Serine residues 110 and 114 are required for agonist binding but not antagonist binding to the melatonin MT1 receptor

S Conway, E S Mowat, Janice Drew, Perry Barrett, P Delagrange, Peter John Morgan

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Site-directed mutation of serine 110 (Ser(3.35)) and serine 114 (Ser(3.39)) in the human melatonin MT1, receptor to alanine residues reduced ligand binding affinities of seven known melatonin receptor agonists and partial agonists by 3- to 15-fold. These mutants also displayed a relative reduction in their affinities for melatonin-mediated functional responses of 30- and 14-fold, respectively. In contrast to the observed effects of the agonists and partial agonists, the melatonin receptor antagonist luzindole was found to bind to mutants Ser(3.35)Ala and Ser(3.39)Ala with affinities equivalent to that determined for the wild-type melatonin MT1 receptor. Luzindole was subsequently confirmed as an antagonist of melatonin-mediated functional responses for both mutant receptors. These studies have identified that in the human melatonin MT1 receptor, Ser(3.35) and Ser(3.39), in transmembrane domain 3, are critical for the formation of the high-affinity ligand binding site for agonists and partial agonists but not for the antagonist luzindole. (C) 2001 Academic Press.

Original languageEnglish
Pages (from-to)1229-1236
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume282
Issue number5
DOIs
Publication statusPublished - 20 Apr 2001

Keywords

  • melatonin
  • MT1 receptor
  • agonist
  • antagonist
  • ligand binding
  • transmembrane domain 3
  • G protein-coupled receptor
  • site-directed mutagenesis
  • beta-adrenergic-receptor
  • protein-coupled receptors
  • ovine pars tuberalis
  • ligand-binding
  • human MEL(1A)
  • cloning
  • identification
  • inhibition
  • subtypes
  • roles

Cite this

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title = "Serine residues 110 and 114 are required for agonist binding but not antagonist binding to the melatonin MT1 receptor",
abstract = "Site-directed mutation of serine 110 (Ser(3.35)) and serine 114 (Ser(3.39)) in the human melatonin MT1, receptor to alanine residues reduced ligand binding affinities of seven known melatonin receptor agonists and partial agonists by 3- to 15-fold. These mutants also displayed a relative reduction in their affinities for melatonin-mediated functional responses of 30- and 14-fold, respectively. In contrast to the observed effects of the agonists and partial agonists, the melatonin receptor antagonist luzindole was found to bind to mutants Ser(3.35)Ala and Ser(3.39)Ala with affinities equivalent to that determined for the wild-type melatonin MT1 receptor. Luzindole was subsequently confirmed as an antagonist of melatonin-mediated functional responses for both mutant receptors. These studies have identified that in the human melatonin MT1 receptor, Ser(3.35) and Ser(3.39), in transmembrane domain 3, are critical for the formation of the high-affinity ligand binding site for agonists and partial agonists but not for the antagonist luzindole. (C) 2001 Academic Press.",
keywords = "melatonin, MT1 receptor, agonist, antagonist, ligand binding, transmembrane domain 3, G protein-coupled receptor, site-directed mutagenesis, beta-adrenergic-receptor, protein-coupled receptors, ovine pars tuberalis, ligand-binding, human MEL(1A), cloning, identification, inhibition, subtypes, roles",
author = "S Conway and Mowat, {E S} and Janice Drew and Perry Barrett and P Delagrange and Morgan, {Peter John}",
year = "2001",
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language = "English",
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journal = "Biochemical and Biophysical Research Communications",
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TY - JOUR

T1 - Serine residues 110 and 114 are required for agonist binding but not antagonist binding to the melatonin MT1 receptor

AU - Conway, S

AU - Mowat, E S

AU - Drew, Janice

AU - Barrett, Perry

AU - Delagrange, P

AU - Morgan, Peter John

PY - 2001/4/20

Y1 - 2001/4/20

N2 - Site-directed mutation of serine 110 (Ser(3.35)) and serine 114 (Ser(3.39)) in the human melatonin MT1, receptor to alanine residues reduced ligand binding affinities of seven known melatonin receptor agonists and partial agonists by 3- to 15-fold. These mutants also displayed a relative reduction in their affinities for melatonin-mediated functional responses of 30- and 14-fold, respectively. In contrast to the observed effects of the agonists and partial agonists, the melatonin receptor antagonist luzindole was found to bind to mutants Ser(3.35)Ala and Ser(3.39)Ala with affinities equivalent to that determined for the wild-type melatonin MT1 receptor. Luzindole was subsequently confirmed as an antagonist of melatonin-mediated functional responses for both mutant receptors. These studies have identified that in the human melatonin MT1 receptor, Ser(3.35) and Ser(3.39), in transmembrane domain 3, are critical for the formation of the high-affinity ligand binding site for agonists and partial agonists but not for the antagonist luzindole. (C) 2001 Academic Press.

AB - Site-directed mutation of serine 110 (Ser(3.35)) and serine 114 (Ser(3.39)) in the human melatonin MT1, receptor to alanine residues reduced ligand binding affinities of seven known melatonin receptor agonists and partial agonists by 3- to 15-fold. These mutants also displayed a relative reduction in their affinities for melatonin-mediated functional responses of 30- and 14-fold, respectively. In contrast to the observed effects of the agonists and partial agonists, the melatonin receptor antagonist luzindole was found to bind to mutants Ser(3.35)Ala and Ser(3.39)Ala with affinities equivalent to that determined for the wild-type melatonin MT1 receptor. Luzindole was subsequently confirmed as an antagonist of melatonin-mediated functional responses for both mutant receptors. These studies have identified that in the human melatonin MT1 receptor, Ser(3.35) and Ser(3.39), in transmembrane domain 3, are critical for the formation of the high-affinity ligand binding site for agonists and partial agonists but not for the antagonist luzindole. (C) 2001 Academic Press.

KW - melatonin

KW - MT1 receptor

KW - agonist

KW - antagonist

KW - ligand binding

KW - transmembrane domain 3

KW - G protein-coupled receptor

KW - site-directed mutagenesis

KW - beta-adrenergic-receptor

KW - protein-coupled receptors

KW - ovine pars tuberalis

KW - ligand-binding

KW - human MEL(1A)

KW - cloning

KW - identification

KW - inhibition

KW - subtypes

KW - roles

U2 - 10.1006/bbrc.2001.4722

DO - 10.1006/bbrc.2001.4722

M3 - Article

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EP - 1236

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

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