Open AccessDifferential regulation of cAMP‐mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors
Author(s) -
Lee Eun Jin,
Lee SooHyun,
Jung JinWon,
Lee Weontae,
Kim Byung Jin,
Park Kye Won,
Lim SungKil,
Yoon ChangJu,
Baik JaHyun
Publication year - 2001
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.2001.01900.x
Subject(s) - melanocortin 3 receptor , melanocortin , hek 293 cells , receptor , microbiology and biotechnology , g protein coupled receptor , reporter gene , chemistry , melanocortin receptor , biology , gene , gene expression , biochemistry
Melanocortins are known to be involved in the regulation of feeding behavior. These hormones mediate their effects through G‐protein‐coupled receptors by stimulating adenylate cyclase. In this study we describe the functional response of melanocortin 4 receptor (MC4R) and melanocortin 3 receptor (MC3R) in HEK 293T cells, by using a luciferase reporter gene under the transcriptional control of a cAMP‐responsive element (CRE) as a monitor of intracellular cAMP levels and cAMP‐regulated gene expression. We were able to show that MC4R and MC3R expressed in the human cell line HEK 293T stimulate transcription induced by stimulation with different analogs of α‐melanocyte‐stimulating hormone (α‐MSH) at different levels. In our assay of CRE‐mediated gene transcription activity, α‐MSH‐ND was the most efficient α‐MSH analog for MC4R whereas NDP‐MSH was the most efficient for MC3R. Changing the His6 residue of α‐MSH‐ND to Gln or Lys markedly decreased CRE‐mediated luciferase activity for MC3R compared with MC4R. On analysis by modeling the receptor–ligand complex by NMR, [Gln6]α‐MSH‐ND and [Lys6]α‐MSH‐ND showed different conformational interactions between MC3R and MC4R. Furthermore, the maximum coupling efficiency of MC4R and MC3R to G proteins was different; MC4R showed only 30–50% of the maximum activity induced by MC3R. In total, our results suggest that a differential receptor–ligand interaction is involved and that the relative interactions of MC3R and MC4R with G protein are possibly quantitatively and qualitatively different.