Sakai Norio, Shirai Yasuhito, Saito Naoaki
Department of Molecular and Pharmacological Neuroscience, Graduate School of Biomedical Sciences, Hiroshima University, Japan.
Nihon Yakurigaku Zasshi. 2003 Jun;121(6):421-34. doi: 10.1254/fpj.121.421.
Protein kinase C (PKC) is a family, which consists of at least ten subtypes. To elucidate subtype-specific functions of PKC, we have developed the methods to observe PKC translocation in real time and in the living state using PKC fused with fluorescent proteins, including GFP and DsRed. Based on the live imaging of PKC translocation, we have demonstrated that PKC showed stimulus- and subtype-specific translocation, which can recognize its specific substrate and induce its specific cellular response (PKC targeting). These findings suggest that PKC targeting is the molecular basis underlying the diversity of PKC functions. Live imaging of PKC translocation has been proved to be a beneficial tool for understanding not only PKC functions, but also PKC-mediated signal transduction pathways. We have further analyzed PKC functions in the central nervous system using transgenic mice, which can express PKC-GFP in a brain-region-specific manner.
蛋白激酶C(PKC)是一个家族,由至少十种亚型组成。为了阐明PKC亚型特异性功能,我们开发了利用与荧光蛋白(包括绿色荧光蛋白和红色荧光蛋白)融合的PKC在实时和活细胞状态下观察PKC易位的方法。基于PKC易位的实时成像,我们证明PKC表现出刺激和亚型特异性易位,它能够识别其特定底物并诱导其特定细胞反应(PKC靶向)。这些发现表明PKC靶向是PKC功能多样性的分子基础。PKC易位的实时成像已被证明不仅是理解PKC功能,也是理解PKC介导的信号转导途径的有益工具。我们进一步利用转基因小鼠分析了PKC在中枢神经系统中的功能,这些转基因小鼠能够以脑区特异性方式表达PKC-绿色荧光蛋白。