Critical Role of a Subdomain of the N-Terminus of the V 1 a Vasopressin Receptor for Binding Agonists but Not Antagonists; Functional Rescue by the Oxytocin Receptor N-Terminus
Abstract
A fundamental issue in molecular pharmacology is to define how agonist:receptor interaction differs from that of antagonist:receptor. The V(1a) receptor (V(1a)R) is a member of a family of related G-protein-coupled receptors that are activated by the neurohypophysial peptide hormone arginine-vasopressin (AVP). Here we define a short subdomain of the N-terminus of the V(1a)R from Glu(37) to Asn(47) that is an absolute requirement for binding AVP and other agonists. In marked contrast to the situation for agonists, deleting this segment has little or no effect on the binding of either peptide or non-peptide antagonists. In addition, we established that this subdomain was crucial for receptor activation and second messenger generation. The oxytocin receptor (OTR) also binds AVP with high affinity but exhibits a different pharmacological profile to the V(1a)R. Substitution of the N-terminus of the V(1a)R with the corresponding sequence from the OTR generated a chimeric receptor (OTR(N)-V(1a)R). The presence of the OTR N-terminus recovered high affinity agonist binding such that the OTR(N)-V(1a)R possessed almost wild-type V(1a)R pharmacology and signaling. Consequently, a domain within the N-terminus is required for agonist binding but it does not provide the molecular discriminator for subtype-selective agonist recognition. Cotransfection and peptide mimetic studies demonstrated that this N-terminal subdomain had to be contiguous with the receptor polypeptide to be functional. This study establishes that a segment of the V(1a)R N-terminus has a pivotal role in the mechanism of agonist binding and provides molecular insight into key differences between the interaction of agonists and antagonists with a peptide receptor family.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
AVP has similar affinity for OTRs, V1aRs, and V1bRs, whereas OT may have a higher affinity for OTRs than for V1aRs or V1bRs.
“AVP has a similar affinity for OTRs, V1aRs, and V1bRs while OT may have a higher affinity for OTRs than V1aRs or V1bRs [28; 29; 30; 31; 32; 33].”
From Cross-talk among oxytocin and arginine-vasopressin receptors: Relevance for basic and clinical studies of the brain and periphery (2018) · cites this paper as [29]
There can be differences of around 10-100 fold in the selectivity of OTRs and AVP receptors for OT and AVP.
“Nevertheless, it appears that there can be differences of around 10–100 fold in the selectivity of OTRs and AVP receptors for OT and AVP.”
From Cross-talk among oxytocin and arginine-vasopressin receptors: Relevance for basic and clinical studies of the brain and periphery (2018) · cites this paper as [29]
Cites
Hulme, E. C., Birdsall, N. J. M., & Buckley, N. J. (1990) · The Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev.pa.30.040190.003221 · Cited by 1,344 · see citing works
STRUCTURE AND FUNCTION OF G PROTEIN-COUPLED RECEPTORS
Strader, C. D., Fong, T. M., Tota, M. R., Underwood, D. J., & Dixon, R. A. F. (1994) · Annual Review of Biochemistry · doi:10.1146/annurev.bi.63.070194.000533 · Cited by 1,080 · see citing works
The probable arrangement of the helices in G protein‐coupled receptors.
Baldwin, J. M. (1993) · The EMBO Journal · doi:10.1002/j.1460-2075.1993.tb05814.x · Cited by 968 · see citing works
Kruszynski, M., Lammek, B., Manning, M., Seto, J., Haldar, J., & Sawyer, W. H. (1980) · Journal of Medicinal Chemistry · doi:10.1021/jm00178a003 · Cited by 450 · see citing works
