Peng K, Visser A J, van Hoek A, Wolfs C J, Sanders J C, Hemminga M A
Department of Molecular Physics, Agricultural University, Wageningen, The Netherlands.
Eur Biophys J. 1990;18(5):277-83. doi: 10.1007/BF00188040.
Fluorescent probes located in heterogeneous environments give rise to anomalous time-resolved fluorescence anisotropy. A simple analytical expression of anisotropy has been derived for the case of a small difference in local fluorescence lifetimes. The expression has the diagnostic advantage that the time dependence of the fluorescence anisotropy can be predicted from the differences in fluorescence lifetimes and residual anisotropies of the probes located in different sites. Using this model, the local fluorescence anisotropy parameters and the relative contributions of the lipid probe octadecyl rhodamine B in a lipid environment and in the vicinity of bacteriophage M13 coat protein reconstituted in phospholipid bilayers, composed of 80% 1,2-dimyristoyl-sn-glycero-3-phosphocholine and 20% 1,2-dimyristoyl-sn-glycero-3-phosphoglycerol have been determined experimentally. At 40 degrees C, the correlation times for bound and free probes are 2.3 and 3.0 ns, respectively, while the corresponding order parameters are 0.85 and 0.62, respectively.
位于异质环境中的荧光探针会产生异常的时间分辨荧光各向异性。对于局部荧光寿命存在微小差异的情况，已经推导出了一个简单的各向异性分析表达式。该表达式具有诊断优势，即可以根据位于不同位点的探针的荧光寿命差异和残余各向异性来预测荧光各向异性的时间依赖性。使用该模型，通过实验确定了脂质探针十八烷基罗丹明B在脂质环境中以及在由80% 1,2 - 二肉豆蔻酰 - sn - 甘油 - 3 - 磷酸胆碱和20% 1,2 - 二肉豆蔻酰 - sn - 甘油 - 3 - 磷酸甘油组成的磷脂双层中重构的噬菌体M13外壳蛋白附近的局部荧光各向异性参数和相对贡献。在40摄氏度时，结合态和游离态探针的相关时间分别为2.3纳秒和3.0纳秒，而相应的序参数分别为0.85和0.62。
