Proks P, Hianik T, Kvasnicka P
Institute of Molecular Physiology and Genetics, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Gen Physiol Biophys. 1992 Oct;11(5):441-58.
The effects of fluorescent probes 9-aminoacridine (9AA) and atebrine (AT) on physical properties of liposomes and planar bilayer lipid membranes (BLM) were studied. The method of fluorescence spectroscopy and the electrostriction method based on measurement of higher current harmonics were used. At low concentrations (10(-5)-5 x 10(-5) mol/l), 9AA increased fluorescence intensity, while in liposomes from soybean phosphatidylcholine fluorescence quenching occurred at higher probe concentration. Fluorescence quenching occurred over the entire concentration range tested (10(-5)-10(-4) mol/l) in liposomes made from a mixture of egg phosphatidylcholine and cardiolipin. In contrast to 9AA, AT, thanks to its hydrophobic chain, penetrates deeper into the hydrophobic membrane moiety; thus, immobilization of the molecule and an increase in fluorescence intensity was always observed. Probes adsorbed to membranes, leaving their electric capacitance effectively unchanged. Adsorption of charged dye particles induced small changes in transmembrane potential. In the presence of 10(-5) mol/l AT, the modulus of elasticity E perpendicular increased somewhat for soft membranes (E perpendicular approximately 2.5 x 10(7) Pa), whereas it decreased for hard membranes (E perpendicular approximately 5 x 10(7) Pa). pH gradient present on the membrane affected the ability of the dyes to incorporate into the membranes. Our results provide evidence against the proposed model of the quenching mechanism introduced by Rottenberg and Lee (1975).
研究了荧光探针9-氨基吖啶(9AA)和阿的平(AT)对脂质体和平面双层脂质膜(BLM)物理性质的影响。采用了荧光光谱法和基于测量高次电流谐波的电致伸缩法。在低浓度(10^(-5)-5×10^(-5)mol/L)时,9AA增加荧光强度,而在大豆磷脂酰胆碱脂质体中,较高探针浓度时发生荧光猝灭。在由鸡蛋磷脂酰胆碱和心磷脂混合物制成的脂质体中,在整个测试浓度范围(10^(-5)-10^(-4)mol/L)内均发生荧光猝灭。与9AA不同,AT由于其疏水链,能更深入地渗透到疏水膜部分;因此,总是观察到分子的固定化和荧光强度的增加。探针吸附到膜上,其电容实际上保持不变。带电染料颗粒的吸附引起跨膜电位的微小变化。在存在10^(-5)mol/L AT的情况下,软膜(E⊥约为2.5×10^7Pa)的垂直弹性模量E有所增加,而硬膜(E⊥约为5×10^7Pa)的垂直弹性模量E则降低。膜上存在的pH梯度影响染料掺入膜的能力。我们的结果为反对Rottenberg和Lee(1975)提出的猝灭机制模型提供了证据。