Antunes-Madeira M C, Almeida L M, Madeira V M
Centro de Biologia Celular, Departamento de Zoologia, Coimbra, Portugal.
Biochim Biophys Acta. 1990 Feb 16;1022(1):110-4. doi: 10.1016/0005-2736(90)90406-e.
Fluorescence polarization studies of 1,6-diphenyl-1,3,5-hexatriene (DPH) have been compared with the excimer/monomer fluorescence intensity ratio (I'/I) of 1,3-di(2-pyrenyl)propane, (2Py(3)2Py). This ratio permits evaluation of changes in fluidity of the outer regions of the bilayer, where 2Py(3)2Py preferentially distributes. On the other hand, fluorescence polarization of DPH reports the structural order of the bilayer core. In the fluid phase of DMPC bilayers, for lindane concentrations higher than 25 microM, the excimer/monomer fluorescence intensity ratio (I'/I) decreases, thus reflecting an order increase of the probe environment. However, in the same conditions, the fluorescence polarization of DPH is almost insensitive to any perturbation. Identical results have been obtained in other pure lipid bilayers, namely DPPC and DSPC. However, both probes detect disordering effects of lindane in the gel phase of these lipids. The pyrene probe, unlike DPH, is very sensitive to the pretransitions of DPPC and DSPC, removed in the presence of lindane. Both probes fail to detect any apparent effect of lindane in DMPC bilayers enriched with high cholesterol content (greater than 30 mol%). However, in DMPC bilayers with low cholesterol content (less than 30 mol%), for temperatures below the phase transition of DMPC, both probes detect fluidizing effects induced by lindane. Nevertheless, above the phase transition of DMPC, 2Py(3)2Py detects ordering effects of lindane, whereas DPH detects hardly any effect. These results in DMPC bilayers with low cholesterol content are qualitatively similar to those described for DMPC without cholesterol.
已将1,6 - 二苯基 - 1,3,5 - 己三烯(DPH)的荧光偏振研究与1,3 - 二(2 - 芘基)丙烷(2Py(3)2Py)的准分子/单体荧光强度比(I'/I)进行了比较。该比值可用于评估双层膜外层区域流动性的变化,2Py(3)2Py优先分布于此区域。另一方面,DPH的荧光偏振反映了双层膜核心的结构有序性。在二肉豆蔻酰磷脂酰胆碱(DMPC)双层膜的流体相中,对于林丹浓度高于25 microM的情况,准分子/单体荧光强度比(I'/I)降低,从而反映出探针环境的有序性增加。然而,在相同条件下,DPH的荧光偏振对任何扰动几乎都不敏感。在其他纯脂质双层膜,即二棕榈酰磷脂酰胆碱(DPPC)和二硬脂酰磷脂酰胆碱(DSPC)中也得到了相同的结果。然而,两种探针都检测到林丹在这些脂质凝胶相中的无序化作用。与DPH不同,芘探针对DPPC和DSPC的预转变非常敏感,在林丹存在下这种预转变会消失。两种探针都未能检测到林丹在富含高胆固醇含量(大于30摩尔%)的DMPC双层膜中的任何明显作用。然而,在低胆固醇含量(小于30摩尔%)的DMPC双层膜中,对于低于DMPC相变温度的情况,两种探针都检测到了林丹诱导的流化作用。然而,在DMPC相变温度以上,2Py(3)2Py检测到林丹的有序化作用,而DPH几乎检测不到任何作用。在低胆固醇含量的DMPC双层膜中的这些结果在定性上与不含胆固醇的DMPC所描述的结果相似。