The Biologically Crucial C Terminus of Cholecystokinin and the Non-peptide Agonist SR-146,131 Share a Common Binding Site in the Human CCK1 Receptor

Escrieut, Chantal; Gigoux, Véronique; Archer, Elodie; Verrier, Sophie; Maigret, Bernard; Behrendt, Raymond; Moröder, Luis; Bignon, Eric; Silvente-Poirot, Sandrine; Pradayrol, Lucien; Fourmy, Daniel

doi:10.1074/jbc.m108563200

Cited by 58 publications

(109 citation statements)

References 39 publications

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“…Likewise, isoleucine in TM3 of the melanocortin-4 receptor was important for high affinity for melanocortin-4 receptor ligands (tetrahydroisoquinoline, M10, and adrenocorticotropin hormone) (Nickolls et al, 2003). Our results are similar to findings with both the CCK-1 and NK-2 receptors, where replacement of isoleucine in TM5 or TM7, respectively, with alanine markedly decreased agonist affinity (Greenfeder et al, 1999;Escrieut et al, 2002). There is almost no information on the molecular mechanisms that determine the importance of the presence of an isoleucine in the different Gprotein-coupled receptors for high-affinity ligand interaction.…”

Section: Tablesupporting

confidence: 85%

“…However, when isoleucine 199 was substituted by amino acids with high hydrophobicity (leucine and valine) or with a moderately high hydrophobicity (methionine), either no difference or only a slight decrease in the affinity of NMB for NMBR was observed. These results support the conclusion that the hydrophobicity of the substitution in position 199 of NMBR plays a crucial role for NMBR affinity for NMB and suggest that the substitution of isoleucine with alanine creates a difference in the transfer-free energies that could influence hydrophobicity and the thermodynamics of protein folding (Escrieut et al, 2002).…”

Section: Tablesupporting

confidence: 84%

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Molecular Basis for the Selectivity of the Mammalian Bombesin Peptide, Neuromedin B, for Its Receptor

González

Nakagawa²,

Mantey³

et al. 2009

J Pharmacol Exp Ther

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The mammalian bombesin (Bn) peptides, neuromedin B (NMB) and gastrin-releasing peptide (GRP), have widespread actions in many tissues, and their effects are mediated by two closely related G-protein-coupled receptors, the NMBR and GRPR. Little is known about the structural determinants of NMBR selectivity for NMB, in contrast to GRP selectivity for the GRPR, which has been extensively studied. To provide insight, chimeric NMBR-GRPR loss-of-affinity and gain-of-affinity mutants were made, as well as NH 2 -terminally truncated NMBR and point mutants using site-directed mutagenesis. Receptors were expressed in Balb-3T3-cells or CHOP cells, and affinities were determined. NMB had 115-fold greater affinity for NMBR than GRPR. Receptor-chimeric studies showed that NMBR selectivity for NMB was primarily determined by differences in the third extracellular (EC3) regions of GRPR-NMBR and adjacent upper-transmembrane-5 (TM5) region. In this region, 24 NMB gain-of-affinity GRPR mutants or NMBR loss-of-affinity point/ combination mutants were made. Three gain-of-affinity mutant GRPRs [[A198I] (EC3), [H202Q] (EC3), [S215I] (upper TM5)]had increased NMB affinity (2.4 -21-fold), and these results were confirmed with NMBR loss-of-affinity mutants [I199A, Q203H,I215S-NMBR]. The combination mutant [A198I, S215]GRPR had the greatest effect causing a complete NMB gain-of-affinity. The importance of differences at position 199NMBR or 203NMBR was studied by substituting amino acids with various properties. Our results show that NMBR selectivity for NMB is due to differences in the EC3 of NMBR-GRPR and the adjacent upper-TM5 region. Within these regions, isoleucines in NMBR [position 199 (EC3)] (instead of A198GRPR) and in 215NMBR (TM5) (instead of S214GRPR), as well as Q203NMBR (instead of H202GRPR) are responsible for high NMB-affinity/selectivity of NMBR. The effect at position 199 is primarily due to differences in hydrophobicity of the substitution, whereas steric factors and charge of the substitution at position 203 were important determinants of NMB selectivity.

