Discovery of a high affinity radioligand for the human orphan receptor, bombesin receptor subtype 3, which demonstrates that it has a unique pharmacology compared with other mammalian bombesin receptors

Samuel A. Mantey, H. Christian Weber, Eduardo Sainz, Mark Akeson, Richard R. Ryan, Tapas K. Pradhan, Robert Searles, Eliot Spindel, James F. Battey, David H. Coy, Robert T. Jensen

    Research output: Contribution to journalArticle

    137 Citations (Scopus)

    Abstract

    An orphan receptor discovered in 1993 was called bombesin receptor subtype 3 (BRS-3) because of 47-51% amino acid identity with bombesin (Bn) receptors. Its pharmacology is unknown, because no naturally occurring tissues have sufficient receptors to allow studies. We made two cell lines stably expressing the human BRS-3 (hBRS-3). hBRS-3 was overexpressed in the human non-small cell lung cancer cells, NCI-H1299, and the other was made in Balb 3T3 cells, which lack endogenous BRS-3. [D-Phe6,β- Ala11,Phe13,Nle14]Bn-(6-14) (where Nle represents norleucine) was discovered to have high potency for stimulating inositol phosphate formation in both cell lines. [125I-D-Tyr6,β-Ala11,Phe13, Nle14]Bn-(6-14) bound to both cell lines with high affinity. Neither Bn nor 14 other naturally occurring Bn peptides bound to hBRS-3 with a K(d) 1090 nM. Guanosine 5'-(β,γ- imido)triphosphate inhibited binding to both cells due to a change in receptor affinity. These results demonstrate hBRS-3 has a unique pharmacology. It does not interact with high affinity with any known natural agonist or high affinity antagonist of the Bn receptor family, suggesting the natural ligand is either an undiscovered member of the Bn peptide family or an unrelated peptide. The availability of these cell lines and the hBRS-3 ligand should facilitate identification of the natural ligand for BRS-3, its pharmacology, and cell biology.

    Original languageEnglish (US)
    Pages (from-to)26062-26071
    Number of pages10
    JournalJournal of Biological Chemistry
    Volume272
    Issue number41
    DOIs
    StatePublished - Oct 10 1997

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    Bombesin Receptors
    Bombesin
    Pharmacology
    Cells
    Cell Line
    Ligands
    Peptides
    Norleucine
    Cytology
    3T3 Cells
    Inositol Phosphates
    bombesin receptor subtype 3
    Guanosine
    Non-Small Cell Lung Carcinoma
    Cell Biology
    Availability
    Tissue
    Amino Acids

    ASJC Scopus subject areas

    • Biochemistry

    Cite this

    Discovery of a high affinity radioligand for the human orphan receptor, bombesin receptor subtype 3, which demonstrates that it has a unique pharmacology compared with other mammalian bombesin receptors. / Mantey, Samuel A.; Weber, H. Christian; Sainz, Eduardo; Akeson, Mark; Ryan, Richard R.; Pradhan, Tapas K.; Searles, Robert; Spindel, Eliot; Battey, James F.; Coy, David H.; Jensen, Robert T.

    In: Journal of Biological Chemistry, Vol. 272, No. 41, 10.10.1997, p. 26062-26071.

    Research output: Contribution to journalArticle

    Mantey, Samuel A. ; Weber, H. Christian ; Sainz, Eduardo ; Akeson, Mark ; Ryan, Richard R. ; Pradhan, Tapas K. ; Searles, Robert ; Spindel, Eliot ; Battey, James F. ; Coy, David H. ; Jensen, Robert T. / Discovery of a high affinity radioligand for the human orphan receptor, bombesin receptor subtype 3, which demonstrates that it has a unique pharmacology compared with other mammalian bombesin receptors. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 41. pp. 26062-26071.
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    abstract = "An orphan receptor discovered in 1993 was called bombesin receptor subtype 3 (BRS-3) because of 47-51{\%} amino acid identity with bombesin (Bn) receptors. Its pharmacology is unknown, because no naturally occurring tissues have sufficient receptors to allow studies. We made two cell lines stably expressing the human BRS-3 (hBRS-3). hBRS-3 was overexpressed in the human non-small cell lung cancer cells, NCI-H1299, and the other was made in Balb 3T3 cells, which lack endogenous BRS-3. [D-Phe6,β- Ala11,Phe13,Nle14]Bn-(6-14) (where Nle represents norleucine) was discovered to have high potency for stimulating inositol phosphate formation in both cell lines. [125I-D-Tyr6,β-Ala11,Phe13, Nle14]Bn-(6-14) bound to both cell lines with high affinity. Neither Bn nor 14 other naturally occurring Bn peptides bound to hBRS-3 with a K(d) 1090 nM. Guanosine 5'-(β,γ- imido)triphosphate inhibited binding to both cells due to a change in receptor affinity. These results demonstrate hBRS-3 has a unique pharmacology. It does not interact with high affinity with any known natural agonist or high affinity antagonist of the Bn receptor family, suggesting the natural ligand is either an undiscovered member of the Bn peptide family or an unrelated peptide. The availability of these cell lines and the hBRS-3 ligand should facilitate identification of the natural ligand for BRS-3, its pharmacology, and cell biology.",
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    AU - Weber, H. Christian

    AU - Sainz, Eduardo

    AU - Akeson, Mark

    AU - Ryan, Richard R.

    AU - Pradhan, Tapas K.

    AU - Searles, Robert

    AU - Spindel, Eliot

    AU - Battey, James F.

    AU - Coy, David H.

    AU - Jensen, Robert T.

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