Characterisation of melanocortin receptor subtypes by radioligand binding analysis.
The DNAs encoding three melanocortin receptor subtypes (melanocortin MC1 receptor, melanocortin MC3 receptor and melanocortin MC5 receptor) were expressed individually in COS (CV-1 Origin, SV40) cells to characterise their ligand binding properties. The results indicated that [125I][Nle4, D-Phe7]alpha-MSH (melanocyte stimulating hormone) bound to a single saturable site with Kd values of 85.1 +/- 8.0 pmol/l (mean +/- S.E.M), 396 +/- 65 pmol/l and 5.05 +/- 1.00 nmol/l for melanocortin MC1 receptor, melanocortin MC3 receptor and melanocortin MC3 receptor, respectively. The melanocortin MC1 receptor and the melanocortin MC5 receptor showed a similar potency order to the melanocortic peptides examined which was markedly different from the potency order of the melanocortin MC3 receptor. The melanocortin MC1 receptor and melanocortin MC5 receptor had a relatively higher affinity for alpha-MSH than gamma-MSH and beta-MSH, whereas the melanocortin MC3 receptor had higher affinity for desacetyl-alpha-MSH, gamma-MSH and beta-MSH compared to alpha-MSH. The inclusion of the endopeptidase inhibitor phosphoramidon to prevent the breakdown of ACTH-(1-39) (adrenocorticotrophic hormone) to alpha-MSH, decreased ACTH-(1-39) binding affinity showing that ACTH-(1-39) had a much lower affinity for melanocortin MC1 receptor than reported earlier.
Schiöth
H B
HB
Department of Pharmaceutical Pharmacology, Biomedical Centre, Uppsala, Sweden.
Muceniece
R
R
Wikberg
J E
JE
Chhajlani
V
V
D000324:Adrenocorticotropic Hormone / Q000031:analogs & derivatives / Q000378:metabolism; D000595:Amino Acid Sequence; D001667:Binding, Competitive; D004305:Dose-Response Relationship, Drug; D006801:Humans; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D011869:Radioligand Assay; D018043:Receptors, Corticotropin / Q000145:classification* / Q000378:metabolism; D044101:Receptors, Melanocortin; D000521:alpha-MSH / Q000378:metabolism
D010446:Peptide Fragments; D018043:Receptors, Corticotropin; D044101:Receptors, Melanocortin; C013765:Org 2766; D000521:alpha-MSH; D000324:Adrenocorticotropic Hormone; D009074:Melanocyte-Stimulating Hormones