Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor

Denise L. Wootten, John Simms, Amelia J. Massoura, Julie E. Trim, Mark Wheatley*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

Defining key differences between agonist and antagonist binding to hormone receptors is important and will aid rational drug design. Glu1.35 in transmembrane helix 1 (TM1) of the human oxytocin receptor (OTR) is absolutely conserved in all OTRs cloned to date. We establish that Glu1.35 is critical for high affinity binding of agonists (full and partial) but is not required for antagonist binding (peptide or non-peptide). Consequently, the mutant receptor [E1.35A]OTR exhibited markedly decreased OT affinity (>1200-fold) and disrupted second messenger generation. Substitutions of Glu1.35 by Asp, Gln or Arg were incapable of supporting wild-type OTR agonist binding or signaling. Molecular modeling revealed that Glu1.35 projects into the receptor's central binding crevice and provides agonist-specific contacts not utilized by antagonists. This study explains why Glu is absolutely conserved at residue-1.35 in all receptors binding OT and related peptides, and provides molecular insight into key differences between agonist-receptor and antagonist-receptor binding modes.

Original languageEnglish
Pages (from-to)20-27
Number of pages8
JournalMolecular and Cellular Endocrinology
Volume333
Issue number1
DOIs
Publication statusPublished - 10 Feb 2011

Fingerprint

Oxytocin Receptors
Glutamic Acid
Peptides
Molecular modeling
Drug Design
Second Messenger Systems
Viperidae
Substitution reactions
Hormones
Pharmaceutical Preparations
human OXTR protein

Keywords

  • GPCR
  • Oxytocin
  • Oxytocin receptor
  • Peptide hormone

Cite this

Wootten, D. L., Simms, J., Massoura, A. J., Trim, J. E., & Wheatley, M. (2011). Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor. Molecular and Cellular Endocrinology, 333(1), 20-27. https://doi.org/10.1016/j.mce.2010.11.029
Wootten, Denise L. ; Simms, John ; Massoura, Amelia J. ; Trim, Julie E. ; Wheatley, Mark. / Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor. In: Molecular and Cellular Endocrinology. 2011 ; Vol. 333, No. 1. pp. 20-27.
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Wootten, DL, Simms, J, Massoura, AJ, Trim, JE & Wheatley, M 2011, 'Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor', Molecular and Cellular Endocrinology, vol. 333, no. 1, pp. 20-27. https://doi.org/10.1016/j.mce.2010.11.029

Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor. / Wootten, Denise L.; Simms, John; Massoura, Amelia J.; Trim, Julie E.; Wheatley, Mark.

In: Molecular and Cellular Endocrinology, Vol. 333, No. 1, 10.02.2011, p. 20-27.

Research output: Contribution to journalArticle

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AU - Simms, John

AU - Massoura, Amelia J.

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AU - Wheatley, Mark

PY - 2011/2/10

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N2 - Defining key differences between agonist and antagonist binding to hormone receptors is important and will aid rational drug design. Glu1.35 in transmembrane helix 1 (TM1) of the human oxytocin receptor (OTR) is absolutely conserved in all OTRs cloned to date. We establish that Glu1.35 is critical for high affinity binding of agonists (full and partial) but is not required for antagonist binding (peptide or non-peptide). Consequently, the mutant receptor [E1.35A]OTR exhibited markedly decreased OT affinity (>1200-fold) and disrupted second messenger generation. Substitutions of Glu1.35 by Asp, Gln or Arg were incapable of supporting wild-type OTR agonist binding or signaling. Molecular modeling revealed that Glu1.35 projects into the receptor's central binding crevice and provides agonist-specific contacts not utilized by antagonists. This study explains why Glu is absolutely conserved at residue-1.35 in all receptors binding OT and related peptides, and provides molecular insight into key differences between agonist-receptor and antagonist-receptor binding modes.

AB - Defining key differences between agonist and antagonist binding to hormone receptors is important and will aid rational drug design. Glu1.35 in transmembrane helix 1 (TM1) of the human oxytocin receptor (OTR) is absolutely conserved in all OTRs cloned to date. We establish that Glu1.35 is critical for high affinity binding of agonists (full and partial) but is not required for antagonist binding (peptide or non-peptide). Consequently, the mutant receptor [E1.35A]OTR exhibited markedly decreased OT affinity (>1200-fold) and disrupted second messenger generation. Substitutions of Glu1.35 by Asp, Gln or Arg were incapable of supporting wild-type OTR agonist binding or signaling. Molecular modeling revealed that Glu1.35 projects into the receptor's central binding crevice and provides agonist-specific contacts not utilized by antagonists. This study explains why Glu is absolutely conserved at residue-1.35 in all receptors binding OT and related peptides, and provides molecular insight into key differences between agonist-receptor and antagonist-receptor binding modes.

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Wootten DL, Simms J, Massoura AJ, Trim JE, Wheatley M. Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor. Molecular and Cellular Endocrinology. 2011 Feb 10;333(1):20-27. https://doi.org/10.1016/j.mce.2010.11.029