Max-Planck-Institut für molekulare Genetik (Abteilung Wittmann), Ihnestrasse 63, D-1000 Berlin (West) 33.
Proc Natl Acad Sci U S A. 1983 May;80(9):2579-83. doi: 10.1073/pnas.80.9.2579.
We describe the theoretical basis of an unconventional method for the determination of the amount of energy transferred between two fluorophores by the Förster mechanism. The method involves an internal comparison made by separation of the fluorophores in situ (i.e., in the optical cell), for example, by means of enzymic digestion; it eliminates several important sources of error and it simplifies calculation while making maximal use of the information contained in the fluorescence spectra. The validity of the method is demonstrated by determination of the known distance between two modifiable sites on the transfer RNA molecule, and its usefulness is exemplified by its application to triangulation of the ribosome of Escherichia coli.
我们描述了一种非传统方法的理论基础，该方法通过福斯特机制来确定两个荧光团之间能量转移的量。该方法涉及通过在原位（即在光学细胞中）分离荧光团进行内部比较，例如通过酶消化；它消除了几个重要的误差源，并简化了计算，同时最大限度地利用了荧光光谱中包含的信息。该方法的有效性通过确定转移 RNA 分子上两个可修饰位点之间的已知距离来证明，并且通过将其应用于大肠杆菌核糖体的三角测量来说明了其有用性。
