Single molecular analysis of protein translocation dynamics on the endoplasmic reticulum membrane

蛋白质转运和整合是蛋白质转运到其适当目的地的重要过程。这些过程在真核细胞的内质网膜上进行。在这项研究中，我们的目的是单分子分析的蛋白质转运动力学。利用无细胞蛋白质合成系统和微粒体合成蛋白质转运中间体，并用抑制子-tRNA法进行荧光标记。利用全内反射荧光显微镜成功地观察到了荧光和猝灭。然而，由于高背景光、低荧光强度和短寿命导致的低信噪比，发现几乎观察不到有效的荧光强度变化。因此，提高信噪比仍是今后的工作。

Protein translocation and integration are essential processes by which proteins are transported to their proper destinations. These processes proceed on the endoplasmic reticulum membrane in eukaryotic cells. In this study, we aimed at single molecule analysis of protein translocation dynamics. Protein translocation intermediate were synthesized with using cell free protein synthesis system and microsome, and labeled with fluorescent dye by suppressor-tRNA methods. The fluorescence and quenching were successfully observed by using total internal reflection fluorescence microscopy. However, due to the low S/N ratio caused by high background light, low fluorescence intensity and short life time, it was found that effective fluorescence intensity change was hardly observed. Thus, improving the S/N ratio remained to be as future work.