Differential regulation of cAMP-mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors

Eun Jin Lee, Soo Hyun Lee, Jin Won Jung, Weontae Lee, Byung Jin Kim, Kye Won Park, Sung Kil Lim, Chang Ju Yoon, Ja Hyun Baik

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)582-591
Number of pages10
JournalEuropean Journal of Biochemistry
Volume268
Issue number3
DOIs
Publication statusPublished - 2001 Sep 27

Fingerprint

Receptor, Melanocortin, Type 3
Melanocortin Receptors
Receptor, Melanocortin, Type 4
Transcription
Melanocyte-Stimulating Hormones
Genes
Ligands
Luciferases
GTP-Binding Proteins
Melanocortins
HEK293 Cells
Feeding Behavior
G-Protein-Coupled Receptors
Reporter Genes
Adenylyl Cyclases
Gene expression
Assays
Cells
Nuclear magnetic resonance
Hormones

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Lee, Eun Jin ; Lee, Soo Hyun ; Jung, Jin Won ; Lee, Weontae ; Kim, Byung Jin ; Park, Kye Won ; Lim, Sung Kil ; Yoon, Chang Ju ; Baik, Ja Hyun. / Differential regulation of cAMP-mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors. In: European Journal of Biochemistry. 2001 ; Vol. 268, No. 3. pp. 582-591.
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abstract = "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.",
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Differential regulation of cAMP-mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors. / Lee, Eun Jin; Lee, Soo Hyun; Jung, Jin Won; Lee, Weontae; Kim, Byung Jin; Park, Kye Won; Lim, Sung Kil; Yoon, Chang Ju; Baik, Ja Hyun.

In: European Journal of Biochemistry, Vol. 268, No. 3, 27.09.2001, p. 582-591.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Differential regulation of cAMP-mediated gene transcription and ligand selectivity by MC3R and MC4R melanocortin receptors

AU - Lee, Eun Jin

AU - Lee, Soo Hyun

AU - Jung, Jin Won

AU - Lee, Weontae

AU - Kim, Byung Jin

AU - Park, Kye Won

AU - Lim, Sung Kil

AU - Yoon, Chang Ju

AU - Baik, Ja Hyun

PY - 2001/9/27

Y1 - 2001/9/27

N2 - 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.

AB - 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.

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