The essential role for aromatic cluster in the beta 3 adrenergic receptor
文献类型：期刊论文
| 作者 | Cai, Hai-yan; Xu, Zhi-jian ; Tang, Jie; Sun, Ying; Chen, Kai-xian; Wang, He-yao; Zhu, Wei-liang
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| 刊名 | ACTA PHARMACOLOGICA SINICA
 |
| 出版日期 | 2012-08 |
| 卷号 | 33期号:8页码:1062-1068 |
| 关键词 | beta 3 adrenergic receptor constitutive activity mutation aromatic residue G protein-coupled receptors |
| ISSN号 | 1671-4083 |
| DOI | 10.1038/aps.2012.55 |
| 文献子类 | Article |
| 英文摘要 | Aim: To explore the function of the conserved aromatic cluster F213(5.47), F308(6.51), and F309(6.52) in human beta 3 adrenergic receptor (h beta 3AR). Methods: Point mutation technology was used to produce plasmid mutations of h beta 3AR. HEK-293 cells were transiently co-transfected with the h beta 3AR (wild-type or mutant) plasmids and luciferase reporter vector pCRE-luc. The expression levels of h beta 3AR in the cells were determined by Western blot analysis. The constitutive signalling and the signalling induced by the beta 3AR selective agonist, BRL (BRL37344), were then evaluated. To further explore the interaction mechanism between BRL and beta 3AR, a three-dimensional complex model of beta 3AR and BRL was constructed by homology modelling and molecular docking. Results: For F308(6.51), Ala and Leu substitution significantly decreased the constitutive activities of beta 3AR to approximately 10% of that for the wild-type receptor. However, both the potency and maximal efficacy were unchanged by Ala substitution. In the F308(6.51)L construct, the EC50 value manifested as a "right shift" of approximately two orders of magnitude with an increased E-max. Impressively, the molecular pharmacological phenotype was similar to the wild-type receptor for the introduction of Tyr at position 308(6.51), though the EC50 value increased by approximately five-fold for the mutant. For F309(6.52), the constitutive signalling for both F309(6.52)A and F309(6.52)L constructs were strongly impaired. In the F309(6.52)A construct, BRL-stimulated signalling showed a normal E-max but reduced potency. Leu substitution of F309(6.52) reduced both the E-max and potency. When F309(6.52) was mutated to Tyr, the constitutive activity was decreased approximately three-fold, and BRL-stimulated signalling was significantly impaired. Furthermore, the double mutant (F308(6.51)A_F309(6.52)A) caused the total loss of beta 3AR function. The predicted binding mode between beta 3AR and BRL revealed that both F308(6.51) and F309(6.52) were in the BRL binding pocket of beta 3AR, while F213(5.47) and W305(6.48) were distant from the binding site. Conclusion: These results revealed that aromatic residues, especially F308(6.51) and F309(6.52), play essential roles in the function of beta 3AR. Aromatic residues maintained the receptor in a partially activated state and significantly contributed to ligand binding. The results supported the common hypothesis that the aromatic cluster F[Y]5.47/F[Y]6.52/F[Y]6.51 conserved in class A G protein-coupled receptor (GPCR) plays an important role in the structural stability and activation of GPCRs. |
| WOS关键词 | PROTEIN-COUPLED RECEPTOR ; ANGSTROM CRYSTAL-STRUCTURE ; TOGGLE SWITCH MODEL ; CONFORMATIONAL-CHANGES ; BETA(2) ADRENOCEPTOR ; FUNCTIONAL-ROLE ; ACTIVATION ; RHODOPSIN ; LOCK ; MECHANISMS |
| 资助项目 | National Natural Science Foundation of China[20721003] ; National Natural Science Foundation of China[81072681] ; International ST Cooperation[2010DFB73280] ; Shanghai Committee of Science and Technology International Cooperation Project[09540703900] |
| WOS研究方向 | Chemistry ; Pharmacology & Pharmacy |
| 语种 | 英语 |
| CSCD记录号 | CSCD:4602927 |
| WOS记录号 | WOS:000307233900012 |
| 出版者 | ACTA PHARMACOLOGICA SINICA |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/278001]  |
| 专题 | 药理学第三研究室 药物发现与设计中心 |
| 通讯作者 | Wang, He-yao |
| 作者单位 | Chinese Acad Sci, Shanghai Inst Mat Med, Drug Discovery & Design Ctr, Shanghai 201203, Peoples R China |
| 推荐引用方式 GB/T 7714 | Cai, Hai-yan,Xu, Zhi-jian,Tang, Jie,et al. The essential role for aromatic cluster in the beta 3 adrenergic receptor[J]. ACTA PHARMACOLOGICA SINICA,2012,33(8):1062-1068. |
| APA | Cai, Hai-yan.,Xu, Zhi-jian.,Tang, Jie.,Sun, Ying.,Chen, Kai-xian.,...&Zhu, Wei-liang.(2012).The essential role for aromatic cluster in the beta 3 adrenergic receptor.ACTA PHARMACOLOGICA SINICA,33(8),1062-1068. |
| MLA | Cai, Hai-yan,et al."The essential role for aromatic cluster in the beta 3 adrenergic receptor".ACTA PHARMACOLOGICA SINICA 33.8(2012):1062-1068. |
入库方式： OAI收割
来源：上海药物研究所
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