Schneider E, Haest C W, Plasa G, Deuticke B
Biochim Biophys Acta. 1986 Mar 13;855(3):325-36. doi: 10.1016/0005-2736(86)90078-7.
Incorporation of the channel-forming polyene antibiotic amphotericin B and of cytotoxins from Staphylococcus aureus (alpha-toxin) or Pseudomonas aeruginosa into erythrocyte membranes results in a concentration-dependent enhancement of the flip rates of exogenous lysophosphatidylcholine. The flip rate is also enhanced by incorporation of tetracaine and dibucaine. Removal of tetracaine and amphotericin B from the cells normalizes the flip rates. In parallel to the enhancement of flip rates, alpha-toxin produces a loss of transmembrane asymmetry of both phosphatidylethanolamine and phosphatidylserine. Pretreatment of cells with amphotericin or high concentrations (over 2.5 mmol . l-1) of tetracaine, followed by removal of the perturbing agent by washing, produces a selective loss of the asymmetric orientation of phosphatidylethanolamine to the inner membrane layer, as evaluated by the accessibility of the lipid towards cleavage by phospholipase A2. The extent to which asymmetry is lost depends on the time of pretreatment with amphotericin or tetracaine, indicating a limitation by the rate of reorientation of phosphatidylethanolamine to the outer membrane surface. Evaluation of the accessibility of phosphatidylethanolamine towards cleavage by phospholipase A2 in the presence of local anesthetics indicates accessible fractions much higher than those obtained after removal of the perturbant. In the presence of tetracaine, endofacial phosphatidylethanolamine seems somehow to become accessible to phospholipase A2. Phosphatidylserine does not exhibit this peculiarity. The results indicate that various types of perturbation of the lipid domain of the erythrocyte membrane may enhance the transbilayer mobility of phospholipids as well as destabilize the asymmetric distribution of aminophospholipids. However, as in other instances reported previously (Haest, C.W.M., Erusalimsky, J., Dressler, V., Kunze, I. and Deuticke B. (1983) Biomed. Biochim. Acta 42, 17-21), there is no tight coupling between transbilayer mobility and destabilization of asymmetry of the transbilayer distribution of phospholipids.
将形成通道的多烯抗生素两性霉素B以及来自金黄色葡萄球菌（α-毒素）或铜绿假单胞菌的细胞毒素掺入红细胞膜中，会导致外源性溶血磷脂酰胆碱翻转速率呈浓度依赖性增加。丁卡因和布比卡因的掺入也会提高翻转速率。从细胞中去除丁卡因和两性霉素B可使翻转速率恢复正常。与翻转速率增加同时，α-毒素会导致磷脂酰乙醇胺和磷脂酰丝氨酸的跨膜不对称性丧失。用两性霉素或高浓度（超过2.5 mmol·l-1）的丁卡因预处理细胞，然后通过洗涤去除干扰剂，通过脂质对磷脂酶A2裂解的可及性评估，会导致磷脂酰乙醇胺向内膜层的不对称取向选择性丧失。不对称性丧失的程度取决于用两性霉素或丁卡因预处理的时间，这表明磷脂酰乙醇胺重新定向到外膜表面的速率存在限制。在局部麻醉剂存在下评估磷脂酰乙醇胺对磷脂酶A2裂解的可及性，表明可及部分远高于去除干扰剂后获得的部分。在丁卡因存在下，内膜面的磷脂酰乙醇胺似乎以某种方式变得可被磷脂酶A2作用。磷脂酰丝氨酸不表现出这种特性。结果表明，红细胞膜脂质结构域的各种类型的扰动可能会增强磷脂的跨双层流动性，并破坏氨基磷脂的不对称分布。然而，正如之前报道的其他情况（海斯特，C.W.M.，叶鲁萨利姆斯基，J.，德雷斯勒，V.，昆泽，I.和多伊蒂克，B.（1983年）《生物医学与生物化学学报》42，17 - 21）一样，跨双层流动性与磷脂跨双层分布不对称性的破坏之间没有紧密的耦合关系。