Michell, R. H., Kirk, C. J., & BILLAH, M. M. (1979) · Biochemical Society Transactions · doi:10.1042/bst0070861 · Cited by 440 · see citing works
Gal, C. S., Wagnon, J., Garcia, C., Lacour, C., Guiraudou, P., Christophe, B., Villanova, G., Nisato, D., Maffrand, J., & Fur, G. L. (1993) · Journal of Clinical Investigation · doi:10.1172/jci116554 · Cited by 272 · see citing works
Scheven, B. A. A., Visser, J. W. M., & Nijweide, P. J. (1986) · Nature · doi:10.1038/321079a0 · Cited by 267 · see citing works
The Binding Site of Neuropeptide Vasopressin V1a Receptor
Mouillac, B., Chini, B., Balestre, M.-N., Elands, J., Trumpp-Kallmeyer, S., Hoflack, J., Hibert, M., Jard, S., & Barberis, C. (1995) · Journal of Biological Chemistry · doi:10.1074/jbc.270.43.25771 · Cited by 262 · see citing works
Tyr115 is the key residue for determining agonist selectivity in the V1a vasopressin receptor
Chini, B., Mouillac, B., Ala, Y., Balestre, M. N., Trumpp‐Kallmeyer, S., Hoflack, J., Elands, J., Hibert, M., Manning, M., & Jard, S. (1995) · The EMBO Journal · doi:10.1002/j.1460-2075.1995.tb07211.x · Cited by 179 · see citing works
Postina, R., Kojro, E., & Fahrenholz, F. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.49.31593 · Cited by 134 · see citing works
The condensing vacuole of exocrine cells is more acidic than the mature secretory vesicle
Orci, L., Ravazzola, M., & Anderson, R. G. W. (1987) · Nature · doi:10.1038/326077a0 · Cited by 128 · see citing works
Intracellular Third Loop Domain of Angiotensin II Type-2 Receptor
Hayashida, W., Horiuchi, M., & Dzau, V. J. (1996) · Journal of Biological Chemistry · doi:10.1074/jbc.271.36.21985 · Cited by 117 · see citing works
Howl, J., Ismail, T., Strain, A. J., Kirk, C. J., Anderson, D., & Wheatley, M. (1991) · Biochemical Journal · doi:10.1042/bj2760189 · Cited by 67 · see citing works
An extracellular residue determines the agonist specificity of V2 vasopressin receptors
Ufer, E., Postina, R., Gorbulev, V., & Fahrenholz, F. (1995) · FEBS Letters · doi:10.1016/0014-5793(95)00150-8 · Cited by 65 · see citing works
Deletions of the N‐terminal regions of the human melanocortin receptors
Schiöth, H. B., Petersson, S., Muceniece, R., Szardenings, M., & Wikberg, J. E. S. (1997) · FEBS Letters · doi:10.1016/s0014-5793(97)00593-0 · Cited by 53 · see citing works
Structure and expression of an ovine endometrial oxytocin receptor cDNA
Riley, P. R., Flint, A. P. F., Abayasekara, D. R. E., & Stewart, H. J. (1995) · Journal of Molecular Endocrinology · doi:10.1677/jme.0.0150195 · Cited by 37 · see citing works
Huguenin, R. L. (1964) · Helvetica Chimica Acta · doi:10.1002/hlca.19640470733 · Cited by 37 · see citing works
Howl, J., & Wheatley, M. (1996) · Biochemical Journal · doi:10.1042/bj3170577 · Cited by 34 · see citing works
Identification of a domain of ETA receptor required for ligand binding
Adachi, M., Yang, Y.-Y., Trzeciak, A., Furuichi, Y., & Miyamoto, C. (1992) · FEBS Letters · doi:10.1016/0014-5793(92)81393-z · Cited by 34 · see citing works
Chimeric strategies for the rational design of bioactive analogs of small peptide hormones
Howl, J., Langel, Ü., Hawtin, S. R., Valkna, A., YARWOOD, N. J., Saar, K., & Wheatley, M. (1997) · The FASEB Journal · doi:10.1096/fasebj.11.7.9212082 · Cited by 32 · see citing works
Howl, J., Wang, X. F., Kirk, C. J., & Wheatley, M. (1993) · European Journal of Biochemistry · doi:10.1111/j.1432-1033.1993.tb17811.x · Cited by 30 · see citing works
Discovery and development of non-peptide antagonists of peptide hormone receptors
Pettibone, D. J., & Freidinger, R. M. (1997) · Biochemical Society Transactions · doi:10.1042/bst0251051 · Cited by 23 · see citing works
Howl, J., New, D. C., & Wheatley, M. (1992) · Journal of Molecular Endocrinology · doi:10.1677/jme.0.0090123 · Cited by 9 · see citing works
