撤回：使用暗场显微镜对酶促构象变化进行微秒级分辨率研究。

WITHDRAWN: Microsecond resolution of enzymatic conformational changes using dark-field microscopy.

作者信息

Spetzler David, York Justin, Martin James, Ishmukhametov Robert, Frasch Wayne D

机构信息

School of Life Sciences, Arizona State University, P.O. Box 874501, Tempe, AZ 85287-4501, USA.

出版信息

Methods. 2008 Jun 16. doi: 10.1016/j.ymeth.2008.05.004.

DOI:10.1016/j.ymeth.2008.05.004

PMID:18565336

Abstract

We report a novel method to detect angular conformational changes of a molecular motor in a manner sensitive enough to achieve acquisition rates with a time resolution of 2.5mus (equivalent to 400,000fps). We show that this method has sufficient sensitivity to resolve the velocity of the F(1)-ATPase gamma-subunit as it travels from one conformational state to another (transition time). Rotation is detected via a gold nanorod attached to the rotating gamma-subunit of an immobilized F(1)-ATPase. Variations in scattered light intensity allow precise measurement of changes in angular position of the rod below the diffraction limit of light.

摘要

我们报告了一种新颖的方法，用于检测分子马达的角构象变化，其灵敏度足以实现时间分辨率为2.5微秒（相当于400,000帧/秒）的采集速率。我们表明，该方法具有足够的灵敏度来解析F(1)-ATP酶γ亚基从一种构象状态转变为另一种构象状态时的速度（转变时间）。通过附着在固定化F(1)-ATP酶旋转γ亚基上的金纳米棒来检测旋转。散射光强度的变化允许在光的衍射极限以下精确测量棒的角位置变化。