Liu S S, Wu J H, Li Q H
State Key Laboratory of Membrane & Membrane Biotechnology, Institute of Zoology, Academia Sinica, Beijing.
Shi Yan Sheng Wu Xue Bao. 1995 Jun;28(2):137-45.
By using resonance energy transfer assay (RET) and fluorescence microscopy we show experimental evidence that membrane fusion of Ehrlich ascites of mastocarcinoma cells with liposomes could be induced by the proton translocation activity associated with NADH-ferricyanide redox enzyme of transplasma membrane of cancer cells. The iodoacetate, an inhibitor of glycolysis, was found to be able to depress the proton translocation activity and also to inhibit the membrane fusion. It is suggested that NADH produced mainly by glycolysis is utilized as the substrate (electron donor) for transmembrane ferricyanide reduction, and the proton pumping activity in the cancer cells is coupled to the transmembrane NADH-ferricyanide redox enzyme system. Experiments also show that membrane fusion extent of cancer cells with liposomes is proportional to the amount of H+ pumped out by the cells and membrane fusion process also exhibits a H+ consuming mode just as in fusion process of mitochondria with liposomes by redox enzyme proton pumps of respiratory chain. All of the results presented in this paper consists with recent reports of this laboratory, which indicated that various types of proton pumping system from different membrane system of cell have a new function in membrane fusion. Therefore, the proton pumping induced membrane fusion may have a more general physiological importance in triggering and modulating fusion process of native membrane in vivo.
通过使用共振能量转移分析(RET)和荧光显微镜,我们展示了实验证据,即艾氏腹水乳腺癌细胞与脂质体的膜融合可由与癌细胞跨质膜的NADH-铁氰化物氧化还原酶相关的质子转运活性诱导。发现糖酵解抑制剂碘乙酸能够抑制质子转运活性并抑制膜融合。这表明主要由糖酵解产生的NADH被用作跨膜铁氰化物还原的底物(电子供体),并且癌细胞中的质子泵活性与跨膜NADH-铁氰化物氧化还原酶系统偶联。实验还表明,癌细胞与脂质体的膜融合程度与细胞泵出的H +量成正比,并且膜融合过程也表现出H +消耗模式,就像线粒体通过呼吸链的氧化还原酶质子泵与脂质体融合过程一样。本文提出的所有结果与该实验室最近的报告一致,该报告表明来自细胞不同膜系统的各种类型的质子泵系统在膜融合中具有新功能。因此,质子泵诱导的膜融合在触发和调节体内天然膜的融合过程中可能具有更普遍的生理重要性。