Cloning of a receptor for amphibian [Phe13]bombesin distinct from the receptor for gastrin-releasing peptide: identification of a fourth bombesin receptor subtype (BB4)
| Autor: | Brenda J. Barry, Peter A. Eden, John T. Taylor, Srinivasa R. Nagalla, Eliot R. Spindel, Kim C. Creswick |
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| Jazyk: | angličtina |
| Rok vydání: | 1995 |
| Předmět: |
Xenopus
Molecular Sequence Data Neuromedin B receptor CHO Cells Biology Transfection digestive system complex mixtures chemistry.chemical_compound Gastrin-releasing peptide Cricetinae Animals Humans Amino Acid Sequence Cloning Molecular Receptor Peptide sequence Phylogeny Mammals Multidisciplinary Sequence Homology Amino Acid Bombesin Brain Neuromedin B Molecular biology Bombesin receptor Rats Receptors Bombesin Kinetics chemistry Gastrin-Releasing Peptide Oocytes Bombesin Receptor Subtype-3 Female Anura Peptides hormones hormone substitutes and hormone antagonists Research Article |
| Popis: | Bombesin is a tetradecapeptide originally isolated from frog skin and demonstrated to have a wide range of actions in mammals. Based on structural homology and similar biological activities, gastrin-releasing peptide (GRP) has been considered the mammalian equivalent of bombesin. We previously reported that frogs have both GRP and bombesin, which therefore are distinct peptides. We now report the cloning of a bombesin receptor subtype (BB4) that has higher affinity for bombesin than GRP. PCR was used to amplify cDNAs related to the known bombesin receptors from frog brain. Sequence analysis of the amplified cDNAs revealed 3 classes of receptor subtypes. Based on amino acid homology, two classes were clearly the amphibian homologs of the GRP and neuromedin B receptors. The third class was unusual and a full-length clone was isolated from a Bombina orientalis brain cDNA library. Expression of the receptor in Xenopus oocytes demonstrated that the receptor responded to picomolar concentrations of [Phe13]-bombesin, the form of bombesin most prevalent in frog brain. The relative rank potency of bombesin-like peptides for this receptor was [Phe13]bombesin > [Leu13]bombesin > GRP > neuromedin B. In contrast, the rank potency for the GRP receptor is GRP > [Leu13]bombesin > [Phe13]bombesin > neuromedin B. Transient expression in CHOP cells gave a Ki for [Phe13]bombesin of 0.2 nM versus a Ki of 2.1 nM for GRP. Distribution analysis showed that this receptor was expressed only in brain, consistent with the distribution of [Phe13]-bombesin. Thus, based on distribution and affinity, this bombesin receptor is the receptor for [Phe13]bombesin. Phylogenetic analysis suggests that this receptor separated prior to separation of the GRP and neuromedin B receptors; thus, BB4 receptors and their cognate ligands may also exist in mammals. |
| Databáze: | OpenAIRE |
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