Sherwood D, Montal M
Department ofBiochemistry, Centro de Investigación y de Estudios Avanzadosdel Instituto Politécnico Nacional, Mexico City, Mexico.
Biophys J. 1975 May;15(5):417-34. doi: 10.1016/S0006-3495(75)85827-9. Epub 2009 Jan 1.
Planar asymmetric bilayer membranes, formed by apposing a monolayer of the neutral lipid glyceroldioleate (GDO) with one of the negatively charged lipid oleyl acid phosphate (OAP), were used to measure the rate of transmembrane OAP migration. The assay for this lipid flip-flop was the interaction of Ca(2+) ions with negatively charged lipids which causes membranes to break: when Ca(2+) is added to the compartment limited initially by the neutral lipid, flip-flop of the charged lipid eventually results in membrane breakdown. At 22+/-2 degrees C, in the absence of an externally applied electric field, an upper limit to the half time of OAP flip-flop was measured as 18.7 h, with a tentative lower limit of 14.4 h.
通过将中性脂质甘油二油酸酯(GDO)的单层与带负电荷的脂质油酰酸磷酸酯(OAP)之一并置形成的平面不对称双层膜,用于测量跨膜OAP迁移的速率。这种脂质翻转的测定方法是Ca(2+)离子与带负电荷的脂质的相互作用,这会导致膜破裂:当将Ca(2+)添加到最初由中性脂质限制的隔室中时,带电脂质的翻转最终会导致膜破裂。在22±2摄氏度下,在没有外部施加电场的情况下,OAP翻转半衰期的上限测量为18.7小时,暂定下限为14.4小时。