2011 Fiscal Year Final Research Report

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

Research Project

Project/Area Number	22770153
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Single-year Grants
Research Field	Biophysics
Research Institution	University of Hyogo
Principal Investigator	YAMAMOTO Hitoshi 兵庫県立大学, 大学院・生命理学研究科, 研究員 (40419007)
Project Period (FY)	2010 – 2011
Keywords	1分子計測 / 操作 / 膜タンパク質 / 膜透過
Research Abstract	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.