Rawyler A, van der Schaft P H, Roelofsen B, Op den Kamp J A
Biochemistry. 1985 Mar 26;24(7):1777-83. doi: 10.1021/bi00328a031.
The distribution of phospholipids over outer and inner layers of the plasma membranes of Friend erythroleukemic cells (Friend cells) and mature mouse erythrocytes has been determined. The various techniques which have been applied to establish the phospholipid localization include the following: phospholipase A2, phospholipase C, and sphingomyelinase C treatment, fluorescamine labeling of phosphatidylethanolamine, and a phosphatidylcholine transfer protein mediated exchange procedure. The data obtained with these different techniques were found to be in good agreement with each other. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol were found to be distributed symmetrically over both layers of the plasma membrane of Friend cells. In contrast, sphingomyelin was found to be enriched in the outer layer of the membrane (80-85%), and phosphatidylserine appeared to be present mainly in the inner layer (80-90%). From these results, it was calculated that the outer and inner layers accounted for 46% and 54%, respectively, of the total phospholipid complement of that membrane. Analogous studies on the plasma membrane of mature mouse erythrocytes showed that the transbilayer distribution of the total phospholipid mass appeared to be the same as in the plasma membrane of the Friend cell, namely, 46% and 54% in outer and inner layers, respectively. The outer layer of this membrane contains 57% of the phosphatidylcholine, 20% of the phosphatidylethanolamine, 85% of the sphingomyelin, and 42% of the phosphatidylinositol, and none of the phosphatidylserine was present.(ABSTRACT TRUNCATED AT 250 WORDS)
已确定了弗氏红白血病细胞(弗氏细胞)和成熟小鼠红细胞质膜外层和内层上磷脂的分布情况。用于确定磷脂定位的各种技术如下:磷脂酶A2、磷脂酶C和鞘磷脂酶C处理、磷脂酰乙醇胺的荧光胺标记以及磷脂酰胆碱转移蛋白介导的交换程序。发现用这些不同技术获得的数据彼此吻合良好。结果发现,磷脂酰胆碱、磷脂酰乙醇胺和磷脂酰肌醇在弗氏细胞质膜的两层中呈对称分布。相比之下,鞘磷脂在膜的外层富集(80 - 85%),而磷脂酰丝氨酸似乎主要存在于内层(80 - 90%)。根据这些结果计算得出,该膜外层和内层分别占总磷脂成分的46%和54%。对成熟小鼠红细胞质膜的类似研究表明,总磷脂质量的跨膜分布似乎与弗氏细胞质膜相同,即外层和内层分别为46%和54%。该膜的外层含有57%的磷脂酰胆碱、20%的磷脂酰乙醇胺、85%的鞘磷脂和42%的磷脂酰肌醇,且不存在磷脂酰丝氨酸。(摘要截选至250词)