ligands when co‐expressed withMRAP1using the PRESTO‐TANGO in vitro assay.
Experimental Approach:Modified HEK293 (HTLA) cells weretransfected using PolyJet™with 5 ng of MC3‐Tango plasmid withand without co‐transfected 5‐ng MRAP1‐pCDNA3.1‐FLAG. TotalDNA per well of a 96 plate was 50 ng (supplemented by pCDNA3.1).The transfected cells were stimulated 48‐hr post‐transfection witheither ACTH(1–39), NDP‐α‐MSH, or D‐Trp8‐γ3‐MSH (1 × 10−6–1×10−12M) for 20 hr. Luminescence was determined using a lumi-nescent plate reader (GloMax®) and log dose–response curves gener-ated withn=4–6.MC3‐Tango stimulation with and without MRAP1co‐transfection was compared using unpaired two‐tailedttest.Key Results:EC50values determined with the PRESTO‐Tango assayfor MC3were ACTH(1–39), 51.24 nM; NDP‐α‐MSH, 153.3 nM; andD‐Trp8‐γ3‐MSH, 78.32 nM. The co‐expression ofMC3withMRAP1led to a significant increase in the sensitivity of MC3to ACTH(1–39)(35.43 nM,P= .0099,n= 6); however, the effect of co‐expressingMC3and MRAP1 with NDP‐α‐MSH (204 nM,P= .5122,n= 4) andTrp8‐γ3‐MSH (77.6 nM,P= .4964,n= 4) was not significant.
TABLE 1ReceptorMRAP1co‐expression LigandEC50± SEM (nM)nMC3—ACTH(1–39)51.24 ± 2.056MC3YesACTH(1–39)35.43 ± 3.456MC3—NDP‐α;;‐MSH 153.3 ± 10.144MC3YesNDP‐α;;‐MSH 204 ± 77.984MC3—D‐Trp8‐γ3‐MSH 78.32 ± 1.724MC3YesD‐Trp8‐γ3‐MSH 77.6 ± 1.654Conclusions and Implications:This is the first report about the study-ing co‐expression of human MC3with MRAP1 in an in vitro assay.MRAP1 significantly increases the MC3affinity to ACTH(1–39)butnot to NDP‐α;;‐MSH or D‐Trp8‐γ3‐MSH, showing that melanocortinreceptor accessory proteins specifically alter melanocortin receptoractivation by different melanocortin ligands.

Year2018
ConferencePharmacology 2018
PublisherWiley-Blackwell
Publication dates
Published21 May 2019
Published online09 Jul 2019
Published in printAug 2019
JournalBritish Journal of Pharmacology
Journal citation176 (16), p. 3052
ISSN0007-1188
Digital Object Identifier (DOI)https://doi.org/10.1111/bph.14681
Web address (URL) of conference proceedingshttps://bpspubs.onlinelibrary.wiley.com/doi/epdf/10.1111/bph.14681
Web address (URL)https://bpspubs.onlinelibrary.wiley.com/doi/epdf/10.1111/bph.14681

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Lea, R.G., Hannah, L.T., Redmer, D.A., Aitken, R.P., Milne, J.S., Fowler, P.A., Murray, J.F. and Wallace, J.M. 2005. Developmental indices of nutritionally induced placental growth restriction in the adolescent sheep. Pediatric Research. 57 (4), pp. 599-604. https://doi.org/10.1203/01.PDR.0000155949.08547.66

Pathophysiological concentrations of ADMA alter human coronary artery endothelial cell gene expression: an insight into the pathophysiological significance of raised plasma ADMA levels
Smith, C.L., Anthony, S., Malaki, M., Hubank, M., Leiper, J.M. and Vallance, P. 2005. Pathophysiological concentrations of ADMA alter human coronary artery endothelial cell gene expression: an insight into the pathophysiological significance of raised plasma ADMA levels. in: Boger, R., Fleming, I. and Waltenberger, J. (ed.) Vascular Biology and Medicine: 3rd European Meeting, Hamburg, September 2005, abstracts Germany Karger.

