Laser temperature-jump spectroscopy of intrinsically disordered proteins.
Laser temperature-jump spectroscopy of intrinsically disordered proteins.
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DOI：
10.1007/978-1-4614-3704-8_18
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Hagen, Stephen J
中科院分区：
文献类型：
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作者：
Hagen, Stephen J
激光温度跳跃方法允许实验人员研究非常快速的溶液相过程的动力学和动力学，包括纳秒和微秒时间尺度上的生物分子的构象动力学。结合激光温度跳跃(T-JUMP)激发和适当的光学检测技术(如荧光能量转移)，可以研究蛋白质折叠和结合过程中发生的分子内和分子间构象变化和相互作用。本文介绍了激光温度跳跃方法在研究内在无序蛋白质的偶联折叠和结合的紫外可见荧光研究中的应用。我们强调的实用方面的仪器调整和优化，样品制备，和数据收集使用荧光标记的多肽与紫外激光激发。
Laser temperature-jump methods allow an experimenter to study the kinetics and dynamics of very rapid solution-phase processes, including conformational dynamics of biomolecules on time scales of nanoseconds and microseconds. The combination of laser temperature-jump (T-jump) excitation and appropriate optical detection techniques such as fluorescence energy transfer allows the study of intramolecular and intermolecular conformational changes and interactions that occur during protein folding and binding. This article describes the application of the laser temperature-jump method to UV-visible fluorescence studies of the coupled folding and binding of intrinsically disordered proteins. We emphasize the practical aspects of instrument alignment and optimization, sample preparation, and data collection using fluorescently labeled peptides with UV laser excitation.