Bar L K, Barenholz Y, Thompson T E
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908.
Biochemistry. 1987 Aug 25;26(17):5460-5. doi: 10.1021/bi00391a037.
Spontaneous cholesterol exchange between small unilamellar vesicles comprised of different phospholipids and their binary mixtures has been studied in order to understand the factors involved in the establishment and maintenance of intracellular cholesterol distributions. Exchange was performed from neutral donor vesicles containing different cholesterol concentrations, traces of [3H]cholesterol, and [14C]cholesteryl oleate as a nonexchangeable marker. The acceptor vesicles, in 10-fold excess, had the same composition, but 15 mol % phosphatidylglycerol was included to permit chromatographic separation. Data were best fitted by a single exponential and a base value. In donor vesicles containing only one phospholipid, the kinetic rate constants agreed with data reported previously; however, the base values were larger than the expected equilibrium value of 9.09%. The size of this nonexchangeable pool and the exchange rate were found to depend on the type of phospholipid. In binary phospholipid donor systems, well above the transition temperatures of the lipid components, the exchange parameters were preferentially closer to those of one component according to the order POPC greater than DMPC greater than DPPC greater than bovine brain SPM.
为了了解参与细胞内胆固醇分布建立和维持的因素,对由不同磷脂及其二元混合物组成的小单层囊泡之间的自发胆固醇交换进行了研究。交换是从中性供体囊泡进行的,这些囊泡含有不同的胆固醇浓度、微量的[3H]胆固醇以及作为不可交换标记物的[14C]胆固醇油酸酯。受体囊泡过量10倍,具有相同的组成,但包含15摩尔%的磷脂酰甘油以允许色谱分离。数据最适合用单指数和一个基值来拟合。在仅含有一种磷脂的供体囊泡中,动力学速率常数与先前报道的数据一致;然而,基值大于预期的平衡值9.09%。发现这个不可交换池的大小和交换速率取决于磷脂的类型。在二元磷脂供体系统中,远高于脂质成分的转变温度时,根据POPC大于DMPC大于DPPC大于牛脑SPM的顺序,交换参数优先更接近其中一种成分的参数。