Interaction of melanin-concentrating hormone (MCH), neuropeptide E-I (NEI), neuropeptide G-E (NGE), and alpha-MSH with melanocortin and MCH receptors on mouse B16 melanoma cells.
Melanin-concentrating hormone (MCH) and alpha-melanocyte-stimulating hormone (alpha-MSH) are known to exhibit mostly functionally antagonistic, but in some cases agonistic activities, e.g., in pigment cells and in the brain. Neuropeptide E-I (NEI) displays functional MCH-antagonist and MSH-agonist activity in different behavioral paradigms; the role of neuropeptide G-E (NGE) is not known. This study addressed the question of possible molecular interactions between alpha-MSH, MCH and the MCH-precursor-derived peptides NEI and NGE at the level of the pigment cell MCH receptor subtype (MCH-Rpc) and the different melanocortin (MC) receptors. Radioreceptor assays using [125I]MCH, [125l]alpha-MSH and [125I]NEI as radioligands and bioassays were performed with MCI-R-positive and MC1-R-negative mouse B16 melanoma cells and with COS cells expressing the different MC receptors. The IC50s of alpha-MSH and NEI or NGE for [125I]MCH displacement from mouse MCH-Rpc were 80-fold and, respectively, >300-fold higher than that of MCH, and the IC50s for MCH and NEI or NGE for [125I]alpha-MSH displacement from mouse MC1-R were 50,000-fold and >200,000-fold higher than that of alpha-MSH. No high-affinity binding sites for NEI were detected on B16 melanoma cells and there was no significant displacement of [1251]alpha-MSH by MCH, NEI or NGE with MC3-R, MC4-R and MC5-R expressed in COS cells. At concentrations of 100 nM to 10 microM, however, MCH, NEI and NGE induced cAMP formation and melanin synthesis which could be blocked by agouti protein or inhibitors of adenylate cyclase or protein kinase A. This shows that mammalian MCH-precursor-derived peptides may mimic MSH signalling via MC1-R activation at relatively high, but physiologically still relevant concentrations, as e.g. found in autocrine/paracrine signalling mechanisms.
Hintermann
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E
Laboratory of Endocrinology, Department of Research (ZLF), University Hospital and University Children's Hospital, CH-4031 Basel, Switzerland.
Tanner
H
H
Talke-Messerer
C
C
Schlumberger
S
S
Zumsteg
U
U
Eberle
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AN
D000818:Animals; D001667:Binding, Competitive; D019556:COS Cells; D015870:Gene Expression; D006801:Humans; D007028:Hypothalamic Hormones / Q000378:metabolism*; D007700:Kinetics; D008543:Melanins / Q000096:biosynthesis / Q000378:metabolism*; D008546:Melanoma, Experimental / Q000378:metabolism; D051379:Mice; D009842:Oligopeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010907:Pituitary Hormones / Q000378:metabolism*; D011869:Radioligand Assay; D044104:Receptor, Melanocortin, Type 3; D018043:Receptors, Corticotropin / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D044101:Receptors, Melanocortin; D011979:Receptors, Pituitary Hormone / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D000521:alpha-MSH / Q000378:metabolism*
D007028:Hypothalamic Hormones; C470053:Mc3r protein, mouse; D008543:Melanins; D009842:Oligopeptides; D010446:Peptide Fragments; D010907:Pituitary Hormones; D044104:Receptor, Melanocortin, Type 3; D018043:Receptors, Corticotropin; D044101:Receptors, Melanocortin; D011979:Receptors, Pituitary Hormone; D011994:Recombinant Proteins; C075863:melanin concentrating hormone precursor (109-129)-glycyl-glutamic acid; C092274:melanin-concentrating hormone receptor; C522850:neuropeptide EI; D000521:alpha-MSH; C019463:melanin-concentrating hormone