| 姜云璐,田艳君,王正文,吕志瞳,白波,陈京.FRET技术研究Apelin受体与G蛋白的相互作用[J].济宁医学院学报,2017,40(1):9-15 |
| FRET技术研究Apelin受体与G蛋白的相互作用 |
| Using fluorescence resonance energy transfer technology to study the interaction between G protein and Apelin receptor |
| 投稿时间：2016-09-21 |
| DOI：10.3969/j.issn.1000-9760.2017.01.003 |
| 中文关键词: Apelin受体;荧光共振能量转移技术;信号转导;G蛋白偶联受体 |
| 英文关键词: Apelin receptor;Fluorescence resonance energy transfer technology;Signal transduction;G-protein-coupled receptors |
| 基金项目:国家自然基金项目（3097108；816712276；31271243；81070961）；山东省高校科技计划项目（ZR2014HL404；JY2015KJ004；JY2015KJ002；2015-57-6；2015-57-62；J15LE19） |
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| 中文摘要: |
| 目的 探讨Apelin受体与G蛋白之间的相互作用。方法 在HEK293T细胞中建立Apelin受体与各亚型G蛋白的过表达体系，利用荧光共振能量转移技术（fluorescence resonance energy transfer technology，FRET）实时、动态、连续的监测Apelin受体与G蛋白偶联的机制。结果 加入激动剂 Apelin-13刺激后，Apelin受体与Gαi1亚基的FRET值没有发生明显的变化（P> 0.05），Apelin受体与Gαi2、Gαi3亚基的FRET信号显著增加（P< 0.05），而Apelin受体与Gαo、Gαq亚基的FRET信号相对于刺激前显著降低（P< 0.05）。结论 Apelin受体在其激动剂Apelin-13作用下，能够激活G蛋白的Gαi2、Gαi3、Gαo、Gαq亚基，并引起Gαi2、Gαi3亚基构象重排进而与Gγ亚基相互靠近，Gαo、Gαq亚基远离Gγ2亚基，而不与Gαi1亚基发生相互作用。 |
| 英文摘要: |
| Objective To explore the interaction between Apelin receptor and G protein.Methods To monitor Apelin receptor mediated activation of G proteins in living cells in real-time and dynamic,and the established cell lines that expressed APJ or some one of G protein using fluorescence resonance energy transfer technology.Results Upon superfusion with Apelin-13 and recording of whole-cell uorescence,there was no significant change in FRET ratio (P> 0.05).There was an agonist-induced increase in FRET in in HEK293T cells which transfected with Gαi2 /Gαi3,Gα1Gα2-CFP and Apelin receptor (P< 0.05).And there was an agonist-induced decrease in FRET in Gαo/Gαq with APJ(P< 0.05).Conclusion These results demonstrate that activation of APJ with Apelin-13 also leads to the activation of Gαi,Gαo,Gαq proteins,and Apelin receptor activation causes Gαi2 and Gαi3 subunits activation through a unique rearrangement process instead of subunit dissociation,and leads to Gαo and Gαq disassociation with Gα1α2 and Gαi1 is not activated following APJ activity or APJ only cause a very small portion of the Gαi1 activation. |
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