Cloning and Functional Characterization of a Complementary DNA Encoding the Murine Fibroblast Bombesin/Gastrin-Releasing Peptide Receptor

Spindel, Eliot R.; Giladi, Eliezer; Brehm, Paul; Goodman, Richard H.; Segerson, Thomas P.

doi:10.1210/mend-4-12-1956

Cited by 242 publications

(125 citation statements)

References 52 publications

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“…Key Words: bombesin/GRP, MAP kinase, Swiss 3T3, nerve growth factor, epidermal growth factor Bombesin/gastrin-releasing peptide (GRP) and epidermal growth factor (EGF) receptors exert their mitogenic effects in Swiss 3T3 cells by distinct pathways [1]. A cDNA for the bombesin/GRP receptor has been cloned [2,3], and the protein has been shown to have a transmembrane topology like that of other G protein-coupled receptors. Although tyrosine kinase activity has been associated with the receptor function as reported by Isacke et al .…”

Section: Discussionmentioning

confidence: 99%

Bombesin/GRP Stimulates a Protein Kinase in Swiss 3T3 Cells That Phosphorylates Microtubule-Associated Protein Kinase.

Miyasaka

Hayashi

1992

J. Clin. Biochem. Nutr.

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SummaryNerve growth factor (NGF)-and epidermal growth factor (EGF)-stimulated MAP kinase activities from PC-12 cells were recently found to be resolvable into two peaks (microtubule-associated protein[MAP] kinases I and II) by ion-exchange or hydrophobic-interaction HPLC, and both kinases I and II were activated by various growth factors, by okadaic acid, and by phorbol esters. In the present study both MAP kinases I and II were also activated by bombesin/gastrin-releasing peptide (GRP) in Swiss 3T3 cells, and the peak of activation in response to GRP occurred earlier than that observed with EGF. In this work, we confirmed the results reported by Bierman et al. (J. Cell. Phys., 142,[441][442][443][444][445][446][447][448]. Since the cDNA for the bombesin/GRP receptor indicates that this receptor has a transmembrane topology like that of other G-proteincoupled receptors, an alternative signal transduction pathway may mediate the activation of MAP kinase by bombesin/GRP. Key Words: bombesin/GRP, MAP kinase, Swiss 3T3, nerve growth factor, epidermal growth factor Bombesin/gastrin-releasing peptide (GRP) and epidermal growth factor (EGF) receptors exert their mitogenic effects in Swiss 3T3 cells by distinct pathways [1]. A cDNA for the bombesin/GRP receptor has been cloned [2,3], and the protein has been shown to have a transmembrane topology like that of other G protein-coupled receptors. Although tyrosine kinase activity has been associated with the receptor function as reported by Isacke et al . [4], there is no tyrosine kinase homology in the receptor sequence. To evaluate the molecular

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Section: Discussionmentioning

confidence: 99%

Bombesin/GRP Stimulates a Protein Kinase in Swiss 3T3 Cells That Phosphorylates Microtubule-Associated Protein Kinase.

Miyasaka

Hayashi

1992

J. Clin. Biochem. Nutr.

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“…Both GRP and NMB regulate feeding, because treatment with these peptides, as well as BN, decreased the amount of metabolite consumed (4 -6). Meanwhile, five types of BN receptors have been cloned, namely the GRP receptor (GRP-R) (7,8), NMB receptor (NMB-R) (9), BN receptor subtype-3 (BRS-3) (10), BB4 (11), and BRS-3.5 (12). Endogenous ligands for BRS-3 and BRS-3.…”

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confidence: 99%

Leptin Resistance and Enhancement of Feeding Facilitation by Melanin-Concentrating Hormone in Mice Lacking Bombesin Receptor Subtype-3

2004

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Mice lacking either bombesin receptor subtype (BRS)-3 or gastrin-releasing peptide receptor (GRP-R) exhibit feeding abnormalities. However, it is unclear how these receptors are associated with feeding regulation. In BRS-3-deficient mice, we found hyperphagia, subsequent hyperleptinemia, and brain leptin resistance that occurred after the onset of obesity. To explore the cause of this phenomenon, we examined changes in feeding responses to appetite-related neuropeptides in BRS-3-deficient, GRP-R-deficient, and wild-type littermate mice. Among orexigenic neuropeptides, the hyperphagic response to melanin-concentrating hormone (MCH) was significantly enhanced in BRS-3-deficient mice but not in GRP-R-deficient mice. In addition, the levels of MCH-R and prepro-MCH mRNAs in the hypothalamus of BRS-3-deficient mice were significantly more elevated than those of wild-type littermates. There was no significant difference in feeding between BRS-3-deficient and wild-type littermate mice after treatment with bombesin (BN), although the hypophagic response to low-dose BN was significantly suppressed in the GRP-R-deficient mice. These results suggest that upregulation of MCH-R and MCH triggers hyperphagia in BRS-3-deficient mice. From these results, we assume that the BRS-3 gene deletion upsets the mechanism by which leptin decreases the expression of MCH-R and that this effect may be mediated through neural networks independent of BN-related peptides such as GRP-R. Diabetes 53: 570 -576, 2004

