A programmable optical angle clamp for rotary molecular motors.
A programmable optical angle clamp for rotary molecular motors.
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DOI：
10.1529/biophysj.106.091074
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发表时间：
2007-07
影响因子：
3.4
通讯作者：
Teuta Pilizota;Thomas Bilyard;F. Bai;M. Futai;Hiroyuki Hosokawa;R. Berry
中科院分区：
文献类型：
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作者：
Teuta Pilizota;Thomas Bilyard;F. Bai;M. Futai;Hiroyuki Hosokawa;R. Berry
光镊在直线分子马达的实验研究中有着广泛的应用。速率和力的依赖性的步骤中的机械化学循环的直线电机进行了探讨，提供详细的了解电机机制。为了研究旋转分子马达，我们设计了一个角度钳光学捕获系统来研究细菌鞭毛马达和F（1）-ATP酶。陷阱位置由数字信号处理板和主机通过声光偏转器控制，电机位置通过三维压电平台控制，电机角度使用一对聚苯乙烯珠作为光陷阱的手柄。使用分辨率高达1度的后焦平面干涉法检测珠对角度，并使用精度高达2度和带宽高达1.6 kHz的反馈算法进行控制。详细的光学陷阱，算法，和对齐程序。初步数据显示的角度控制的F（1）-ATP酶和角度和速度控制的细菌鞭毛马达。
Optical tweezers are widely used for experimental investigation of linear molecular motors. The rates and force dependence of steps in the mechanochemical cycle of linear motors have been probed giving detailed insight into motor mechanisms. With similar goals in mind for rotary molecular motors we present here an optical trapping system designed as an angle clamp to study the bacterial flagellar motor and F(1)-ATPase. The trap position was controlled by a digital signal processing board and a host computer via acousto-optic deflectors, the motor position via a three-dimensional piezoelectric stage and the motor angle using a pair of polystyrene beads as a handle for the optical trap. Bead-pair angles were detected using back focal plane interferometry with a resolution of up to 1 degrees , and controlled using a feedback algorithm with a precision of up to 2 degrees and a bandwidth of up to 1.6 kHz. Details of the optical trap, algorithm, and alignment procedures are given. Preliminary data showing angular control of F(1)-ATPase and angular and speed control of the bacterial flagellar motor are presented.