期刊
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
卷 297, 期 2, 页码 C290-C298出版社
AMER PHYSIOLOGICAL SOC
DOI: 10.1152/ajpcell.00647.2008
关键词
cochlear amplifier; outer hair cell; conformational change; total internal reflection fluorescence
资金
- Ministry of Education, Science, Sports, Culture
- National Science Foundation
- Japan Society for Promotion of Science
Gleitsman KR, Tateyama M, Kubo Y. Structural rearrangements of the motor protein prestin revealed by fluorescence resonance energy transfer. Am J Physiol Cell Physiol 297: C290-C298, 2009. First published June 10, 2009; doi: 10.1152/ajpcell.00647.2008. Prestin is a membrane protein expressed in the outer hair cells (OHCs) in the cochlea that is essential for hearing. This unique motor protein transduces a change in membrane potential into a considerable mechanical force, which leads to a cell length change in the OHC. The nonlinear capacitance in cells expressing prestin is recognized to reflect the voltage-dependent conformational change of prestin, of which its precise nature remains unknown. In the present work, we aimed to detect the conformational changes of prestin by a fluorescence resonance energy transfer (FRET)-based technique. We heterologously expressed prestin labeled with fluorophores at the COOH-or NH2-terminus in human embryonic kidney-293T cells, and monitored FRET changes on depolarization-inducing high KCl application. We detected a significant decrease in intersubunit FRET both between the COOH-termini and between the COOH-and NH2-termini. A similar FRET decrease was observed when membrane potential was directly and precisely controlled by simultaneous patch clamp. Changes in FRET were suppressed by either of two treatments known to abolish nonlinear capacitance, V499G/Y501H mutation and sodium salicylate. Our results are consistent with significant movements in the COOH-terminal domain of prestin upon change in membrane potential, providing the first dynamic information on its molecular rearrangements.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据