Design, Synthesis and Some uses of Receptor-Specific Agonists and Antagonists of Vasopressin and Oxytocin

Manning, Maurice; Sawyer, Wilbur H.

doi:10.3109/10799899309073655

Cited by 91 publications

(81 citation statements)

References 36 publications

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“…Since a vast number of peptidic analogues of oxytocin have been developed over the last 40 years (38,39), it would be very interesting to evaluate the structural-functional relationships leading to biased agonist properties. The OTR has been subjected to molecular modeling by several groups (40), and recently, the atosiban-OTR interactions leading to putative high affinity binding have been investigated (41).…”

Section: Discussionmentioning

confidence: 99%

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

Reversi

Rimoldi

Marrocco

et al. 2005

Journal of Biological Chemistry

114

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In human myometrial cells, the promiscuous coupling of the oxytocin receptors (OTRs) to G q and G i leads to contraction. However, the activation of OTRs coupled to different G protein pathways can also trigger opposite cellular responses, e.g. OTR coupling to G i inhibits, whereas its coupling to G q stimulates, cell proliferation. WAF1/CIP1 induction, the same signaling events leading to oxytocin-mediated cell growth inhibition via a G i pathway. Finally, atosiban exposure did not cause OTR internalization and led to only a modest decrease (20%) in the number of high affinity cell membrane OTRs, two observations consistent with the finding that atosiban did not lead to any desensitization of the oxytocin-induced activation of the G q -phospholipase C pathway. Taken together, these observations indicate that atosiban acts as a "biased agonist" of the human OTRs and thus belongs to the class of compounds capable of selectively discriminating only one among the multiple possible active conformations of a single G protein-coupled receptor, thereby leading to the selective activation of a unique intracellular signal cascade.

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Section: Discussionmentioning

confidence: 99%

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

Reversi

Rimoldi

Marrocco

et al. 2005

Journal of Biological Chemistry

114

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“…These peptide receptors share a high primary sequence homology, but display a great diversity in their functional properties. Moreover, these receptors interact with many potent selective agonists and several classes of antagonist ligands, such as cyclic and linear peptides (2) as well as nonpeptides (3,4). By a combination of receptor three-dimensional molecular modeling and site-directed mutagenesis approaches, our preceding study has led to the mapping of the AVP agonist-binding domain in the V1a receptor subtype (5).…”

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confidence: 99%

Identification of Residues Responsible for the Selective Binding of Peptide Antagonists and Agonists in the V2 Vasopressin Receptor

Cotte¹,

Balestre²,

Phalipou³

et al. 1998

Journal of Biological Chemistry

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To improve our understanding of the functional architecture of G protein-coupled receptors, we have taken advantage of differences among mammalian species in ligand binding to search for the rat versus human selectivity determinants of the V2 vasopressin receptor and of its peptide ligands. Our data indicate that residue 2 of species-selective peptide antagonists such as d(CH 2 ) 5 -[DIle 2 ,Ile 4 ,Tyr-NH 2 9 ]arginine vasopressin controls their rat versus human selectivity. For species-selective agonists such as desmopressin, residues 1 and 8 modulate the binding selectivity. Among residues different between rat and human V2 receptors, those localized in the upper part of the human V2 receptor have been substituted with their rat V2 homologs. Pharmacological analysis of mutant receptors revealed that residues 202 and 304 fully control the species selectivity of the discriminating antagonists in an independent and additive manner. A third residue (position 100) is necessary to observe an equivalent phenomenon for the discriminating agonists. The substitution of these three residues does not modify the affinity of the nonselective agonists and antagonists. In conclusion, extracellular loops and the top of the transmembrane domains of V2 vasopressin receptors may provide the molecular basis for peptide ligand-binding species selectivity. Very few residues in these regions may control the binding mode of both agonists and antagonists. The arginine vasopressin (AVP)1 /oxytocin receptor family represents a suitable system to investigate structure-function relationships among G protein-coupled receptors. Indeed, all four receptor subtypes for these neurohypophyseal hormones, namely the V1a, V1b, and V2 AVP receptors and the unique oxytocin receptor, have been well characterized by pharmacological and molecular cloning studies (see Ref. 1 for review). These peptide receptors share a high primary sequence homology, but display a great diversity in their functional properties. Moreover, these receptors interact with many potent selective agonists and several classes of antagonist ligands, such as cyclic and linear peptides (2) as well as nonpeptides (3,4). By a combination of receptor three-dimensional molecular modeling and site-directed mutagenesis approaches, our preceding study has led to the mapping of the AVP agonist-binding domain in the V1a receptor subtype (5). The hormone-binding site is localized in a narrow cleft delimited by most of the transmembrane regions ϳ15 Å away from the extracellular surface. Because of the high conservation of residues involved in the interaction with the hormone, this binding site has been proposed to be common to the V1b, V2, and oxytocin receptors. An extracellular residue responsible for receptor subtype agonist selectivity has also been identified (6 -8).These first analyses did not provide much information on the definition of AVP/oxytocin receptor antagonist-binding domains. To date, only the upper part of transmembrane region VII and a hydrophobic cluster of aromatic residue...

