Abstract
Calcitonin gene-related peptide (CGRP) and adrenomedullin (ADM), two closely related peptides, initiate their biological responses through their interaction with calcitonin receptor-like receptor (CRLR). The CRLR receptor phenotype can be determined by coexpression of CRLR with one of the three-receptor activity modifying proteins (RAMPs). In this report, we characterized the pharmacological properties of the human or porcine CRLR with individual RAMPs transiently expressed in human embroynic kidney cell line (HEK-293). Characterization of RAMP1/human or porcine CRLR combination by radioligand binding ([125I] hαCGRP) and functional assay (activation of adenylyl cyclase) revealed the properties of CGRP receptor. Similarly characterization of RAMP2/human or porcine CRLR and RAMP3/human or porcine CRLR combination by radioligand binding ([125I]rADM) and functional assay (activation of adenylyl cyclase) revealed the properties of ADM (22–52) sensitive-ADM receptor. In addition, porcine CRLR/RAMP2 or 3 combination displayed specific high affinity [125I] hαCGRP binding also. Also, co-transfection of porcine CRLR with RAMPs provided higher expression level of the receptor than the human counterpart. Thus the present study along with earlier studies strongly support the role of RAMPs in the functional expression of specific CRLRs.
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References
Poyner DR: Pharmacology of receptors for calcitonin gene-related peptide and amylin. Trends Pharmacol Sci 16: 424–428, 1995
Wimalawansa SJ: Calcitonin gene-related peptide and its receptors: Molecular genetics, physiology, pathophysiology, and therapeutic potentials. Endocr Rev 17: 533–585, 1997
Samson WK: Proadrenomedullin-derived peptides. Front Neuroendocrinol 19: 100–127, 1998
Entzeroth M, Doods H, Wieland H, Wienen W: Adrenomedullin mediates vasodilation via CGRP1 receptors. Life Sci 56: PL 19–25, 1995
Nossaman BD, Feng CJ, Cheng DY, Dewitt BJ, Coy DH, Murphy WA, Kadowitz PJ: Comparative effects of adrenomedullin, an adrenomedullin analog, and CGRP in the pulmonary vascular bed of the cat and rat. Life Sci 56: PL63–PL66, 1995
Nuki C, Kawasaki H, Kitamura K, Takenaga M., Kanagawa K, Eto T, Wada A: Vasodilator effect of adrenomedullin and calcitonin generelated peptide receptors in rat mesenteric vascular beds. Biochem Biophys Res Commun 196: 245–251, 1993
Njuki F, Nicholl CG, Howard A, Mak JC, Barnes PJ, Girgis SI, Legon S: A new calcitonin-receptor-like sequence in rat pulmonary blood vessels. Clin Sci 85: 385–388, 1993
Fluhmann B, Muff R, Hunziker W, Fischer JA, Born W: A human orphan calcitonin receptor-like structure. Biochem Biophys Res Commun 206: 341–347, 1995
Aiyar N, Rand K, Elshourbagy NA, Zeng Z, Adamou JE, Bergsma DJ, Li Y: cDNA encoding the calcitonin gene-related peptide type 1 receptor. J Biol Chem 271: 11325–11329, 1996
Han ZQ, Coppock HA, Smith DM, Van Noorden S, Makgoba MW, Nicholl CG, Legon S: The interaction of CGRP and adrenomedullin with a receptor expressed in the pulmonary vascular endothelium. J Mol Endocrinol 18: 267–272, 1997
Elshourbagy NA, Adamou JE, Swift AM, Disa J, Mao J, Ganguly S, Bergsma DJ, Aiyar N: Molecular cloning and characterization of the porcine calcitonin gene-related peptide receptor. Endocrinology 139: 1678–1683, 1998
Quirion R, Van Rossum D, Dumont Y, St. Pierre S, Fournier A: Characterization of CGRP1 and CGRP2 receptor subtypes. Ann NY Acad Sci 657: 88–105, 1992
McLatchie LM, Fraser NJ, Main MJ, Wise A, Brown J, Thompson N, Solari R, Lee MG, Foord SM: RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor. Nature 393: 333–339, 1998
Christopoulos G, Perry KJ, Morfis M, Tilakatne N, Gao Y, Fraser NJ, Main MJ, Foord SM, Sexton PM: Multiple amylin receptors arise from receptor activity modifying protein interaction with the calcitonin receptor gene product. Mol Pharmacol 56: 235–242, 1999
Foord SM, Marshall FH: RAMPs: Accessory proteins for seven transmembrane domain receptors. Trends Pharmacol Sci 20: 184–187, 1999
Fraser NJ, Wise A, Brown J, McLatchie LM, Main MJ, Foord SM: The amino terminus of receptor activity modifying proteins is a critical determinant of glycosylation state and ligand binding of calcitonin-like receptor. Mol Pharmacol 55: 1054–1059, 1999
Tilakaratne N, Christopoulos G, Zumpe ET, Foord SM, Sexton PM: Amylin receptor phenotypes derived from human calcitonin receptor/ RAMP coexpression exhibit pharmacological differences dependent on receptor isoform and host cell environment. J Pharmacol Exp Ther 294: 61–72, 2000
Aiyar N, Baker E, Martin J, Patel A, Stadel JM, Willette RN, Barone FC: Differential calcitonin gene-related peptide (CGRP) and amylin binding sites in nucleus accumbens and lung: Potential models for studying CGRP/amylin receptor subtypes. J Neurochem 65: 1131–1138, 1995
Salmon Y, Londos C, Rodbell M: A highly selective adenylate cyclase assay. Anal Biochem 58: 541–548, 1974
Dang K, Disa J, Gout B, Aiyar N: Comparative affinities of adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) for [125I] AM and [125I] CGRP specific binding sites in porcine tissues. J Receptor Res 19: 803–817, 1999
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Aiyar, N., Disa, J., Pullen, M. et al. Receptor activity modifying proteins interaction with human and porcine calcitonin receptor-like receptor (CRLR) in HEK-293 cells. Mol Cell Biochem 224, 123–133 (2001). https://doi.org/10.1023/A:1011907328682
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DOI: https://doi.org/10.1023/A:1011907328682