Potential therapeutic benefit of novel DDAH inhibitors for the treatment of endotoxemia
Nandi, M., Rossiter, S., Torondel, B., Malaki, M., Smith, C.L., Stidwill, R., Leiper, J.M. and Vallance, P. 2005. Potential therapeutic benefit of novel DDAH inhibitors for the treatment of endotoxemia. in: Boger, R., Fleming, I. and Waltenberger, J. (ed.) Vascular Biology and Medicine: 3rd European Meeting, Hamburg, September 2005, abstracts Germany Karger.

Evidence of systemic and pulmonary endothelial dysfunction in the Dimethylarginine Dimethylaminohydrolase I (DDAH I+/-) heterozygous knockout mouse
Malaki, M., Nandi, M., Madhani, M., Gill, H., Smith, C.L., Leiper, J.M. and Vallance, P. 2005. Evidence of systemic and pulmonary endothelial dysfunction in the Dimethylarginine Dimethylaminohydrolase I (DDAH I+/-) heterozygous knockout mouse. in: Boger, R., Fleming, I. and Waltenberger, J. (ed.) Vascular Biology and Medicine: 3rd European Meeting, Hamburg, September 2005, abstracts Germany Karger.

Dimethylarginine dimethylaminohydrolase I (DDAH I) heterozygous knockout mice display a cardiovascular phenotype consisting of pulmonary and systemic endothelial dysfunction
Malaki, M., Nandi, M., Madhani, M., Gill, H., Smith, C.L., Leiper, J.M. and Vallance, P. 2005. Dimethylarginine dimethylaminohydrolase I (DDAH I) heterozygous knockout mice display a cardiovascular phenotype consisting of pulmonary and systemic endothelial dysfunction. British Pharmaceutical Society Summer Meeting. Cambridge, UK 06-08 Jul 2005

Leptin expression in the fetus and placenta during mouse pregnancy
Malik, N.M., Carter, N.D., Wilson, C.A., Scaramuzzi, R.J., Stock, M.J. and Murray, J.F. 2005. Leptin expression in the fetus and placenta during mouse pregnancy. Placenta. 26 (1), pp. 47-52. https://doi.org/10.1016/j.placenta.2004.03.009

Cardiovascular tests: use & limits of biochemical markers - therapeutic measurements of ADMA involved in cardiovascular disorders
Smith, C.L. and Vallance, P. 2005. Cardiovascular tests: use & limits of biochemical markers - therapeutic measurements of ADMA involved in cardiovascular disorders. Current Pharmaceutical Design. 11 (17), pp. 2177-2185. https://doi.org/10.2174/1381612054367364

Selective substrate-based inhibitors of mammalian dimethylarginine dimethylaminohydrolase
Rossiter, S., Smith, C.L., Malaki, M., Nandi, M., Gill, H., Leiper, J.M., Vallance, P. and Selwood, D.L. 2005. Selective substrate-based inhibitors of mammalian dimethylarginine dimethylaminohydrolase. Journal of Medicinal Chemistry. 48 (14), pp. 1670-1678. https://doi.org/10.1021/jm050187a

Effects of ADMA upon gene expression: an insight into the pathophysiological significance of raised plasma ADMA
Smith, C.L., Anthony, S., Hubank, M., Leiper, J.M. and Vallance, P. 2005. Effects of ADMA upon gene expression: an insight into the pathophysiological significance of raised plasma ADMA. PLoS Medicine. 2 (10), pp. 1031-1043. https://doi.org/10.1371/journal.pmed.0020264

Effects of low dose ADMA on gene expression in human coronary artery endothelial cells
Smith, C.L., Leiper, J.M. and Vallance, P. 2004. Effects of low dose ADMA on gene expression in human coronary artery endothelial cells. Nitric Oxide: Biology and Chemistry. 11 (1), p. 59. https://doi.org/10.1016/j.niox.2004.07.003

