high-sensitivity real-time imaging of dual protein-protein interactions in living subjects using multicolor luciferases高灵敏度的实时成像双使用多色荧光素酶蛋白质-蛋白质之间的关系生活主题.pdfVIP

High-Sensitivity Real-Time Imaging of Dual Protein- Protein Interactions in Living Subjects Using Multicolor Luciferases 1 2 1 3 1 3 Naoki Hida , Muhammad Awais , Masaki Takeuchi , Naoto Ueno , Mayuri Tashiro , Chiyo Takagi , 3 3 4,5 1,6 Tanuja Singh , Makoto Hayashi , Yoshihiro Ohmiya , Takeaki Ozawa * 1 Department of Chemistry, Graduate School of Science, The University of Tokyo, Bunkyo-Ku, Tokyo, Japan, 2 Centre for Cell Imaging, School of Biological Science, The University of Liverpool, Liverpool, United Kingdom, 3 National Institute for Basic Biology, National Institutes of Natural Sciences, Okazaki, Aichi, Japan, 4 Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Osaka, Japan, 5 Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, Japan, 6 PRESTO, Japan Science and Technology Agency, Tokyo, Japan Abstract Networks of protein-protein interactions play key roles in numerous important biological processes in living subjects. An effective methodology to assess protein-protein interactions in living cells of interest is protein-fragment complement assay (PCA). Particularly the assays using fluorescent proteins are powerful techniques, but they do not directly track interactions because of its irreversibility or the time for chromophore formation. By contrast, PCAs using bioluminescent proteins can overcome these drawbacks. We herein describe an imaging method for real-time analysis of protein-protein interactions using multicolor luciferases with different spectral characteristics. The sensitivity and signal-to-background