Widomska Justyna, Raguz Marija, Dillon James, Gaillard Elizabeth R, Subczynski Witold K
Department of Biophysics, Medical College of Wisconsin, Milwaukee, 8701 Watertown Plank Road, WI 53226, USA.
Biochim Biophys Acta. 2007 Jun;1768(6):1454-65. doi: 10.1016/j.bbamem.2007.03.007. Epub 2007 Mar 20.
The physical properties of a membrane derived from the total lipids of a calf lens were investigated using EPR spin labeling and were compared with the properties of membranes made of an equimolar 1-palmitoyl-2-oleoylphosphatidylcholine/cholesterol (POPC/Chol) mixture and of pure POPC. Conventional EPR spectra and saturation-recovery curves show that spin labels detect a single homogenous environment in all three membranes. Profiles of the order parameter, hydrophobicity, and oxygen transport parameter are practically identical in lens lipid and POPC/Chol membranes, but differ drastically from profiles in pure POPC membranes. In both lens lipid and POPC/Chol membranes, the lipids are strongly immobilized at all depths, which is in contrast to the high fluidity of the POPC membrane. Hydrophobicity and oxygen transport parameter profiles in lens lipid and POPC/Chol membranes have a rectangular shape with an abrupt change between the C9 and C10 positions, which is approximately where the steroid ring structure of cholesterol reaches into the membrane. At this position, hydrophobicity increases from the level of methanol to the level of hexane, and the oxygen transport parameter increases by a factor of 2-3. These profiles in POPC membranes are bell-shaped. It is concluded that the high level of cholesterol in lens lipids makes the membrane stable, immobile, and impermeable to both polar and nonpolar molecules.
利用电子顺磁共振(EPR)自旋标记法研究了从小牛晶状体总脂质中提取的膜的物理性质,并将其与由等摩尔的1-棕榈酰-2-油酰磷脂酰胆碱/胆固醇(POPC/Chol)混合物制成的膜以及纯POPC膜的性质进行了比较。传统的EPR光谱和饱和恢复曲线表明,自旋标记在所有三种膜中检测到单一的均匀环境。在晶状体脂质膜和POPC/Chol膜中,序参数、疏水性和氧传输参数的分布实际上是相同的,但与纯POPC膜中的分布有很大差异。在晶状体脂质膜和POPC/Chol膜中,脂质在所有深度都被强烈固定,这与POPC膜的高流动性形成对比。晶状体脂质膜和POPC/Chol膜中的疏水性和氧传输参数分布呈矩形,在C9和C10位置之间有突然变化,这大约是胆固醇的类固醇环结构伸入膜内的位置。在这个位置,疏水性从甲醇水平增加到己烷水平,氧传输参数增加2至3倍。POPC膜中的这些分布呈钟形。结论是,晶状体脂质中高水平的胆固醇使膜稳定、不流动,并且对极性和非极性分子均不可渗透。