Agonist-specific, high-affinity binding epitopes are contributed by an arginine in the N-terminus of the human oxytocin receptor
Abstract
The effects of the peptide hormone oxytocin (OT) are mediated by the oxytocin receptor, which is a member of the G-protein-coupled receptor family. Defining differences between the binding of agonists and antagonists to the OTR, at the molecular level, is of fundamental importance to understanding OTR activation and to rational drug design. Previous reports have indicated that the N-terminus of the OTR is required for OT binding. The aim of this study was to identify which individual residues within the N-terminal domain of the human OTR provided these OT binding epitopes. A series of truncated OTRs and mutant receptor constructs with systematic alanine substitution were characterized with respect to their pharmacological profile and intracellular signaling capability. Although a number of residues within the OTR will be required for optimal OT-OTR interaction, our data establish that Arg(34) within the N-terminal domain contributes to high-affinity OT binding. Removal of Arg(34) by truncation or substitution resulted in a 2000-fold decrease in OT affinity. In addition, we show that the arginyl at this locus is required for high-affinity binding of agonists in general. However, the importance of Arg(34) is restricted to agonist interaction with the OTR, as it was not required for binding peptide antagonist or non-peptide antagonist. It is noteworthy that the corresponding Arg in the related rat V(1a) vasopressin receptor is also required for high-affinity agonist binding. This study defines, at the molecular level, the role of the N-terminus of the OTR in high-affinity agonist binding and identifies a key residue for this function.
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Included in these substitutions are R33Q and N35D.
“Included in these substitutions are R33Q (a substitution also present in rats, voles, sheep, and cows; Wesley et al., 2002) and N35D.”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Wesley et al., 2002
R33 and N35 flank Arg34, a residue critical for OT binding.
“These residues flank Arg34, which is a residue critical for OT binding (Wesley et al., 2002)”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Wesley et al., 2002
Replacement of Arg33 and Asn35 with Ala does not affect OT binding.
“though replacement of Arg33 and Asn35 with Ala does not affect OT binding”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Wesley et al., 2002
The callitrichine N35D substitution is homologous to D33 in the vasopressin V2 receptor, adjacent to its critical R32.
“the callitrichine N35D substitution is homologous to the D33 in the vasopressin V2 receptor, which is adjacent to its critical R32 (Wesley et al., 2002).”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Wesley et al., 2002
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- Authors
- Wesley, V. J., Hawtin, S. R., Howard, H. C., & Wheatley, M.
- Affiliations
- University of Birmingham (GB)
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2002-03-27
- Volume
- 41
- Issue
- 16
- Pages
- 5086-5092
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2083831789
- Cited by
- 41
- References
- 12
- Citations by year
- 2026: 1 · 2025: 1 · 2023: 2 · 2022: 2 · 2021: 1 · 2020: 1 · 2019: 1 · 2018: 2 · 2017: 1 · 2016: 1 · 2015: 2 · 2014: 1 · 2013: 4 · 2012: 1
- FWCI
- 0.81
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- University of Birmingham Research Portal (University of Birmingham) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
- Keywords
- oxytocin receptor, N-terminal domain, G protein-coupled receptors, receptor truncation, alanine mutants, peptide antagonists, vasopressin V1a receptor, receptor activation
- MeSH terms
- Amino Acid Sequence, Epitopes, Arginine, Cell Line, Humans, Intracellular Fluid, Ligands, Molecular Sequence Data, Peptide Fragments, Protein Binding, Signal Transduction, Mutagenesis, Site-Directed, Receptors, Oxytocin
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