Time-resolved methods in biophysics. 9. Laser temperature-jump methods for investigating biomolecular dynamics
Time-resolved methods in biophysics. 9. Laser temperature-jump methods for investigating biomolecular dynamics
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DOI：
10.1039/b819929a
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发表时间：
2009-01-01
影响因子：
3.1
通讯作者：
Kubelka, Jan
中科院分区：
文献类型：
--
作者：
Kubelka, Jan
许多重要的生化过程发生在纳秒和微秒的时间尺度上。十多年前，激光温度跳跃(T-JUMP)被引入生物物理学，为直接的实验观测打开了这些以前无法获得的时间区域。从那时起，激光T跳跃方法已经发展成为研究快速生物分子动力学的最通用和最通用的方法之一。本文综述了激光T跳跃技术的原理及其在生物物理学中的应用。简要介绍了T跳跃的驰豫动力学和激光T跳跃方法的历史发展。总结了激光T跳跃实验设计中重要的物理原理和实际实验考虑。这些包括用于产生加热脉冲的拉曼转换，对温度跳跃的大小、持续时间和一致性的考虑，以及光声波、空化和热透镜的潜在不利影响及其消除。详细介绍了美国国立卫生研究院化学物理实验室研制的激光T-JUMP装置，并对目前使用的其他激光T-JUMP设计作了简要介绍。最后，综述了激光T跳跃在生物物理学中的应用，重点介绍了激光T跳跃方法在生物分子过程动力学中提供了重要的新结果和见解的广泛问题。
Many important biochemical processes occur on the time-scales of nanoseconds and microseconds. The introduction of the laser temperature-jump (T-jump) to biophysics more than a decade ago opened these previously inaccessible time regimes up to direct experimental observation. Since then, laser T-jump methodology has evolved into one of the most versatile and generally applicable methods for studying fast biomolecular kinetics. This perspective is a review of the principles and applications of the laser T-jump technique in biophysics. A brief overview of the T-jump relaxation kinetics and the historical development of laser T-jump methodology is presented. The physical principles and practical experimental considerations that are important for the design of the laser T-jump experiments are summarized. These include the Raman conversion for generating heating pulses, considerations of size, duration and uniformity of the temperature jump, as well as potential adverse effects due to photo-acoustic waves, cavitation and thermal lensing, and their elimination. The laser T-jump apparatus developed at the NIH Laboratory of Chemical Physics is described in detail along with a brief survey of other laser T-jump designs in use today. Finally, applications of the laser T-jump in biophysics are reviewed, with an emphasis on the broad range of problems where the laser T-jump methodology has provided important new results and insights into the dynamics of the biomolecular processes.