Cloning, expression, and ligand-binding characterization of two neuropeptide Y receptor subtypes in orange-spotted grouper, Epinephelus coioides
Cloning, expression, and ligand-binding characterization of two neuropeptide Y receptor subtypes in orange-spotted grouper, Epinephelus coioides
复制标题
斜带石斑鱼两种神经肽 Y 受体亚型的克隆、表达和配体结合特征
DOI：
10.1007/s10695-014-9960-5
复制
发表时间：
2014-07
影响因子：
2.9
通讯作者：
Wensheng Li
中科院分区：
文献类型：
--
作者：
Fei Wang;Weimin Chen;Haoran Lin;Wensheng Li
神经肽Y （neuropeptide Y， NPY）是一种重要的多肽，它通过不同的亚型受体发挥生理功能。本研究克隆了橙斑石斑鱼（Epinephelus coioides）两种NPY受体（YRs）的全长cdna，命名为npy8br （y8b）和npy2r （y2）。系统发育分析表明，Y8b受体是属于Y1亚家族的硬骨蛋白Y8b受体的同源物，Y2受体是属于Y2亚家族的硬骨蛋白Y2受体的同源物。这两种基因都具有G蛋白偶联受体家族谱。多重比对表明，YRs的细胞外环区域具有不同种类的残基。表达谱分析显示，石斑鱼Y8b受体mRNA主要表达于脑、胃和肠道，而Y2受体mRNA主要表达于脑、卵巢、肝脏和心脏。双免疫荧光分析表明，石斑鱼YRs与石斑鱼NPY在人胚胎肾293T细胞表面周围相互作用。此外，在噬菌体展示系统中进行定点突变，发现Asp（6.59）可能是npy的共同结合位点，而Y8b受体的Asp（2.68）和Y2受体的Glu（5.24）可能参与了亚型特异性结合。结合表达谱和配体结合特征，石斑鱼Y8b受体可能通过脑肠轴参与摄食调节，而石斑鱼Y2受体可能在肝脏和卵巢中平衡Y8b受体的调节活性并参与代谢。
As one of the most important multifunctional peptides, neuropeptide Y (NPY) performs its physiological functions through different subtype receptors. In this study, full-length cDNAs of two NPY receptors (YRs) in orange-spotted grouper (Epinephelus coioides) were cloned and named npy8br (y8b) and npy2r (y2). Phylogenetic analysis indicated that the Y8b receptor is an ortholog of the teleostean Y8b receptor, which belongs to the Y1 subfamily, and the Y2 receptor is an ortholog of the teleostean Y2 receptor, which belongs to the Y2 subfamily. Both of the YRs have G protein-coupled receptor family profiles. Multiple alignments demonstrate that the extracellular loop regions of YRs have distinctive residues of each species. Expression profile analysis revealed that the grouper Y8b receptor mRNA is primarily expressed in the brain, stomach and intestine, while the grouper Y2 receptor mRNA is primarily expressed in the brain, ovary, liver and heart. Double immunofluorescence analysis determined that the grouper YRs interact with the grouper NPY around the human embryonic kidney 293T cell surface. Furthermore, site-directed mutagenesis in a phage display system revealed that Asp(6.59) might be a common NPY-binding site, while Asp(2.68) of the Y8b receptor and Glu(5.24) of the Y2 receptor could be likely involved in subtype-specific binding. Combining the expression profile and ligand-binding feature, the grouper Y8b receptor could be involved in regulating food intake via the brain-gut axis and the grouper Y2 receptor might play a role in balancing the regulatory activity of the Y8b receptor and participate in metabolism in the liver and ovary.
登录
查看更多内容
影响因子：
4.4
作者：
Lindner, Diana;Stichel, Jan;Beck-Sickinger, Annette G.
通讯作者：
Beck-Sickinger, Annette G.
影响因子：
2.7
作者：
Shuge Wu;Bo Li;Hao-ran Lin;Wensheng Li
通讯作者：
Shuge Wu;Bo Li;Hao-ran Lin;Wensheng Li
影响因子：
3.4
作者：
Larsson TA;Olsson F;Sundstrom G;Lundin LG;Brenner S;Venkatesh B;Larhammar D
通讯作者：
Larhammar D
DOI：
10.1017/cbo9780511610561.008
发表时间：
2005
期刊：
--
影响因子：
--
作者：
S. Reed
通讯作者：
S. Reed
影响因子：
3
作者：
Nandkishor R Kotagale;Nikhilesh P. Paliwal;M. M. Aglawe-M.;M. Umekar;B. Taksande
通讯作者：
Nandkishor R Kotagale;Nikhilesh P. Paliwal;M. M. Aglawe-M.;M. Umekar;B. Taksande