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“…These receptors are able to specifically bind the natural hormones or synthetic peptide analogues by distinguishing subtle structural differences in their sequence (Akhundova et al, 1996). In addition, numerous agonist and antagonist ligands have been described over 40 years for this receptor family (Manning and Sawyer, 1993). Thus, these membrane proteins represent a good model for the study of the molecular mechanisms involved in hormone-receptor interactions.…”

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confidence: 99%

“…F-34094 Montpellier Cedex 5, France (Manning and Sawyer, 1993). Thus, these membrane proteins represent a good model for the study of the molecular mechanisms involved in hormone-receptor interactions.…”

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confidence: 99%

Efficient Photoaffinity Labeling of the Rat V_1a, Vasopressin Receptor using a Linear Azidopeptidic Antagonist

Carnazzi

Aumelas

Phalipou

et al. 1997

European Journal of Biochemistry

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We have synthesized and fully characterized by fast-atom-bombardment-mass, NMR and ultraviolet spectroscopies the vasopressin antagonist 3-azidophenylpropionyl-~-Tyr(Me)-Phe-Gln-Asn-Arg-Pro-ArgTyr(31)-NH2. Easily radioiodinatable just before use, it has a high affinity for the natural rat liver V,, receptor [dissociation constant (Kd) = 54 2 20 pM; Carnazzi, E., Aumelas, A., Barberis, C., Guillon, G. & Seyer, R. (1994) J. Med. Chem. 37, 1841-18491 and for both the rat vasopressin V,, receptor expressed in Sixx.iopteru frugiperdu 9 cells (Sf9 cells, Kd = 688 i 35 pM) and in COS-7 cells (Kd = 320 2 20 pM). This probe labels specifically the V,, receptors in an ultraviolet-dependent manner, and binds covalently to about 12% of the receptors with high stability over several days, even in dissociation or solubilization conditions. SDS/PAGE studies and autoradiographic analyses of the photolabeled receptors reveal a single band (49.5 kDa) and two bands (63 kDa and 93.6 kDa) for receptor-probe associations obtained in Sf9 and COS-7 cells respectively. These molecular masses are consistent with non-glycosylated and highly glycosylated forms of the receptor, according to each expression system. In rat liver membranes, we have identified apparent molecular masses of about 32, 45 and more than 67 kDa. We finally demonstrated a proteolysis of the receptor that appeared to be Znz+ and leupeptin sensitive. The high potency of this ligand is promising for the monitoring of the purification of the V,, receptor and for mapping its antagonist-binding site.Keywords: photoaffinity ; vasopressin; antagonist; receptor; proteolysis.Vasopressin and oxytocin are two neurohypophyseal hormones that exert a wide range of physiological effects upon binding to different guanine-nucleotide-binding-regulatory(G)-protein-coupled receptor subtypes (Jard, 1983). More than a dozen of these oxytocin and vasopressin receptor subtypes have been cloned in several species, among which the vasopressin V,, subtype cloned in the rat (Morel et al., 1992;Innamorati et al., 1995), the sheep (Hutchins et al., 1995) and the human (Thibonnier et al., 1994). One of the most recently cloned members of this family is the mesotocin receptor, described by Akhundova et al. (1996). These receptors are able to specifically bind the natural hormones or synthetic peptide analogues by distinguishing subtle structural differences in their sequence (Akhundova et al., 1996). In addition, numerous agonist and antagonist ligands have been described over 40 years for this receptor family (Manning and Sawyer, 1993). Thus, these membrane proteins represent a good model for the study of the molecular mechanisms involved in hormone-receptor interactions.The analysis of the hydrophobicity profile of these receptor sequences confirms that they belong to the seven-transmembrane-domain receptor family. The comparison of the amino acid composition of all the known members of this family has revealed the presence of conserved residues and led to some structure/activity presumptions. A...

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Design, Synthesis and Some uses of Receptor-Specific Agonists and Antagonists of Vasopressin and Oxytocin

Cited by 91 publications

References 36 publications

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

Identification of Residues Responsible for the Selective Binding of Peptide Antagonists and Agonists in the V2 Vasopressin Receptor

Efficient Photoaffinity Labeling of the Rat V_1a, Vasopressin Receptor using a Linear Azidopeptidic Antagonist

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Design, Synthesis and Some uses of Receptor-Specific Agonists and Antagonists of Vasopressin and Oxytocin

Cited by 91 publications

References 36 publications

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

The Oxytocin Receptor Antagonist Atosiban Inhibits Cell Growth via a “Biased Agonist” Mechanism

Identification of Residues Responsible for the Selective Binding of Peptide Antagonists and Agonists in the V2 Vasopressin Receptor

Efficient Photoaffinity Labeling of the Rat V1a, Vasopressin Receptor using a Linear Azidopeptidic Antagonist

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Efficient Photoaffinity Labeling of the Rat V_1a, Vasopressin Receptor using a Linear Azidopeptidic Antagonist