Purification and characterization of bioactive peptides RYamide and CCHamide in the kuruma shrimp Marsupenaeus japonicus
Purification and characterization of bioactive peptides RYamide and CCHamide in the kuruma shrimp Marsupenaeus japonicus
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DOI：
10.1016/j.ygcen.2017.01.008
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发表时间：
2017-05-15
影响因子：
2.7
通讯作者：
Ida, Takanori
中科院分区：
文献类型：
--
作者：
Mekata, Tohru;Kono, Tomoya;Ida, Takanori
为了了解食欲调节系统，使用表达果蝇 G 蛋白偶联受体 (GPCR) CG5811（一种 RYamide 受体）或 CG14593（一种 CCHamide-2 受体）的 CHO 细胞，通过反向药理学测定分离和纯化来自车虾 Marsupenaeus japonicus (Mj) 的生物活性肽。获得了四种具有GPCR结合活性的肽，并将其命名为Mj RYamide-1、Mj RYamide-2、Mj RYamide-3和Mj CCHamide。使用库鲁玛虾转录组数据库鉴定了编码这些肽的前原肽的基因。 Mj prepro-RYamide 基因编码含有 Mj RYamide-1、Mj RYamide-2 和 Mj RYamide-3 的 130 个氨基酸的多肽，而 Mj prepro-CCHamide 基因编码含有单个 Mj CCHamide 肽的 119 个氨基酸的多肽。这些基因的表达在包括大脑和腹神经索在内的各种神经器官中得到证实。此外，饥饿后大脑中前原-RYamide 基因表达显着降低。因此，RYamides 可能与摄食或消化的调节有关。还研究了施用合成肽后皮艇（无脊椎动物中的 c-fos 直系同源物）基因表达的变化。用 Mj RYamide-3 或 CCHamide 处理后，肝胰腺中的 Mj kayak 表达水平上调。因此，本研究中分离的肽可能对水产养殖无脊椎动物的细胞代谢具有一定的调节作用。 (C) 2017 年，爱思唯尔公司出版
To understand the regulation systems of appetite, bioactive peptides from the kuruma shrimp Marsupenaeus japonicus (Mj) were isolated and purified by reverse pharmacological assays using CHO cells expressing the Drosophila melanogaster G-protein-coupled receptors (GPCRs) CG5811 (a RYamide receptor) or CG14593 (a CCHamide-2 receptor). Four peptides having binding activity to GPCRs were obtained and named Mj RYamide-1, Mj RYamide-2, Mj RYamide-3, and Mj CCHamide. Genes encoding the prepropeptides of these peptides were identified using kuruma shrimp transcriptome databases. The Mj prepro-RYamide gene encodes a 130-amino acid polypeptide containing Mj RYamide-1, Mj RYamide-2, and Mj RYamide-3, whereas the Mj prepro-CCHamide gene encodes a 119-amino acid polypeptide containing a single Mj CCHamide peptide. The expression of these genes was confirmed in various neuronal organs including the brain and ventral nerve cord. In addition, prepro-RYamide gene expression is significantly reduced in the brain after starvation. RYamides may thus be associated with regulation of feeding or digestion. Changes in kayak (the c-fos ortholog in invertebrates) gene expression after administration of synthetic peptides were also investigated. Mj kayak expression levels are upregulated in hepatopancreas after treatment with Mj RYamide-3 or CCHamide. Thus, the peptides isolated in this study may have some regulatory effect on cellular metabolism in aquacultured invertebrates. (C) 2017 Published by Elsevier Inc.