TRANSPORT OF EXOGENOUS FLUORESCENT PHOSPHATIDYLSERINE ANALOG TO THE GOLGI-APPARATUS IN CULTURED FIBROBLASTS
TRANSPORT OF EXOGENOUS FLUORESCENT PHOSPHATIDYLSERINE ANALOG TO THE GOLGI-APPARATUS IN CULTURED FIBROBLASTS
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DOI：
10.1083/jcb.113.2.235
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发表时间：
1991-04-01
影响因子：
7.8
通讯作者：
ARAKAWA, Y
中科院分区：
文献类型：
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作者：
KOBAYASHI, T;ARAKAWA, Y
我们研究了培养的成纤维细胞中磷脂酰丝氨酸荧光类似物 1-棕榈酰-2-(N-[12[(7-硝基苯甲酰-2-氧杂-1,3-二唑-4-基)氨基]十二酰基])-磷脂酰丝氨酸 ([棕榈酰-C-12-NBD]-PS) 的细胞内转运和代谢。 当单层培养物与含有(棕榈酰-C-12-NBD)-PS的脂质体在37℃下孵育时，荧光PS被转运至高尔基体。 在相同的实验条件下，含有 NBD 的磷脂酰胆碱、磷脂酰乙醇胺 (PE) 或磷脂酸类似物不会在高尔基体中积累。我们认为这种转运不是由于内吞作用，而是 (棕榈酰-C-12-NBD)-PS 在质膜上掺入和跨双层运动的结果，随后脂质通过非囊泡从质膜易位到高尔基体机制。 细胞 ATP 的消耗会抑制荧光 PS 的摄取，并且会被脂质的结构类似物或用戊二醛或 N-乙基马来酰亚胺预处理细胞所阻断。 掺入细胞后，荧光 PS 被代谢为荧光 PE。 在转换过程中，细胞内荧光的分布发生了变化。 除了高尔基体之外，线粒体也被标记。
We have examined intracellular transport and metabolism of the fluorescent analogue of phosphatidylserine, 1-palmitoyl-2-(N-[12[(7-nitrobenz-2-oxa-1,3-diazole-4-yl)amino]dodecanoyl])-phosphatidylserine ([palmitoyl-C-12-NBD]-PS) in cultured fibroblasts. When monolayer cultures were incubated with liposomes containing (palmitoyl-C-12-NBD)-PS at 37-degrees-C, fluorescent PS was transported to the Golgi apparatus. NBD-containing analogues of phosphatidylcholine, phosphatidylethanolamine (PE), or phosphatidic acid did not accumulate in the Golgi apparatus under the same experimental conditions.We suggest that the transport is not due to endocytosis, but is the result of incorporation and transbilayer movement of the (palmitoyl-C-12-NBD)-PS at the plasma membrane followed by translocation of the lipid from plasma membrane to the Golgi apparatus via nonvesicular mechanisms. Uptake of fluorescent PS was inhibited by depletion of cellular ATP and was blocked by structural analogues of the lipid or by pretreatment of cells with glutaraldehyde or N-ethyl-maleimide. After incorporation into the cell, fluorescent PS was metabolized to fluorescent PE. The intracellular distribution of fluorescence changed during the conversion. In addition to the Golgi apparatus, mitochondria also became labeled.