Paton J C, May B K, Elliott W H
J Bacteriol. 1978 Aug;135(2):393-401. doi: 10.1128/jb.135.2.393-401.1978.
The phospholipid distribution in the membrane of Bacillus amyloliquefaciens was studied by using phospholipase C (B. cereus), phospholipase A2 (Crotalus), and the nonpenetrating chemical probe trinitrobenzenesulfonic acid. After treatment of intact protoplasts of B. amyloliquefaciens with either phospholipase, about 70% of total membrane phospholipid was hydrolyzed; specifically, about 90, 90, and 30% of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin, respectively. Under these conditions, protoplasts remained intact and sealed. However, when protoplasts that were permeabilized by cold-shock treatment were incubated with either of the phospholipases, up to 80% of cardiolipin was hydrolyzed and phosphatidylglycerol and phosphatidylethanolamine were hydrolyzed virtually to completion. In intact cells, 92% of the phosphatidylethanolamine could be labeled with trinitrobenzenesulfonic acid under conditions in which the reagent did not penetrate the membrane to any significant extent. These results indicate that 70% of total phospholipid of this bacillus exists in the outer half of the bilayer. The distribution of phosphatidylethanolamine in this bilayer is highly asymmetric with it being located predominantly in the outer half. The results with phospholipases suggest that the distributions of cardiolipin and phosphatidylglycerol are also asymmetric but independent confirmation of this is required.
利用磷脂酶C（蜡样芽孢杆菌）、磷脂酶A2（响尾蛇）和非穿透性化学探针三硝基苯磺酸，研究了解淀粉芽孢杆菌细胞膜中的磷脂分布。用任何一种磷脂酶处理解淀粉芽孢杆菌的完整原生质体后，约70%的总膜磷脂被水解；具体而言，磷脂酰乙醇胺、磷脂酰甘油和心磷脂分别约有90%、90%和30%被水解。在这些条件下，原生质体保持完整且封闭。然而，当用冷休克处理使其通透化的原生质体与任何一种磷脂酶一起孵育时，高达80%的心磷脂被水解，磷脂酰甘油和磷脂酰乙醇胺几乎完全被水解。在完整细胞中，在试剂基本上不穿透膜的条件下，92%的磷脂酰乙醇胺可用三硝基苯磺酸标记。这些结果表明，该芽孢杆菌总磷脂的70%存在于双层膜的外半部分。磷脂酰乙醇胺在该双层膜中的分布高度不对称，主要位于外半部分。磷脂酶的结果表明心磷脂和磷脂酰甘油的分布也是不对称的，但需要对此进行独立验证。
