Isolation of the bioactive peptides CCHamide-1 and CCHamide-2 from Drosophila and their putative role in appetite regulation as ligands for G protein-coupled receptors.
Isolation of the bioactive peptides CCHamide-1 and CCHamide-2 from Drosophila and their putative role in appetite regulation as ligands for G protein-coupled receptors.
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DOI：
10.3389/fendo.2012.00177
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发表时间：
2012
影响因子：
5.2
通讯作者：
Kojima M
中科院分区：
文献类型：
--
作者：
Ida T;Takahashi T;Tominaga H;Sato T;Sano H;Kume K;Ozaki M;Hiraguchi T;Shiotani H;Terajima S;Nakamura Y;Mori K;Yoshida M;Kato J;Murakami N;Miyazato M;Kangawa K;Kojima M
有许多孤儿G蛋白偶联受体(GPCRs)的配体尚未确定。蛙皮素受体亚型3(BRS-3)就是这样一种gpr。BRS-3在糖尿病和肥胖症的发病中起作用。无脊椎动物的GPCR与脊椎动物的GPCR相似。两个果蝇GPCR(CG30106和CG14593)属于BRS-3系统发育亚组。在这里，我们成功地从果蝇全匀浆中利用功能分析和反向药理学技术对果蝇CG30106和CG14593的内源性配体进行了生化纯化，并对其一级氨基酸序列进行了鉴定。纯化的配体被命名为CCHamide-1和CCHamide-2，尽管在结构上与最近从基因组序列搜索中预测的多肽相同。此外，我们的生化特性显示了CCHamide-2的两种N-末端扩展形式。当给苍蝇注射CCHamide-2时，增加了它们的取食动机。我们的结果表明，这些多肽实际上是激活这些受体的主要成分。研究CCHAMIDES的作用将有助于寻找BRS-3配体。
There are many orphan G protein-coupled receptors (GPCRs) for which ligands have not yet been identified. One such GPCR is the bombesin receptor subtype 3 (BRS-3). BRS-3 plays a role in the onset of diabetes and obesity. GPCRs in invertebrates are similar to those in vertebrates. Two Drosophila GPCRs (CG30106 and CG14593) belong to the BRS-3 phylogenetic subgroup. Here, we succeeded to biochemically purify the endogenous ligands of Drosophila CG30106 and CG14593 from whole Drosophila homogenates using functional assays with the reverse pharmacological technique, and identified their primary amino acid sequences. The purified ligands had been termed CCHamide-1 and CCHamide-2, although structurally identical to the peptides recently predicted from the genomic sequence searching. In addition, our biochemical characterization demonstrated two N-terminal extended forms of CCHamide-2. When administered to blowflies, CCHamide-2 increased their feeding motivation. Our results demonstrated these peptides actually present as the major components to activate these receptors in living Drosophila. Studies on the effects of CCHamides will facilitate the search for BRS-3 ligands.