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“…To date, two bombesin-like peptides have been isolated from mammalian tissue: NMB 1 in the ranatensin subfamily (3) and GRP in the bombesin subfamily (4). Bombesin, NMB, and GRP bind to a family of G-proteincoupled receptors known to include at least four receptor subtypes: the GRP-preferring receptor (GRP-R (or bb2)) (5,6), the NMB-preferring receptor (NMB-R (or bb1)) (5, 7), bombesin receptor subtype 3 (BRS-3, (or bb3)) (8,9), and bombesin receptor subtype 4 (bb4) (10). Although the four subtypes share about 50% identity when their primary amino acid sequences are aligned, the pharmacology of BRS-3 is significantly different from the pharmacology of the others.…”

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confidence: 99%

Identification of Four Amino Acids in the Gastrin-releasing Peptide Receptor That Are Required for High Affinity Agonist Binding

Akeson¹,

Sainz²,

Mantey³

et al. 1997

Journal of Biological Chemistry

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The bombesin family of G-protein-coupled receptors includes the gastrin-releasing peptide receptor (GRP-R), the neuromedin B receptor (NMB-R), bombesin receptor subtype 3 (BRS-3), and bombesin receptor subtype 4 (bb4). All species homologues of GRP-R, NMB-R, and bb4 bind bombesin with dissociation constants in the nanomolar range; by comparison, human BRS-3 binds bombesin at much lower affinity (K d > 1 M). We used this difference to help identify candidate residues that were potentially critical for forming the bombesin binding pocket. We reasoned that amino acids essential for bombesin binding would be conserved among all homologues of bb4, NMB-R, and GRP-R; conversely, at least one of these amino acids would not be conserved among homologues of BRS-3. Amino acid sequence alignment revealed nine residues that fit this model. We replaced each of these amino acids in mouse GRP-R with the homologous amino acid in human BRS-3. Four substitutions resulted in a significant decrease in bombesin affinity (R288H, Q121R, P199S, and A308S). The analogous mutations in BRS-3 (R127Q, H294R, S205P, and S315A) together resulted in a receptor with a 100-fold increase in bombesin and GRP affinities relative to wildtype BRS-3. From this, we propose a preliminary map of some of the amino acids comprising the agonist binding pocket.Bombesin is a tetradecapeptide that was originally isolated from frog skin (1). Numerous polypeptides with structurally related carboxyl termini were subsequently isolated from amphibians and classified into three subfamilies (bombesin, ranatensin, and phyllolitorin) based upon the sequence of the last three residues in their amidated carboxy-terminal domains (2). To date, two bombesin-like peptides have been isolated from mammalian tissue: NMB 1 in the ranatensin subfamily (3) and GRP in the bombesin subfamily (4). Bombesin, NMB, and GRP bind to a family of G-proteincoupled receptors known to include at least four receptor subtypes: the GRP-preferring receptor (GRP-R (or bb2)) (5, 6), the NMB-preferring receptor (NMB-R (or bb1)) (5, 7), bombesin receptor subtype 3 (BRS-3, (or bb3)) (8, 9), and bombesin receptor subtype 4 (bb4) (10). Although the four subtypes share about 50% identity when their primary amino acid sequences are aligned, the pharmacology of BRS-3 is significantly different from the pharmacology of the others.2 Notably, GRP-R, NMB-R, and bb4 all bind bombesin with affinities in the nanomolar range, whereas BRS-3 binds bombesin with much lower affinity (Ͼ10 M).We hypothesized that amino acids essential for high affinity bombesin binding would be conserved among all known GRP-R, NMB-R, and bb4 sequences and would diverge in the BRS-3 sequences. Nine amino acids in extracellular loops or in the outer half of transmembrane helices fulfilled these conditions. The importance of each of these residues for agonist binding to GRP-R was determined by using site-directed mutagenesis to exchange the residue found in GRP-R with its divergent counterpart in BRS-3. The affinities of BRS-3 or GRP-R mutan...

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Cloning and Functional Characterization of a Complementary DNA Encoding the Murine Fibroblast Bombesin/Gastrin-Releasing Peptide Receptor

Cited by 242 publications

References 52 publications

Bombesin/GRP Stimulates a Protein Kinase in Swiss 3T3 Cells That Phosphorylates Microtubule-Associated Protein Kinase.

Bombesin/GRP Stimulates a Protein Kinase in Swiss 3T3 Cells That Phosphorylates Microtubule-Associated Protein Kinase.

Leptin Resistance and Enhancement of Feeding Facilitation by Melanin-Concentrating Hormone in Mice Lacking Bombesin Receptor Subtype-3

Identification of Four Amino Acids in the Gastrin-releasing Peptide Receptor That Are Required for High Affinity Agonist Binding

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