Olaisson H, Mårdh S, Arvidson G
J Biol Chem. 1985 Sep 15;260(20):11262-7.
The transverse distribution of the phospholipids in vesicular H+-translocating membranes prepared from pig gastric mucosa was investigated with the aid of phospholipase C, sphingomyelinase, and trinitrobenzenesulfonic acid. The major part (80-90%) of the phosphatidylcholine and the phosphatidylethanolamine, 60% of the phosphatidylserine, and 45% of the sphingomyelin was located on the external, cytoplasmic side of the vesicle membranes. After treatment with phospholipase C the vesicles still behaved as osmometers and appeared as closed vesicles on the electron micrographs. 31P NMR indicated that the phospholipids in untreated vesicles as well as the unhydrolyzed phospholipids in phospholipase C-treated vesicles were arranged in lamellar structures. The 31P NMR spectrum of untreated vesicles to which Pr3+ ions had been added supported the conclusion that the major part of the membrane phospholipids was located on the external surface of the vesicles. A small fraction of the lipids, 3.6 mol %, was found to consist of glycosphingolipids which occurred at a concentration of 52 nmol/mg of protein.
借助磷脂酶C、鞘磷脂酶和三硝基苯磺酸，对从猪胃黏膜制备的囊泡状H⁺转运膜中磷脂的横向分布进行了研究。磷脂酰胆碱和磷脂酰乙醇胺的大部分（80 - 90%）、磷脂酰丝氨酸的60%以及鞘磷脂的45%位于囊泡膜的外侧，即细胞质一侧。用磷脂酶C处理后，囊泡仍表现为渗透压计，在电子显微镜照片上呈现为封闭的囊泡。³¹P核磁共振表明，未处理囊泡中的磷脂以及磷脂酶C处理囊泡中未水解的磷脂呈层状结构排列。添加了Pr³⁺离子的未处理囊泡的³¹P核磁共振谱支持了膜磷脂大部分位于囊泡外表面的结论。发现一小部分脂质（3.6摩尔%）由糖鞘脂组成，其浓度为52纳摩尔/毫克蛋白质。
