Andreoli T E, Bangham J A, Tosteson D C
J Gen Physiol. 1967 Jul;50(6):1729-49. doi: 10.1085/jgp.50.6.1729.
Lipids were obtained from high potassium (HK) and low potassium (LK) sheep red cells by sequential extraction of the erythrocytes with isopropanol-chloroform, chloroform-methanol-0.1 M KCl, and chloroform. The extract contained cholesterol and phospholipid in a molar ratio of 0.8:1.0, and less than 1% protein contaminant. Stable thin lipid membranes separating two aqueous compartments were formed from an erythrocyte lipid-hydrocarbon solution, and had an electrical resistance of approximately 10(8) ohm-cm(2) and a capacitance of 0.38-0.4 microf/cm(2). From the capacitance values, membrane thickness was estimated to be 46-132 A, depending on the assumed value for the dielectric constant (2.0-4.5). Membrane voltage was recorded in the presence of ionic (NaCl and/or KCl) concentration gradients in the solutions bathing the membrane. The permeability of the membrane to Na(+), K(+), and Cl(-) (expressed as the transference number, T(ion)) was computed from the steady-state membrane voltage and the activity ratio of the ions in the compartments bathing the membrane. T(Na) and T(K) were approximately equal ( approximately 0.8) and considerably greater than T(Cl) ( approximately 0.2). The ionic transference numbers were independent of temperature, the hydrocarbon solvent, the osmolarity of the solutions bathing the membranes, and the cholesterol content of the membranes, over the range 21-38 degrees C. The high degree of membrane cation selectivity was tentatively attributed to the negatively charged phospholipids (phosphatidylethanolamine and phosphatidylserine) present in the lipid extract.
通过用异丙醇 - 氯仿、氯仿 - 甲醇 - 0.1M氯化钾和氯仿依次提取红细胞,从高钾(HK)和低钾(LK)绵羊红细胞中获得脂质。提取物中胆固醇与磷脂的摩尔比为0.8:1.0,蛋白质污染物含量低于1%。由红细胞脂质 - 烃溶液形成了分隔两个水相区室的稳定薄脂质膜,其电阻约为10⁸欧姆·厘米²,电容为0.38 - 0.4微法/厘米²。根据电容值,膜厚度估计为46 - 132埃,具体取决于介电常数的假定值(2.0 - 4.5)。在膜两侧溶液存在离子(氯化钠和/或氯化钾)浓度梯度的情况下记录膜电压。根据稳态膜电压和膜两侧溶液中离子的活度比计算膜对钠离子、钾离子和氯离子的渗透率(以迁移数T(离子)表示)。T(Na)和T(K)大致相等(约为0.8),且远大于T(Cl)(约为0.2)。在21 - 38摄氏度范围内,离子迁移数与温度、烃类溶剂、膜两侧溶液的渗透压以及膜的胆固醇含量无关。膜的高度阳离子选择性初步归因于脂质提取物中存在的带负电荷的磷脂(磷脂酰乙醇胺和磷脂酰丝氨酸)。