Two-step FRET as a structural tool.
Two-step FRET as a structural tool.
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DOI：
10.1021/ja034564p
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发表时间：
2003-05
影响因子：
15
通讯作者：
H. Watrob;C. Pan;M. Barkley
中科院分区：
文献类型：
--
作者：
H. Watrob;C. Pan;M. Barkley
FRET研究分子复合物的能力通过使用两个或更多的供体/受体对而扩大。提出了三色团系统中距离测量的一般理论框架。考虑了适用于许多不同情况的三种能量转移方案：(I)在第一和第二发色团之间以及第二和第三发色团之间的两步FRET接力，（II）从单一供体到两个不同受体的FRET，以及（III）在第一和第三发色团之间也有FRET的两步FRET接力。对于供体猝灭和敏化发射测量，推导了涉及多个能量转移步骤的效率方程。该理论得到了已知结构模型系统稳态供体猝灭、寿命猝灭和敏化发射实验数据的支持。在三色团体系中测量的距离与在两色团体系和分子模型中测量的距离一致。最后，讨论了能量传递方案诊断和后续距离测量的标记要求。
The power of FRET to study molecular complexes is expanded by the use of two or more donor/acceptor pairs. A general theoretical framework for distance measurements in three-chromophore systems is presented. Three energy transfer schemes applicable to many diverse situations are considered: (I) two-step FRET relay with FRET between the first and second chromophores and between the second and third, (II) FRET from a single donor to two different acceptors, and (III) two-step FRET relay with FRET also between the first and third chromophores. Equations for the efficiencies involving multiple energy transfer steps are derived for both donor quenching and sensitized emission measurements. The theory is supported by experimental data on model systems of known structure using steady-state donor quenching, lifetime quenching, and sensitized emission. The distances measured in the three-chromophore systems agree with those in two-chromophore systems and molecular models. Finally, labeling requirements for diagnosis of the energy transfer scheme and subsequent distance measurements are discussed.