Restoration of nitric oxide production by N-hydroxy-L-arginine in endothelial cells with NO deficiency triggered by elevated glucose
Crabtree, M.J., Smith, C.L. and Gross, S.S. 2004. Restoration of nitric oxide production by N-hydroxy-L-arginine in endothelial cells with NO deficiency triggered by elevated glucose. Nitric Oxide: Biology and Chemistry. 11 (1), p. 65. https://doi.org/10.1016/j.niox.2004.07.003

Overexpression of dimethylarginine dimethylaminohydrolase enhances tumour hypoxia: an insight into the relationship of hypoxia and angiogenesis In vivo
Kostourou, V., Troy, H., Murray, J.F., Cullis, E.R., Whitley, G.S.J., Griffiths, J.R. and Robinson, S. 2004. Overexpression of dimethylarginine dimethylaminohydrolase enhances tumour hypoxia: an insight into the relationship of hypoxia and angiogenesis In vivo. NeoPlasia. 6 (4), pp. 401-411. https://doi.org/10.1593/neo.04109

Effects of low dose ADMA on gene expression in human coronary artery endothelial cells
Smith, C.L., Leiper, J.M. and Vallance, P. 2004. Effects of low dose ADMA on gene expression in human coronary artery endothelial cells. 3rd International Conference on the Biology, Chemistry and Therapeutic Applications of Nitric Oxide: 4th Anuual Scientific Meeting of the Nitric Oxide Society of Japan. Nara, Japan 24-28 May 2004

Restoration of nitric oxide production by N-hydroxyarginine in endothelial cells with NO deficiency triggered by elevated glucose
Crabtree, M.J., Smith, C.L. and Gross, S.S. 2004. Restoration of nitric oxide production by N-hydroxyarginine in endothelial cells with NO deficiency triggered by elevated glucose. 3rd International Conference on the Biology, Chemistry and Therapeutic Applications of Nitric Oxide: 4th Anuual Scientific Meeting of the Nitric Oxide Society of Japan. Nara, Japan 24-28 May 2004

Neonatal 5HT activity antagonizes the masculinizing effect of testosterone on the luteinizing hormone release response to gonadal steroids and on brain structures in rats
Murray, J.F., Dakin, C.L., Siddiqui, A., Pellatt, L.J., Ahmed, S., Ormerod, L.J.A., Swan, A.V., Davies, D.C. and Wilson, C.A. 2004. Neonatal 5HT activity antagonizes the masculinizing effect of testosterone on the luteinizing hormone release response to gonadal steroids and on brain structures in rats. European Journal of Neuroscience. 19 (2), pp. 387-395. https://doi.org/10.1111/j.0953-816X.2003.03158.x

Central orexin A has site-specific effects on luteinizing hormone release in female rats
Small, C.J., Goubillon, M.L., Murray, J.F., Siddiqui, A., Grimshaw, S.E., Young, H., Sivanesan, V., Kalamatianos, T., Kennedy, A.R., Coen, C.W., Bloom, S.R. and Wilson, C.A. 2003. Central orexin A has site-specific effects on luteinizing hormone release in female rats. Endocrinology. 144 (7), pp. 3225-3236. https://doi.org/10.1210/en.2002-0041

Measurement of cardiac troponin I in striated muscle using three experimental methods
Fredericks, S., Bainbridge, K., Murray, J.F., Collinson, P.O., Carter, N.D. and Holt, D.W. 2003. Measurement of cardiac troponin I in striated muscle using three experimental methods. Annals of Clinical Biochemistry. 40 (3), pp. 244-248. https://doi.org/10.1258/000456303321610547

Dimethylarginine dimethylaminohydrolase activity modulates ADMA levels, VEGF expression, and cell phenotype
Smith, C.L., Birdsey, G.M., Anthony, S., Arrigoni, F.I., Leiper, J.M. and Vallance, P. 2003. Dimethylarginine dimethylaminohydrolase activity modulates ADMA levels, VEGF expression, and cell phenotype. Biochemical and Biophysical Research Communications. 308 (4), pp. 984-989. https://doi.org/10.1016/S0006-291X(03)01507-9

Cardiac troponin T and creatine kinase MB content in skeletal muscle of the uremic rat
Fredericks, S., Murray, J.F., Carter, N.D., Chesser, A.M.S., Papachristou, S., Yaqoob, M.M., Collinson, P.O., Gaze, D. and Holt, D.W. 2002. Cardiac troponin T and creatine kinase MB content in skeletal muscle of the uremic rat. Clinical Chemistry. 48 (6), pp. 859-868.

