Hattori Mitsuru, Ozawa Takeaki
Department of Chemistry, School of Science, The University of Tokyo.
Anal Sci. 2015;31(4):327-30. doi: 10.2116/analsci.31.327.
For the design of therapeutic drugs, G protein coupled receptors (GPCRs) are notable targets. Many screening methods have been developed to identify effective agents for GPCR signaling. However, analyses of temporal variations of GPCR activity with specific ligands remain insufficient because of monitoring method limitations and difficulties. We previously developed a high-throughput bioluminescence measuring system to detect interactions of GPCR with β-arrestin based on split luciferase fragment complementation. By newly introducing a bioluminescence imaging technique into the system, we demonstrate a method for the temporal monitoring of GPCR-β-arrestin interactions in living cells during stimulation by different ligands.
对于治疗药物的设计而言，G蛋白偶联受体（GPCRs）是值得关注的靶点。已经开发了许多筛选方法来鉴定用于GPCR信号传导的有效药物。然而，由于监测方法的局限性和困难，对特定配体存在时GPCR活性的时间变化分析仍然不足。我们之前开发了一种高通量生物发光测量系统，基于分裂荧光素酶片段互补来检测GPCR与β-抑制蛋白的相互作用。通过在该系统中新引入生物发光成像技术，我们展示了一种在不同配体刺激期间对活细胞中GPCR-β-抑制蛋白相互作用进行时间监测的方法。