Probing the V1a vasopressin receptor binding site with pyroglutamate-substituted linear antagonists
Howl, J., YARWOOD, N. J., Stock, D., & Wheatley, M. (1996) · Neuropeptides · doi:10.1016/s0143-4179(96)90058-x · Cited by 9 · see citing works
Homology between neurohypophyseal hormone receptors
Wheatley, M., Howl, J., Morel, A., & Davies, A. R. L. (1993) · Biochemical Journal · doi:10.1042/bj2960519 · Cited by 9 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Vasopressin V1a and V1b Receptors: From Molecules to Physiological Systems
Koshimizu, T., Nakamura, K., Egashira, N., Hiroyama, M., Nonoguchi, H., & Tanoue, A. (2012) · Physiological Reviews · doi:10.1152/physrev.00035.2011 · Cited by 413 · see citing works
Song, Z., & Albers, H. E. (2018) · Frontiers in Neuroendocrinology · doi:10.1016/j.yfrne.2017.10.004 · Cited by 244 · see citing works
Neurobiology of social attachments
Coria‐Ávila, G. A., Manzo, J., García, L. I., Carrillo, P., Miquel, M., & Pfaus, J. G. (2014) · Neuroscience & Biobehavioral Reviews · doi:10.1016/j.neubiorev.2014.04.004 · Cited by 106 · see citing works
Systematic Analysis of the Entire Second Extracellular Loop of the V1a Vasopressin Receptor
Conner, M. T., Hawtin, S. R., Simms, J., Wootten, D. L., Lawson, Z., Conner, A. C., Parslow, R. A., & Wheatley, M. (2007) · Journal of Biological Chemistry · doi:10.1074/jbc.m702151200 · Cited by 82 · see citing works
Hawtin, S. R., Tobin, A. B., Patel, S. S., & Wheatley, M. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m106142200 · Cited by 74 · see citing works
Curcumin: a new cell-permeant inhibitor of the inositol 1,4,5-trisphosphate receptor
Dyer, J. L., Khan, S., Bilmen, J. G., Hawtin, S. R., Wheatley, M., Javed, M., & Michelangeli, F. (2002) · Cell Calcium · doi:10.1054/ceca.2001.0259 · Cited by 61 · see citing works
Genome-wide prediction of age at puberty and reproductive longevity in sows
Tart, J. K., Johnson, R. K., Bundy, J. W., Ferdinand, N. N., McKnite, A. M., Wood, J. R., Miller, P. S., Rothschild, M. F., Spangler, M. L., Garrick, D. J., et al. (2013) · Animal Genetics · doi:10.1111/age.12028 · Cited by 52 · see citing works
Hawtin, S. R., Simms, J., Conner, M. T., Lawson, Z., Parslow, R. A., Trim, J. E., Sheppard, A., & Wheatley, M. (2006) · Journal of Biological Chemistry · doi:10.1074/jbc.m607639200 · Cited by 45 · see citing works
A Three‐Site Mechanism for Agonist/Antagonist Selective Binding to Vasopressin Receptors
Saleh, N., Saladino, G., Gervasio, F. L., Haensele, E., Banting, L., Whitley, D. C., Santos, J. S. O., Bureau, R., & Clark, T. (2016) · Angewandte Chemie International Edition · doi:10.1002/anie.201602729 · Cited by 40 · see citing works
Hawtin, S. R., DAVIES, A. R. L., Matthews, G. M., & Wheatley, M. (2001) · Biochemical Journal · doi:10.1042/bj3570073 · Cited by 39 · see citing works
Lindner, D., Walther, C., TENNEMANN, A., & Beck‐Sickinger, A. G. (2008) · Cellular Signalling · doi:10.1016/j.cellsig.2008.09.007 · Cited by 34 · see citing works
Kartz, G. A., Holme, R. L., Nicholson, K., & Sahoo, D. (2014) · Biochemistry · doi:10.1021/bi500706x · Cited by 32 · see citing works
Subtype-selective Noncompetitive or Competitive Inhibition of Human α1-Adrenergic Receptors by ρ-TIA
Chen, Z., Rogge, G. A., Hague, C., Alewood, D., Colless, B., Lewis, R. J., & Minneman, K. P. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m403703200 · Cited by 30 · see citing works
Ren, D., Chin, K. R., & French, J. A. (2014) · PLoS ONE · doi:10.1371/journal.pone.0111638 · Cited by 28 · see citing works
Extramembranous Regions in G Protein-Coupled Receptors: Cinderella in Receptor Biology?
