CLONING OF A RECEPTOR FOR AMPHIBIAN [PHE(13)]BOMBESIN DISTINCT FROM THE RECEPTOR FOR GASTRIN-RELEASING PEPTIDE - IDENTIFICATION OF A 4TH BOMBESIN RECEPTOR SUBTYPE (BB4)
CLONING OF A RECEPTOR FOR AMPHIBIAN [PHE(13)]BOMBESIN DISTINCT FROM THE RECEPTOR FOR GASTRIN-RELEASING PEPTIDE - IDENTIFICATION OF A 4TH BOMBESIN RECEPTOR SUBTYPE (BB4)
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DOI：
10.1073/pnas.92.13.6205
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发表时间：
1995-06-20
影响因子：
11.1
通讯作者：
SPINDEL, ER
中科院分区：
文献类型：
--
作者：
NAGALLA, SR;BARRY, BJ;SPINDEL, ER
铃蟾肽是一种最初从青蛙皮肤中分离出来的十四肽，并被证明在哺乳动物中具有广泛的作用。基于结构同源性和相似的生物活性，胃泌素释放肽（GRP）被认为是铃蟾肽的哺乳动物等效物。我们之前报道过青蛙同时具有GRP和铃蟾肽，因此它们是不同的肽。现在我们报道了铃蟾肽受体亚型（BB4）的克隆，其对铃蟾肽的亲和力比GRP更高。使用PCR扩增与来自青蛙脑的已知铃蟾肽受体相关的cDNA，对扩增的cDNA进行序列分析揭示了3类受体亚型，基于氨基酸同源性，两类明显是GRP和神经调节素B受体的两栖类同源物，第三类是不寻常的，从侧蚕脑cDNA文库中分离出全长克隆。爪蟾卵母细胞中受体的表达表明，受体对皮摩尔浓度的[Phe(13)]铃蟾肽有反应，铃蟾肽是青蛙脑中最常见的铃蟾肽形式。铃蟾肽样肽对该受体的相对效力等级为[Phe(13)]铃蟾肽>[Leu(13)]铃蟾肽>GRP>神经调节素B。相反，GRP受体的等级效力为GRP> [Leu(13)]铃蟾肽 > [Phe(13)]铃蟾肽 > 神经调节肽 B。CHOP 细胞中的瞬时表达使 [Phe(13)]铃蟾肽的 K-i 为 0.2 nM，而 GRP 的 K-i 为 2.1 nM。分布分析表明，该受体仅在脑中表达，与 [Phe(13)]铃蟾肽的分布一致，因此，根据分布和亲和力，该受体仅在脑中表达。铃蟾肽受体是[Phe(13)]铃蟾肽的受体。系统发育分析表明，该受体先于 GRP 和神经调节素 B 受体分离；因此，BB4受体及其同源配体也可能存在于哺乳动物中。
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 [Phe(13)]bombesin, the form of bombesin most prevalent in frog brain, The relative rank potency of bombesin like peptides for this receptor was [Phe(13)]bombesin > [Leu(13)]bombesin > GRP > neuromedin B. In contrast, the rank potency for the GRP receptor is GRP > [Leu(13)]bombesin > [Phe(13)]bombesin > neuromedin B. Transient expression in CHOP cells gave a K-i for [Phe(13)]bombesin of 0.2 nM versus a K-i of 2.1 nM for GRP, Distribution analysis showed that this receptor was expressed only in brain, consistent with the distribution of [Phe(13)]bombesin, Thus, based on distribution and affinity, this bombesin receptor is the receptor for [Phe(13)]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.