Molecular Basis for Selectivity of High Affinity Peptide Antagonists for the Gastrin-releasing Peptide Receptor

Tokita, Kenji; Katsuno, Tatsuro; Hocart, Simon J.; Coy, David H.; Llinares, Muriel; Martinez, Jean; Jensen, Robert T.

doi:10.1074/jbc.m104566200

Cited by 45 publications

(58 citation statements)

References 70 publications

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“…Studies were performed in transiently transfected Balb-3T3 cells as described in Table 1 binding site conformation. Our results showing a synergetic effect on agonist affinity of multiple substitutions are similar to findings reported for the selectivity of peptide histidine isoleucinamide for the human vasoactive intestinal peptide receptor 1 (Couvineau et al, 1996), GRP for GRPR over human BRS-3 (Nakagawa et al, 2005), the BRS-3-selective agonist Ac-Phe-Trp-Ala-His(tBzl)-Nip-Gly-Arg-NH 2 for BRS-3 over GRPR (Gonzalez et al, 2008), and the peptide antagonists JMV594/JMV641 for GRPR over NMBR (Tokita et al, 2001b) and the nonpeptide antagonist CP96345 for human over the rat neurokinin-1 receptor (Fong et al, 1992). Our finding of the importance of an isoleucine in NMBR instead of serine in GRPR in TM5 (Ile 216 NMBR) or Ala in EC3 (Ile 199 ) has both similarities and differences from findings with a number of other G-protein-coupled receptors (Greenfeder et al, 1999).…”

Section: Tablesupporting

confidence: 70%

“…In contrast, differences in EC2 are primarily important for the high affinity of some BRS-3-selective peptide agonists (Gonzalez et al, 2008), and differences in EC4 are important for GRPR selectivity of the highly selective GRPR peptide antagonists, JMV641 and JMV594 (Tokita et al, 2001b). Likewise, the high affinity of GRP or Bn for GRPR is primarily determined by differences from other Bn receptors in the EC3 region Tokita et al, 2002).…”

Section: Discussionmentioning

confidence: 91%

“…Despite the importance of NMBR in mediating many physiological/pathological processes (Sun et al, 2000;Jensen et al, 2008), there is little known about the molecular basis for its selectivity. This is in marked contrast to the interaction of GRP with GRPR, which has been extensively studied Tokita et al, 2001b;Nakagawa et al, 2005;Jensen et al, 2008). One study ) that used comparative amino acid analysis of the structures of various Bn receptors identified four amino acids (Arg 127 , Ser 205 , His 294 , and Ser 315 ) in BRS-3, which determined its low NMB affinity.…”

Section: Discussionmentioning

confidence: 92%

“…Compared with other G-protein-coupled receptors, our results are similar to the interaction of the peptide agonists, substance P with NK-1 receptors (Li et al, 1996) and CCK-8 with CCK-B receptors (Silvente-Poirot et al, 1998), where both the extracellular and TM domains play a marked role in selectivity. However, the affinity of CCK-8 for the CCK-A receptor (Silvente-Poirot et al, 1998), secretin for secretin receptors (Holtmann et al, 1995), or the GRPR peptide antagonist JMV641 (Tokita et al, 2001b) for GRPR is determined by extracellular domains, whereas differences in only TM regions are crucial for affinity of nociceptin for orphanin-FQ receptors (Meng et al, 1996) or the peptoid antagonist PD168368 for NMBR (Tokita et al, 2001a).…”

Section: Discussionmentioning

confidence: 99%

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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: 70%

Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

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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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“…T3.33 [118] and Y5. 61[268] are located near the polyene tail, but the major charged group associated with the retinal is E3.28 [113]. This residue is located near the Schiff base linkage of the chromophore with K7.…”

Section: Chromophore Conformation and Binding Sitementioning

confidence: 99%

The Crystallographic Model of Rhodopsin and Its Use in Studies of Other G Protein–Coupled Receptors

Filipek

Teller

Palczewski

et al. 2003

Annu. Rev. Biophys. Biomol. Struct.

122

101

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G protein-coupled receptors (GPCRs) are integral membrane proteins that respond to environmental signals and initiate signal transduction pathways activating cellular processes. Rhodopsin is a GPCR found in rod cells in retina where it functions as a photopigment. Its molecular structure is known from cryo-electron microscopic and X-ray crystallographic studies, and this has reshaped many structure/function questions important in vision science. In addition, this first GPCR structure has provided a structural template for studies of other GPCRs, including many known drug targets. After presenting an overview of the major structural elements of rhodopsin, recent literature covering the use of the rhodopsin structure in analyzing other GPCRs will be summarized. Use of the rhodopsin structural model to understand the structure and function of other GPCRs provides strong evidence validating the structural model.

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Analysis of the gastrin‐releasing peptide receptor gene in Italian patients with autism spectrum disorders

Seidita

Mirisola

D'Anna

et al. 2008

American J of Med Genetics Pt B

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The gastrin-releasing peptide receptor (GRPR) was implicated for the first time in the pathogenesis of Autism spectrum disorders (ASD) by Ishikawa-Brush et al. [Ishikawa-Brush et al. (1997): Hum Mol Genet 6: 1241-1250]. Since this original observation, only one association study [Marui et al. (2004): Brain Dev 26: 5-7] has further investigated, though unsuccessfully, the involvement of the GRPR gene in ASD. With the aim of contributing further information to this topic we have sequenced the entire coding region and the intron/exon junctions of the GRPR gene in 149 Italian autistic patients. The results of this study led to the identification of four novel point mutations, two of which, that is, C6S and L181F, involve amino acid changes identified in two patients with ASD and Rett syndrome, respectively. Both the leucine at position 181 and the cysteine at position 6 are strongly conserved in vertebrates. C6S and L181F mutant proteins were expressed in COS-7 and BALB/3T3 cells, but they did not affect either GRP's binding affinity or its potency for stimulating phospholipase C-mediated production of inositol 1,4,5-trisphosphate. In summary, our results do not provide support for a major role of the GRPR gene in ASD in the population of patients we have studied. However, there is a potential role of C6S and L181F mutations on GRPR function, and possibly in the pathogenesis of the autistic disorders in the two patients.

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Molecular Basis for Selectivity of High Affinity Peptide Antagonists for the Gastrin-releasing Peptide Receptor

Cited by 45 publications

References 70 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

The Crystallographic Model of Rhodopsin and Its Use in Studies of Other G Protein–Coupled Receptors

Analysis of the gastrin‐releasing peptide receptor gene in Italian patients with autism spectrum disorders

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