Pal, S., & Chattopadhyay, A. (2019) · The Journal of Membrane Biology · doi:10.1007/s00232-019-00092-3 · Cited by 25 · see citing works
Extracellular loops and ligand binding to a subfamily of Family A G-protein-coupled receptors
Wheatley, M., Simms, J., Hawtin, S. R., Wesley, V. J., Wootten, D. L., Conner, M. T., Lawson, Z., Conner, A. C., Baker, A., Cashmore, Y., et al. (2007) · Biochemical Society Transactions · doi:10.1042/bst0350717 · Cited by 24 · see citing works
Hawtin, S. R., Wesley, V. J., Simms, J., Argent, C. C. H., Latif, K., & Wheatley, M. (2005) · Molecular Endocrinology · doi:10.1210/me.2005-0148 · Cited by 24 · see citing works
Self-citationHawtin, S. R., Wesley, V. J., Parslow, R. A., Simms, J., Miles, A., McEwan, K., & Wheatley, M. (2002) · Molecular Endocrinology · doi:10.1210/mend.16.3.0795 · Cited by 24 · see citing works
Lyu, L. K., Li, J. S., Wang, X. J., Yao, Y. J., Li, J. F., Li, Y., Wen, H. S., & Qi, X. (2021) · Frontiers in Endocrinology · doi:10.3389/fendo.2021.617580 · Cited by 20 · see citing works
Calderón, J. C., Plut, E., Keller, M., Cabrele, C., Reiser, O., Gervasio, F. L., & Clark, T. (2023) · Journal of Chemical Information and Modeling · doi:10.1021/acs.jcim.3c01574 · Cited by 15 · see citing works
Hoffmann, C., Soltysiak, K. A., West, P. L., & Jacobson, K. A. (2004) · Biochemical Pharmacology · doi:10.1016/j.bcp.2004.07.014 · Cited by 13 · see citing works
A Three‐Site Mechanism for Agonist/Antagonist Selective Binding to Vasopressin Receptors
Saleh, N., Saladino, G., Gervasio, F. L., Haensele, E., Banting, L., Whitley, D. C., Santos, J. S. O., Bureau, R., & Clark, T. (2016) · Angewandte Chemie · doi:10.1002/ange.201602729 · Cited by 12 · see citing works
Agonist-specific requirement for a glutamate in transmembrane helix 1 of the oxytocin receptor
Wootten, D. L., Simms, J., Massoura, A. J., Trim, J. E., & Wheatley, M. (2010) · Molecular and Cellular Endocrinology · doi:10.1016/j.mce.2010.11.029 · Cited by 10 · see citing works
Yamasue, H. (2023) · Peptides · doi:10.1016/j.peptides.2023.171133 · Cited by 8 · see citing works
7 Oxytocin Antagonists as Potential Therapeutic Agents for the Treatment of Preterm Labour
Allen, M. J., Livermore, D. G., & Mordaunt, J. E. (2006) · Progress in medicinal chemistry · doi:10.1016/s0079-6468(05)44407-0 · Cited by 8 · see citing works
Hawtin, S. R., Wesley, V. J., Simms, J., Parslow, R. A., Miles, A., McEwan, K., Keen, M., & Wheatley, M. (2003) · European Journal of Biochemistry · doi:10.1046/j.1432-1033.2003.03865.x · Cited by 8 · see citing works
Cho, H.-J., Moon, M. J., Kwon, H. B., Hwang, J., & Seong, J. Y. (2007) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2007.11.010 · Cited by 4 · see citing works
Chapter 13 Expression of human vasopressin and oxytocin receptors in Escherichia coli
Mouillac, B., Sen, T., Durroux, T., Gaibelet, G., & Barberis, C. (2002) · Progress in brain research · doi:10.1016/s0079-6123(02)39015-0 · Cited by 3 · see citing works
Trenhaile, M. D. (2015) · Lincoln (University of Nebraska) · Cited by 1 · see citing works
Appendix: Site‐Directed Mutagenesis and Chimeras
Poyner, D. R., & Wheatley, M. (2010) · doi:10.1002/9780470749210.app1 · Cited by 1 · see citing works
Increasing the Odds of Drug Hit Identification by Screening Against Receptor Homologs?
Ji, Z., Li, Z., Wang, J., Cai, C., Han, L., Zheng, C., & Chen, Y. (2006) · Letters in Drug Design & Discovery · doi:10.2174/157018006776286970
Inada, K. (2026) · Neuroscience Research · doi:10.1016/j.neures.2026.105109
- Authors
- Hawtin, S. R., Wesley, V. J., Parslow, R. A., Patel, S., & Wheatley, M.
- Affiliations
- MSD (UK) Limited (United Kingdom) (GB)
- Corresponding author
- Wheatley, Mark
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2000-10-10
- Volume
- 39
- Issue
- 44
- Pages
- 13524-13533
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2009084148
- Cited by
- 32
- References
- 25
- Citations by year
- 2026: 1 · 2023: 2 · 2021: 1 · 2019: 1 · 2017: 1 · 2016: 2 · 2015: 1 · 2014: 3 · 2013: 1 · 2012: 1
- FWCI
- 0.72
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- 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
- Social Sciences › Psychology › Social Psychology › Neuroendocrine regulation and behavior
- Keywords
- vasopressin V1a receptor, oxytocin receptor, agonist binding, antagonist binding, receptor activation, chimeric receptors, vasopressin, G protein-coupled receptors, peptide hormone receptors, second messenger signaling
- MeSH terms
- Amino Acid Sequence, Animals, Arginine Vasopressin, Cell Line, Humans, Ligands, Molecular Sequence Data, Peptide Fragments, Protein Binding, Recombinant Fusion Proteins, Sheep, Transfection, Sequence Deletion, Protein Structure, Tertiary, Receptors, Vasopressin, Receptors, Oxytocin, Rats, Antidiuretic Hormone Receptor Antagonists
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