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Section: Tablesupporting

confidence: 85%

Section: Tablesupporting

confidence: 84%

Molecular Basis for the Selectivity of the Mammalian Bombesin Peptide, Neuromedin B, for Its Receptor

González

Nakagawa²,

Mantey³

et al. 2009

J Pharmacol Exp Ther

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“…The NH 2 -terminal moiety of CCK is tightly linked to extracellular residues of CCK1R, including residues in the second extracellular loop at the top of transmembrane (TM) helix I and within the third extracellular loop (16,237). The COOH-terminal tetrapeptide of CCK appears to be embedded between TM helices III, V, VI, and VII through a network of both ionic and hydrophobic interactions (16,145,168). This binding mode of the COOH terminus of CCK into CCK1R is in agreement with an NMR study of the interactions between CCK and a fragment of CCK1R comprising the top portion of helix VI and the third extracellular loop as well as a fragment including amino acids at the top of transmembrane segment I (173, 373).…”

Section: A Localization Of Ligand Binding Sitesmentioning

confidence: 99%

Cholecystokinin and Gastrin Receptors

Dufresne

Seva²,

Fourmy³

2006

Physiological Reviews

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418

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Cholecystokinin and gastrin receptors (CCK1R and CCK2R) are G protein-coupled receptors that have been the subject of intensive research in the last 10 years with corresponding advances in the understanding of their functioning and physiology. In this review, we first describe general properties of the receptors, such as the different signaling pathways used to exert short- and long-term effects and the structural data that explain their binding properties, activation, and regulation. We then focus on peripheral cholecystokinin receptors by describing their tissue distribution and physiological actions. Finally, pathophysiological peripheral actions of cholecystokinin receptors and their relevance in clinical disorders are reviewed.

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“…Several critical contacts between the CCK1R binding site and CCK were successively discovered: Met195 and Arg197 located in the second extracellular loop, were shown to interact with the sulfated tyrosine (Gigoux et al 1998(Gigoux et al & 1999b; Arg336 and Asn333 at the top of helix VI were demonstrated to pair with the Asp carboxylate and the Cterminal amide of CCK, respectively (Gigoux et al 1999a). More recently, search for residues of the receptor in interaction with Met/Nle and Phe of CCK led to description of a network of hydrophobic residues including Cys94, Met121, Val125, Phe218, Ile329, Phe330, Trp326, Ile352, Leu356 and Tyr360 all involved in the binding site for CCK and in the activation process of the CCK1R (Escrieut et al 2002) (fig. 2).…”

Section: Mapping Of Cck1r Binding Site For Cckmentioning

confidence: 99%

“…Therefore, side-chains of Met121 of CCK1R and Met of CCK are not directly involved in the activation process but the ''hydrophobic weight'' near position 121 is crucial for CCK1R activation. Application of dynamic procedures on the model of the CCK1R.CCK complex yielded the very exciting result that introduction of hydrophobic residues near position 121 determines positioning of the aromatic ring of the Phe of CCK within the binding pocket (Escrieut et al 2002). Obviously, this modelling result needs experimental verification.…”

Section: First Structural Data On the Mechanism Of Activation/ Inactimentioning

confidence: 99%

Structure of Cholecystokinin Receptor Binding Sites and Mechanism of Activation/Inactivation by Agonists/Antagonists

Fourmy

Escrieut

Archer

et al. 2002

Pharmacology & Toxicology

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Delineation of CCK receptor binding sites is a prerequisite for the understanding of the molecular basis for ligand recognition, partial agonism, ligand-induced traffiking of receptor signalling. In the current paper, we illustrate how, in the past 5 years, studies from our laboratory and others have provided new data on the molecular basis of the pharmacology and functioning of CCK1 and CCK2 receptors. Available data on CCK1 and CCK2R binding sites indicate that 1) homologous regions of the two receptors are involved in the binding site of CCK, however, positioning of CCK slightly differs; 2) binding sites of non-peptide agonists/ antagonist are buried in the pocket formed by transmembrane helices and overlap that of CCK and 3) residues of the binding sites as well as of conserved motifs such as E/DRY, NPXXY are crucial for receptor activation.The actions of cholecystokinin (CCK) and gastrin are mediated by specific high affinity membrane receptors on target cells. These receptors have been characterized pharmacologically and biologically and are divided into two subtypes based on their affinities for CCK and gastrin. The CCK1 receptor (previously named CCK-A receptor) has an approximately 500 times higher affinity for CCK than for gastrin and the CCK2 receptor (previously named CCKB/ G receptor) has the same high affinity for both CCK and gastrin. An additional difference between the CCK1 and CCK2 receptors resides in their affinities for sulfated and non-sulfated CCK. Indeed, whereas the CCK1 receptor binds and responds to sulfated CCK with 500 times higher affinity/potency than non-sulfated CCK, the CCK2 receptor weakly discriminates between the two (Silvente-Poirot et al. 1993;Wank 1998).

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The Biologically Crucial C Terminus of Cholecystokinin and the Non-peptide Agonist SR-146,131 Share a Common Binding Site in the Human CCK1 Receptor

Cited by 58 publications

References 39 publications

Molecular Basis for the Selectivity of the Mammalian Bombesin Peptide, Neuromedin B, for Its Receptor

Molecular Basis for the Selectivity of the Mammalian Bombesin Peptide, Neuromedin B, for Its Receptor

Cholecystokinin and Gastrin Receptors

Structure of Cholecystokinin Receptor Binding Sites and Mechanism of Activation/Inactivation by Agonists/Antagonists

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