Transbilayer movement of a fluorescent phosphatidylethanolamine analogue across the plasma membranes of cultured mammalian cells.
Transbilayer movement of a fluorescent phosphatidylethanolamine analogue across the plasma membranes of cultured mammalian cells.
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发表时间：
1985-01
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通讯作者：
R. Sleight;R. Pagano
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作者：
R. Sleight;R. Pagano
研究了磷脂酰乙醇胺荧光类似物插入培养的中国仓鼠成纤维细胞质膜后的内化情况。当由 50 mol% 1-酰基-2-(N-4-硝基苯并-2-氧杂-1,3-二唑)-氨基己酰磷脂酰乙醇胺 (C6-NBD-PE) 和二油酰磷脂酰胆碱组成的脂质体与单层细胞培养物在 2℃ 下孵育时，荧光脂质会从脂质体自发转移到细胞。只要细胞保持在 2 摄氏度，荧光脂质就会保留在质膜上。然而，如果在除去荧光脂质体后，将培养物加热至 37°C，则 C6-NBD-PE 会被内化，除了质膜外，还会驻留在核膜、线粒体和高尔基体中。通过向培养介质中添加 2-脱氧葡萄糖和叠氮化钠，可以阻断荧光脂质向高尔基体的递送。有证据表明，虽然荧光脂质向高尔基体的递送主要依赖于内吞作用，但向核膜和线粒体的递送是通过脂质穿过质膜的快速跨双层运动以及随后脂质单体的易位而发生的。研究发现，C6-NBD-PE 穿过质膜的快速跨双层运动是一个温度依赖性过程，在 7 摄氏度以下会被阻止。
The internalization of a fluorescent analogue of phosphatidylethanolamine following its insertion into the plasma membrane of cultured Chinese hamster fibroblasts was examined. When liposomes composed of 50 mol % 1-acyl-2-(N-4-nitrobenzo-2-oxa-1,3-diazole)-aminocaproyl phosphatidylethanolamine (C6-NBD-PE) and dioleoylphosphatidylcholine were incubated with monolayer cell cultures at 2 degrees C, a spontaneous transfer of the fluorescent lipid from liposomes to cells occurred. As long as the cells were kept at 2 degrees C, the fluorescent lipid remained at the plasma membrane. However, if, after removing the fluorescent liposomes, the cultures were warmed to 37 degrees C, the C6-NBD-PE was internalized and resided in the nuclear envelope, mitochondria, and Golgi apparatus in addition to the plasma membrane. Delivery of the fluorescent lipid to the Golgi apparatus could be blocked by the addition of 2-deoxyglucose plus sodium azide to the incubation medium. Evidence is presented suggesting that while delivery of the fluorescent lipid to the Golgi apparatus was mainly dependent on endocytosis, delivery to the nuclear envelope and mitochondria occurred by rapid transbilayer movement of the lipid across the plasma membrane followed by translocation of lipid monomers. Rapid transbilayer movement of C6-NBD-PE across the plasma membrane was found to be a temperature-dependent process that was blocked below 7 degrees C.