Pühse Matthias, Jeworrek Christoph, Winter Roland
University of Dortmund, Department of Chemistry, Physical Chemistry I - Biophysical Chemistry, Otto-Hahn-Strasse 6, D-44227 Dortmund, Germany.
Chem Phys Lipids. 2008 Mar;152(1):57-63. doi: 10.1016/j.chemphyslip.2008.01.002. Epub 2008 Jan 11.
We investigated the phase behaviour of the binary lipid mixture dipalmitoylphosphatidylcholine (DPPC)-ergosterol (78:22mol%) as a function of temperature and pressure using Fourier transform-infrared (FT-IR) spectroscopy and small-angle X-ray scattering (SAXS). A temperature range from 10 to 70 degrees C and a pressure range from 1 to approximately 8500bar was covered to be able to establish a p,T-phase diagram of this model mixture of a fungal plasma membrane. Our results show a liquid-ordered/solid-ordered (l(o)+s(o)) two-phase coexistence region up to 41 degrees C, followed by a liquid-disordered/liquid-ordered (l(d)+l(o)) coexistence region up to approximately 57.5 degrees C, giving way to an all-fluid-like l(d) phase at higher temperatures. With increasing pressure, all phase boundaries shift upwards to higher temperatures. The width of the raft-like l(d)+l(o) two-phase coexistence region is found to be about 2kbar over the temperature range covered. At higher pressures, transitions to a l(o)+s(o) two-phase region and, at still higher pressures and low temperatures, an all-ordered s(o) phase is detected. Finally, the biological (high pressure-induced upregulation of ergosterol synthesis) and biotechnological (inactivation of fungal microorganisms) relevance of these studies is discussed. The results may be particularly interesting for understanding the function of membrane proteins under external stress conditions, such as high pressure. Interestingly, in this pressure range of about 2kbar, ceasing of membrane protein function has been observed in natural membrane environments. They may be inactivated under pressure/temperature conditions when the plasma membrane becomes too rigid, i.e., reaches an overall ordered conformational state.
我们使用傅里叶变换红外(FT - IR)光谱和小角X射线散射(SAXS)研究了二棕榈酰磷脂酰胆碱(DPPC)-麦角固醇(78:22摩尔%)二元脂质混合物的相行为随温度和压力的变化。覆盖了10至70摄氏度的温度范围和1至约8500巴的压力范围,以便能够建立这种真菌质膜模型混合物的p,T相图。我们的结果表明,在高达41摄氏度时存在液序/固序(l(o)+s(o))两相共存区域,随后在高达约57.5摄氏度时出现液无序/液序(l(d)+l(o))共存区域,在更高温度下转变为全流体状的l(d)相。随着压力增加,所有相界向上移动到更高温度。发现在所覆盖的温度范围内,筏状l(d)+l(o)两相共存区域的宽度约为2千巴。在更高压力下,检测到向l(o)+s(o)两相区域的转变,并且在更高压力和低温下,检测到全有序的s(o)相。最后,讨论了这些研究在生物学(高压诱导麦角固醇合成上调)和生物技术(真菌微生物失活)方面的相关性。这些结果对于理解膜蛋白在外部应激条件(如高压)下的功能可能特别有趣。有趣的是,在约2千巴的这个压力范围内,在天然膜环境中已观察到膜蛋白功能的停止。当质膜变得过于刚性,即达到整体有序的构象状态时,它们可能在压力/温度条件下失活。