Molecular characterization of feline melanocortin 4 receptor and melanocortin 2 receptor accessory protein 2
Molecular characterization of feline melanocortin 4 receptor and melanocortin 2 receptor accessory protein 2
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DOI：
10.1016/j.ygcen.2018.01.020
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发表时间：
2018-05-15
影响因子：
2.7
通讯作者：
Yamamoto, Ichiro
中科院分区：
文献类型：
--
作者：
Habara, Makoto;Mori, Nobuko;Yamamoto, Ichiro
黑素皮质素4受体（Melanocortin 4 receptor, MC4R）是G蛋白偶联受体（GPCR）家族的一员，在中枢神经系统（CNS）中通过α -促黑素细胞激素（α - msh）结合介导能量稳态调节。黑色素皮质素2受体辅助蛋白2 （MRAP2）调节MC4R的功能。我们对猫MC4R和MRAP2进行了cDNA克隆，并对它们的氨基酸序列、猫组织中mRNA的表达模式、蛋白-蛋白相互作用和功能进行了表征。结果表明，与其他哺乳动物MC4R和MRAP2序列同源性较高（>88%），分别编码332和206个氨基酸残基。逆转录-聚合酶链反应分析显示，cat MC4R和MRAP2 mRNA在中枢神经系统中高表达。在转染cat MC4R的CHO-K1细胞中，α - msh刺激以剂量依赖的方式增加细胞内环腺苷一磷酸（cAMP）浓度。此外，MRAP2的存在增强了cat mc4r介导的cAMP产生。这些结果表明，cat MC4R在中枢神经系统中作为神经元介质，其功能受MRAP2调节。此外，我们的NanoBiT研究显示了它们在活细胞中相互作用的动力学；α - msh刺激对MC4R与MRAP2的相互作用有轻微影响，对MC4R的同二聚化没有影响，提示MC4R与MRAP2的相互作用处于基础状态，激活后MC4R的结构变化可能会影响MC4R与MRAP2的相互作用。(C) 2018爱思唯尔公司版权所有。
Melanocortin 4 receptor (MC4R), which is a member of the G protein-coupled receptor (GPCR) family, mediates regulation of energy homeostasis upon the binding of alpha-melanocyte-stimulating hormone (alpha-MSH) in the central nervous system (CNS). Melanocortin 2 receptor accessory protein 2 (MRAP2) modulates the function of MC4R. We performed cDNA cloning of cat MC4R and MRAP2 and characterized their amino acid sequences, mRNA expression patterns in cat tissues, protein-protein interactions, and functions. We found high sequence homology (>88%) with other mammalian MC4R and MRAP2 encoding 332 and 206 amino acid residues, respectively. Reverse transcription-polymerase chain reaction analysis revealed that cat MC4R and MRAP2 mRNA were expressed highly in the CNS. In CHO-K1 cells transfected with cat MC4R, stimulation with alpha-MSH increased intracellular cyclic adenosine monophosphate (cAMP) concentration in a dose-dependent manner. Furthermore, the presence of MRAP2 enhanced the cat MC4R-mediated cAMP production. These results suggested that cat MC4R acts as a neuronal mediator in the CNS and that its function is modulated by MRAP2. In addition, our NanoBiT study showed the dynamics of their interactions in living cells; stimulation with alpha-MSH slightly affected the interaction between MC4R and MRAP2, and did not affect MC4R homodimerization, suggesting that they interact in the basal state and that structural change of MC4R by activation may affect the interaction between MC4R and MRAP2. (C) 2018 Elsevier Inc. All rights reserved.