摘要
目的研究食欲素(orexins,Ox)受体2α与2β能否形成异源二聚体。方法利用激光共聚焦、酶联免疫吸附测定、荧光共振能量转移技术及生物发光能量共振转移技术检测两受体间的相互作用。结果激光共聚焦与酶联免疫吸附测定结果表明,OX2α受体与OX2β受体均在细胞质膜上表达;FRET结果表明,实验组(OX2αR-YFP+OX2βR—CFP)的FRET信号明显强于对照组(YFP+OX2βR—CFP)的FRET信号;BRET结果显示,实验组(OX2αR—YFP+OX2βR-Rluc)的BRET数值(mBRET值为65±15)比对照组(YFP+OX2βR-Rluc/OX2αR—YFP+Rluc)的BRET数值(mBRET值为10±5)高(P〈0.05)。结论OX2αR与OX2βR能够形成组成型的异源二聚体。
Objective To explore the possibility of heterodimerization between orexin type 2α receptor (OX2αR) and orexin type 2β receptor (OX2βR). Methods Using confocal laser scanning microscope, enzyme linked immunosorbent assay ( ELISA), fluorescence resonance energy transfer (FRET) and Bioluminescence resonance energy transfer (BRET) to study the interaction between OX2αR and OX2βR. Result Confocal laser scanning microscope and ELISA showed that OX2αR and OX2βR were both expressed in the cytoplasm.The FRET demonstrated that the signal of the experimental group ( OX2αR-YFP+ OX2βR-CFP) was significantly stronger than that of control group (YFP+OX2βR-CFP).The BRET value of the experimental group (OX2αR-YFP+OX2βR-Pduc, mBRET ratio was 65±15) was higher than that of control group (YFP+ OX2βR-Rluc/OX2αR-YFP+Rluc,mBRET ratio was 10±5 ) (P〈0.05). Conclusion There are hcterodimerization between mOX2αR and mOX2βR.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2016年第4期310-314,共5页
Chinese Journal of Behavioral Medicine and Brain Science
基金
国家自然科学基金(3097108)
山东省高校科技计划项目(J14LK06,JYQ2011KZ001,2012jnjc08,JY2015KJ004)
山东省自然科学基金(ZR2015CL021)
关键词
食欲素受体
荧光共振能量转移
生物发光能量共振转移
二聚体
Orexin receptor
Fluorescence resonance energy transfer
Bioluminescence resonance energy transfer
Heterodimerization