da Graça Miguel M, Eidelman O, Ollivon M, Walter A
Section on Membrane Structure and Function, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Biochemistry. 1989 Oct 31;28(22):8921-8. doi: 10.1021/bi00448a035.
The temperature dependence of octyl glucoside micellization was determined and compared to the phase behavior of the octyl glucoside--egg phosphatidylcholine (PC) mixed system in excess water to help elucidate the process of vesicle formation from mixed surfactant-phospholipid micelles. The critical micelle concentration of octyl glucoside (OG) was determined from the sharp increase of ANS fluorescence at micellization in an NaCl buffer at temperatures ranging from 5 to 40 degrees C. The cmc decreased with increasing temperature from 31 mM at 5 degrees C to 16 mM at 40 degrees C. A similar but less steep temperature dependence is observed for the solubilization of egg PC vesicles by OG as monitored by the surfactant-dependent changes in (1) solution turbidity and (2) the resonance energy transfer between NBD-PE and Rho-PE incorporated in the vesicles. These assays identify two breakpoints, most likely the boundaries of the cylindrical micelle and spheroidal micelle coexistence region. The [OG]aq values at these two breakpoints have similar temperature dependencies. However, the cylindrical mixed micelles at the boundary have nearly identical OG:PC ratios over the temperature range studied, whereas the spheroidal mixed micelles have relatively more OG at the higher temperatures (OG:PC ratio increases from 2.92 to 3.72 between 5 and 35 degrees C). Estimation of the acyl volume to surface area ratio for the compositions observed suggests that this parameter remains constant over temperature. The spheroidal mixed micelles, but not the cylindrical PC-OG micelles, exhibit ideal mixing between the two components at all temperatures (5-35 degrees C). This temperature sensitivity may be utilized to improve the efficacy of membrane protein reconstitution.(ABSTRACT TRUNCATED AT 250 WORDS)
测定了辛基葡糖苷胶束化的温度依赖性,并将其与过量水中辛基葡糖苷 - 卵磷脂(PC)混合体系的相行为进行比较,以帮助阐明由混合表面活性剂 - 磷脂胶束形成囊泡的过程。在5至40摄氏度的温度范围内,通过在NaCl缓冲液中胶束化时ANS荧光的急剧增加来测定辛基葡糖苷(OG)的临界胶束浓度。临界胶束浓度随温度升高而降低,从5摄氏度时的31 mM降至40摄氏度时的16 mM。通过(1)溶液浊度和(2)囊泡中掺入的NBD-PE和Rho-PE之间的共振能量转移所监测的表面活性剂依赖性变化,观察到OG对卵磷脂囊泡增溶的温度依赖性相似但较平缓。这些测定确定了两个断点,最有可能是圆柱形胶束和球形胶束共存区域的边界。这两个断点处的[OG]aq值具有相似的温度依赖性。然而,在所研究的温度范围内,边界处的圆柱形混合胶束具有几乎相同的OG:PC比率,而球形混合胶束在较高温度下具有相对更多的OG(OG:PC比率在5至35摄氏度之间从2.92增加到3.72)。对观察到的组成的酰基体积与表面积比的估计表明,该参数在温度范围内保持恒定。球形混合胶束,而不是圆柱形PC-OG胶束,在所有温度(5 - 35摄氏度)下两种组分之间都表现出理想混合。这种温度敏感性可用于提高膜蛋白重构的效率。(摘要截短于250字)