Khan T K, Chong P L
Department of Biochemistry, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA.
Biophys J. 2000 Mar;78(3):1390-9. doi: 10.1016/S0006-3495(00)76692-6.
Membrane packing and dynamics of bipolar tetraether liposomes composed of the polar lipid fraction E (PLFE) from the thermoacidophilic archaebacterium Sulfolobus acidocaldarius have been studied by perylene fluorescence. At a probe-to-PLFE lipid ratio of 1:400, we have detected an unusual fluorescence intensity increase with increasing temperature, while the fluorescence lifetime changed little. As the ratio was decreased, the intensity anomaly was diminished. At 1:3200 and 1:6400, the anomaly disappeared. A remarkable perylene intensity anomaly was also observed in bilayers composed of saturated monopolar diester phosphatidylcholines at their main phase transition temperatures. These results suggest that the intensity anomaly may be due to probe aggregation caused by tight membrane packing. At the same probe-to-lipid ratio (1:400), however, 1, 2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and 1, 2-diphytanoyl-sn-glycero-3-phosphoglycerol (DPhPG) liposomes did not exhibit any intensity anomaly with increasing temperature. This suggests that DPhPC and DPhPG liposomes are more loosely packed than PLFE liposomes; thus the branched methyl groups are not the contributing factor of the tight membrane packing found in PLFE liposomes. Using a multiexcitation method, we have also determined the average (R), in-plane (R(ip)), and out-of-plane (R(op)) rotational rates of perylene in PLFE liposomes at various temperatures (20-65 degrees C). R and R(ip), determined at two different probe-to-lipid ratios (1:400 and 1:3200), both undergo an abrupt increase when the temperature is elevated to approximately 48 degrees C. These data suggest that PLFE liposomes are rigid and tightly packed at low temperatures, but they begin to possess appreciable "membrane fluidity" at temperatures close to the minimum growth temperature ( approximately 50 degrees C) of thermoacidophilic archaebacteria.
利用苝荧光研究了嗜热嗜酸古细菌嗜酸热硫化叶菌的极性脂质组分E(PLFE)构成的双极四醚脂质体的膜堆积和动力学。在探针与PLFE脂质的比例为1:400时,我们检测到荧光强度随温度升高出现异常增加,而荧光寿命变化不大。随着比例降低,强度异常减弱。在1:3200和1:6400时,异常消失。在饱和单极二酯磷脂酰胆碱构成的双层膜的主相变温度下也观察到显著的苝强度异常。这些结果表明,强度异常可能是由于紧密的膜堆积导致探针聚集所致。然而,在相同的探针与脂质比例(1:400)下,1,2 - 二植烷酰 - sn - 甘油 - 3 - 磷酸胆碱(DPhPC)和1,2 - 二植烷酰 - sn - 甘油 - 3 - 磷酸甘油(DPhPG)脂质体在温度升高时未表现出任何强度异常。这表明DPhPC和DPhPG脂质体的堆积比PLFE脂质体更松散;因此,分支甲基不是PLFE脂质体中紧密膜堆积的促成因素。使用多激发方法,我们还测定了PLFE脂质体中苝在不同温度(20 - 65℃)下的平均旋转速率(R)、面内旋转速率(R(ip))和面外旋转速率(R(op))。在两种不同的探针与脂质比例(1:400和1:3200)下测定的R和R(ip),当温度升高到约48℃时均会突然增加。这些数据表明,PLFE脂质体在低温下是刚性且紧密堆积的,但在接近嗜热嗜酸古细菌的最低生长温度(约50℃)时开始具有明显的“膜流动性”。