The redox status of bound pterin cofactor determines whether eNOS produces NO or superoxide anion: [3H] - BH4 binding studies provide insights into vascular pathophysiology
Jones, C.L., Vasquez-Vivar, J., Kalyanaraman, B., Griscavage-Ennis, J.M., Gross, S.S. and Smith, C.L. 2002. The redox status of bound pterin cofactor determines whether eNOS produces NO or superoxide anion: [3H] - BH4 binding studies provide insights into vascular pathophysiology. in: Milstien, S., Kapatos, G., Levine, R.A. and Shane, B. (ed.) Chemistry and biology of pteridines and folates: proceedings of the 12th International Symposium on Pteridines and Folates, National Institutes of Health, Bethesda, M.D. Boston, USA Kluwer Academic Publishers. pp. 271-276

Activation and inactivation of neuronal nitric oxide synthase: characterization of Ca2+-dependent [125I]Calmodulin binding
Weissman, B.A., Jones, C.L., Liu, Q., Gross, S.S. and Smith, C.L. 2002. Activation and inactivation of neuronal nitric oxide synthase: characterization of Ca2+-dependent [125I]Calmodulin binding. European Journal of Pharmacology. 435 (1), pp. 9-18. https://doi.org/10.1016/S0014-2999(01)01560-6

Leptin requirement for conception, implantation, and gestation in the mouse
Malik, N.M., Carter, N.D., Murray, J.F., Scaramuzzi, R.J., Wilson, C.A. and Stock, M.J. 2001. Leptin requirement for conception, implantation, and gestation in the mouse. Endocrinology. 142 (12), pp. 5198-5202.

Cardiac troponin T and creatine kinase MB are not increased in exterior oblique muscle of patients with renal failure
Fredericks, S., Murray, J.F., Bewick, M., Chang, R., Collinson, P.O., Carter, N.D. and Holt, D.W. 2001. Cardiac troponin T and creatine kinase MB are not increased in exterior oblique muscle of patients with renal failure. Clinical Chemistry. 47 (6), pp. 1023-1030.

Chapter 10: Tetrahydrobiopterin: An Essential Cofactor of Nitric Oxide Synthase with an Elusive Role
Smith, C.L. 2000. Chapter 10: Tetrahydrobiopterin: An Essential Cofactor of Nitric Oxide Synthase with an Elusive Role. in: Ignarro, L.J. (ed.) Nitric Oxide: Biology and Pathobiology Elsevier. pp. 167-185

Carbon monoxide induces vasodilation and nitric oxide release but suppresses endothelial NOS
Thorup, C., Jones, C.L., Gross, S.S., Moore, L.C. and Goligorsky, M.S. 1999. Carbon monoxide induces vasodilation and nitric oxide release but suppresses endothelial NOS. American journal of physiology. 277 (6), pp. F882-F889.

An autoinhibitory control element defines calcium-regulated isoforms of nitric oxide synthase
Salerno, J.C., Harris, D.E., Irizarry, K., Patel, B., Morales, A.J., Smith, S.M.E., Martasek, P., Roman, L.J., Masters, B.S.S., Jones, C.L., Weissman, B.A., Lane, P., Liu, Q., Gross, S.S. and Smith, C.L. 1997. An autoinhibitory control element defines calcium-regulated isoforms of nitric oxide synthase. Journal of Biological Chemistry. 272 (47), pp. 29769-29777.

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