Cloning of cDNAs encoding amphibian bombesin: Evidence for the Relationship between bombesin and gastrin-releasing peptide

Eliot R. Spindel, Bradford W. Gibson, Joseph R. Reeve, Michele Kelly

    Research output: Contribution to journalArticlepeer-review

    23 Scopus citations

    Abstract

    Bombesin is a tetradecapeptide originally isolated from frog skin; its mammalian homologue is the 27-amino acid peptide gastrin-releasing peptide (GRP). cDNAs encoding GRP have been cloned from diverse species, but little is yet known about the amphibian bombesin precursor. Mass spectrometry of HPLC-separated skin exudate from Bombina orientalis was performed to demonstrate the existence of authentic bombesin in the skin of this frog. A cDNA library was prepared from the skin of B. orientalis and mixed oligonucleotide probes were used to isolate cDNAs encoding amphibian bombesin. Sequence analysis revealed that bombesin is encoded in a 119-amino acid prohormone. The carboxyl terminus of bombesin is flanked by two basic amino acids; the amino terminus is not flanked by basic amino acids but is flanked by a chymotryptic-like cleavage site. Northern blot analysis demonstrated similary sized bombesin mRNAs in frog skin, brain, and stomach. Polymerase chain reaction was used to show that the skin and gut bombesin mRNAs encoded the identical prohormones. Prohormone processing, however, differed between skin and gut. Chromatography showed the presence of only authentic bombesin in skin whereas gut extracts contained two peaks of bombesin immunoreactivity, one consistent in size with bombesin and one closer in size to mammalian GRP. Thus the same bombesin prohormone is processed solely to bombesin in skin but is processed to a peptide similar in size to bombesin and to a peptide similar in size to mammalian GRP in stomach.

    Original languageEnglish (US)
    Pages (from-to)9813-9817
    Number of pages5
    JournalProceedings of the National Academy of Sciences of the United States of America
    Volume87
    Issue number24
    DOIs
    StatePublished - 1990

    Keywords

    • Bombina bombina
    • HPLC
    • Mass spectrometry
    • Polymerase chain reaction
    • Prohormone processing

    ASJC Scopus subject areas

    • General

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