Fluorescence quenching by TEMPO:: A sub-30 Å single-molecule ruler
Fluorescence quenching by TEMPO:: A sub-30 Å single-molecule ruler
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DOI：
10.1529/biophysj.105.071027
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发表时间：
2005-11-01
影响因子：
3.4
通讯作者：
Deniz, AA
中科院分区：
文献类型：
--
作者：
Zhu, PZ;Clamme, JP;Deniz, AA
合成了一系列以5-羧基四甲基罗丹明（5-TAMRA）和小氮氧自由基TEMPO标记的DNA分子，并测试了分子内猝灭效率是否可以用于小集合和单分子实验中短分子内距离的测量。结合分子力学模型的距离计算，稳态系综荧光和荧光相关光谱测量的实验结果都表明，在10-30埃范围内，猝灭速率常数随染料猝灭剂距离的增加呈指数下降。结果表明，TEMPO-5-TAMRA。荧光猝灭是一种很有前途的测量单个生物分子短距离变化的方法。
A series of DNA molecules labeled with 5-carboxytetramethylrhodamine (5-TAMRA) and the small nitroxide radical TEMPO were synthesized and tested to investigate whether the intramolecular quenching efficiency can be used to measure short intramolecular distances in small ensemble and single-molecule experiments. In combination with distance calculations using molecular mechanics modeling, the experimental results from steady-state ensemble fluorescence and fluorescence correlation spectroscopy measurements both show an exponential decrease in the quenching rate constant with the dye-quencher distance in the 10-30 angstrom range. The results demonstrate that TEMPO-5-TAMRA. fluorescence quenching is a promising method to measure short distance changes within single biomolecules.