Linear-shaped Zero Mode Waveguidesの設計・製作とそれを用いたキネシンおよびATPの1分子観察

カテゴリ: 論文誌(論文単位)

グループ名: 【E】センサ・マイクロマシン部門

発行日: 2017/06/01

タイトル(英語): Design and Fabrication of Linear-shaped Zero Mode Waveguides for Single Molecule Observation of Kinesin and Fluorescent ATP

著者名: 藤本　和也（京都大学大学院工学研究科），森田　有貴（京都大学大学院工学研究科），飯野　亮太（自然科学研究機構 分子科学研究所），富重　道雄（青山学院大学理工学部 物理・数学科），新宅　博文（京都大学大学院工学研究科），小寺　秀俊（京都大学大学院工学研究科），横川　隆司（京都大学大学院工学研究科）

著者名(英語): Kazuya Fujimoto (Kyoto University Graduate School of Engineering), Yuki Morita (Kyoto University Graduate School of Engineering), Ryota Iino (Institute for Molecular Science, National Institute of Natural Sciences), Michio Tomishige (Aoyama Gakuin University, College of Science and Engineering), Hirofumi Shintaku (Kyoto University Graduate School of Engineering), Hidetoshi Kotera (Kyoto University Graduate School of Engineering), Ryuji Yokokawa (Kyoto University Graduate School of Engineering)

キーワード: キネシン，ATP，1分子蛍光観察，ナノトラック，ZMWs，エバネッセント場　　kinesin，ATP，single molecule fluorescent observation，nano-track，ZMWs，evanescent field

要約(英語): Kinesin is a motor protein that transport organelles and proteins by repeating 8 nm discrete steps on a microtubule in eukaryotic cells. The mechanism of kinesin motility coupled with the hydrolysis of adenosine triphosphate (ATP) is of great interest to researchers in biophysics. However, it is still poorly understood when kinesin heads make a step during the ATP hydrolysis and when adenosine diphosphate (ADP) is released from kinesin. In this study, we propose a simultaneous measurement of displacements of kinesin and attachment/detachment of fluorescently-labeled ATP (fATP) using Linear-shaped Zero-Mode Waveguides (LZMWs) that enables to use high concentration of fluorescent molecules for single molecule observation. To this end, we designed LZMWs based on FEM simulation and optimized a fabrication process by electron beam lithography and lift-off process. Assay procedure was established by introducing microtubules into LZMWs utilizing gliding motility. Simultaneous observation in LZMWs revealed that motility of kinesin was preserved and single fATP molecules were visualized under the concentration of 250 nM, which is about ten times higher than in conventional Total Internal Reflection Fluorescence Microscopy (TIRFM).

本誌: 電気学会論文誌E（センサ・マイクロマシン部門誌） Vol.137 No.6 （2017） 特集：第33回「センサ・マイクロマシンと応用システム」シンポジウム受賞論文特集

本誌掲載ページ: 159-164 p

原稿種別: 論文／日本語

電子版へのリンク: https://www.jstage.jst.go.jp/article/ieejsmas/137/6/137_159/_article/-char/ja/