Koushik Srinagesh V, Vogel Steven S
Laboratory of Molecular Physiology, National Institute on Alcohol Abuse and Alcoholism, Bethesda, Maryland 20852, USA.
J Biomed Opt. 2008 May-Jun;13(3):031204. doi: 10.1117/1.2940367.
Forster resonance energy transfer (FRET) is a physical phenomenon used to study molecular interactions in living cells. Changes in the fluorescence lifetime of proteins genetically tagged with a donor fluorophore, such as cyan fluorescent protein or Cerulean, are used to measure energy transfer to a protein tagged with an acceptor fluorophore (yellow fluorescent protein or Venus). Increased transfer efficiency is usually interpreted as closer proximity. Resonance energy transfer is also possible between identical fluorophores. This form of FRET is called energy migration resonance energy transfer (EM-RET). Theoretically, EM-RET should not alter the lifetime or emission spectrum measured from a population of fluorophores. We find a change in the fluorescent lifetime of Cerulean that correlates with energy migration and can result in significant errors when using Cerulean as a donor to measure FRET efficiencies based on fluorescence lifetimes [corrected]
福斯特共振能量转移（FRET）是一种用于研究活细胞中分子相互作用的物理现象。用供体荧光团（如青色荧光蛋白或天蓝蛋白）进行基因标记的蛋白质，其荧光寿命的变化可用于测量向用受体荧光团（黄色荧光蛋白或维纳斯荧光蛋白）标记的蛋白质的能量转移。转移效率的提高通常被解释为距离更近。相同荧光团之间也可能发生共振能量转移。这种形式的FRET称为能量迁移共振能量转移（EM-RET）。理论上，EM-RET不应改变从一群荧光团测得的寿命或发射光谱。我们发现天蓝蛋白的荧光寿命发生了变化，这与能量迁移相关，并且在使用天蓝蛋白作为供体基于荧光寿命测量FRET效率时可能导致显著误差[已校正]
