Bagatolli Luis A
Membrane Biophysics and Biophotonics Group/MEMPHYS-Center for Biomembrane Physics, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230, Odense M, Denmark.
Subcell Biochem. 2015;71:105-25. doi: 10.1007/978-3-319-19060-0_5.
A family of polarity sensitive fluorescent probes (2-(dimethylamino)-6-acylnaphtalenes, i.e. LAURDAN, PRODAN, ACDAN) was introduced by Gregorio Weber in 1979, with the aim to monitor solvent relaxation phenomena on protein matrices. In the following years, however, PRODAN and particularly LAURDAN, were used to study membrane lateral structure and associated dynamics. Once incorporated into membranes, the (nanosecond) fluorescent decay of these probes is strongly affected by changes in the local polarity and relaxation dynamics of restricted water molecules existing at the membrane/water interface. For instance, when glycerophospholipid containing membranes undertake a solid ordered (gel) to liquid disordered phase transition the fluorescence emission maximum of these probes shift ~ 50 nm with a significant change in their fluorescence lifetime. Furthermore, the fluorescence parameters of LAURDAN and PRODAN are exquisitely sensitive to cholesterol effects, allowing interpretations that correlate changes in membrane packing with membrane hydration. Different membrane model systems as well as innate biological membranes have been studied with this family of probes allowing interesting comparative studies. This chapter presents a short historical overview about these fluorescent reporters, discusses on different models proposed to explain their sensitivity to membrane hydration, and includes relevant examples from experiments performed in artificial and biological membranes.
1979年,格雷戈里奥·韦伯引入了一类极性敏感荧光探针(2-(二甲基氨基)-6-酰基萘,即劳丹(LAURDAN)、普罗丹(PRODAN)、吖啶丹(ACDAN)),旨在监测蛋白质基质上的溶剂弛豫现象。然而,在随后的几年里,普罗丹,尤其是劳丹,被用于研究膜的侧向结构及相关动力学。一旦掺入膜中,这些探针的(纳秒级)荧光衰减会受到膜/水界面处受限水分子的局部极性和弛豫动力学变化的强烈影响。例如,当含甘油磷脂的膜从固态有序(凝胶)相转变为液态无序相时,这些探针的荧光发射最大值会偏移约50纳米,其荧光寿命也会发生显著变化。此外,劳丹和普罗丹的荧光参数对胆固醇的影响极为敏感,这使得人们能够将膜堆积的变化与膜水合作用联系起来进行解读。利用这类探针研究了不同的膜模型系统以及天然生物膜,从而得以开展有趣的比较研究。本章简要介绍了这些荧光报告分子的历史概况,讨论了为解释它们对膜水合作用的敏感性而提出的不同模型,并列举了在人工膜和生物膜中进行的实验的相关实例。