The N-terminal juxtamembrane segment of the V1a vasopressin receptor provides two independent epitopes required for high-affinity agonist binding and signaling
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It is fundamentally important to define how agonist-receptor interaction differs from antagonist-receptor interaction. The V1a vasopressin receptor (V1aR) is a member of the neurohypophysial hormone subfamily of G protein-coupled receptors. Using alanine-scanning mutagenesis of the N-terminal juxtamembrane segment of the V1aR, we now establish that Glu54 (1.35) is critical for arginine vasopressin binding. The mutant [E54A]V1aR exhibited decreased arginine vasopressin affinity (1700-fold) and disrupted signaling, but antagonist binding was unaffected. Mutation of Glu54 had an almost identical pharmacological effect as mutation of Arg46, raising the possibility that agonist binding required a mutual interaction between Glu54 and Arg46. The role of these two charged residues was investigated by 1) substituting Glu54; 2) inserting additional Glu/Arg in transmembrane helix (TM) 1; 3) repositioning the Glu/Arg in TM1; and 4) characterizing the reciprocal mutant [R46E/E54R]V1aR. We conclude that 1) the positive/negative charges need to be precisely positioned in this N terminus/TM1 segment; and 2) Glu54 and Arg46 function independently, providing two discrete epitopes required for high-affinity agonist binding and signaling. This study explains why Glu and Arg, part of an -R(X3)L/V(X3)E(X3)L- motif, are conserved at these loci throughout this G protein-coupled receptor subfamily and provides molecular insight into key differences between agonist and antagonist binding requirements.
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AVP binding involves interactions with amino acid residues in all three extracellular loop elements and residues in the N-terminus proximal to the first TM segment.
“AVP binding, in contrast, involves interactions with amino acid residues in all three extracellular loop elements and residues in the N-terminus proximal to the first TM segment. (Hawtin et al., 2005; Thibonnier et al., 2000).”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Hawtin et al., 2005
The substitution in marmosets at V45E is adjacent to the highly conserved Arg46.
“Additionally, the substitution in marmosets at V45E is adjacent to the highly conserved Arg46 (Hawtin et al., 2005).”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Hawtin et al., 2005
Many changes in the V1aR N-terminus are not in the AVP binding region, which is proximal to the first TM segment.
“It is worth noting that many of the changes in the V1aR N-terminus are not in the AVP binding region, which is proximal to the first TM segment (Hawtin et al., 2005; Thibonnier et al., 2000).”
From Neuropeptide diversity and the regulation of social behavior in New World primates (2016) · cites this paper as Hawtin et al., 2005
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- Authors
- Hawtin, S. R., Wesley, V. J., Simms, J., Argent, C. C. H., Latif, K., & Wheatley, M.
- Affiliations
- University of Birmingham (GB)
- Corresponding author
- Wheatley, Mark
- Venue
- Molecular Endocrinology
- Publisher
- The Endocrine Society
- Published
- 2005-11-01
- Volume
- 19
- Issue
- 11
- Pages
- 2871-2881
- ISSN
- 0888-8809 (ISSN-L)
- 1944-9917
- OpenAlex
- W2097353938
- Cited by
- 24
- References
- 38
- Citations by year
- 2021: 2 · 2016: 1 · 2014: 1 · 2013: 2 · 2012: 1
- FWCI
- 1.51
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Molecular Endocrinology (publishedVersion)
- PubMed (publishedVersion)
- University of Birmingham Research Portal (University of Birmingham) (submittedVersion)
- Pure (Coventry University) (submittedVersion)
- Funders
- Sanofi
- Wellcome Trust
- Stockholms Universitet
- Directorate for Biological Sciences
- Biotechnology and Biological Sciences Research Council
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Keywords
- vasopressin V1a receptor, receptor signaling, G protein-coupled receptors, alanine scanning, charged residues
- MeSH terms
- Amino Acid Sequence, Animals, Arginine, Arginine Vasopressin, Cells, Cultured, Humans, Models, Molecular, Molecular Sequence Data, Signal Transduction, Mutagenesis, Protein Structure, Secondary, Receptors, Vasopressin, Glutamic Acid, Amino Acid Substitution, Amino Acid Motifs
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