Lipid recycling between the plasma membrane and intracellular compartments: transport and metabolism of fluorescent sphingomyelin analogues in cultured fibroblasts.
Lipid recycling between the plasma membrane and intracellular compartments: transport and metabolism of fluorescent sphingomyelin analogues in cultured fibroblasts.
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DOI：
10.1083/jcb.108.6.2169
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发表时间：
1989-06
期刊：
影响因子：
--
通讯作者：
Pagano RE
中科院分区：
文献类型：
--
作者：
Koval M;Pagano RE
在中国仓鼠卵巢(CHO-K1)成纤维细胞单层中，我们研究了神经鞘磷脂的荧光类似物D-红血球和L-苏体在中国仓鼠卵巢成纤维细胞中的代谢和细胞内转运。C6-NBD-SM通过将C6-NBD-SM单体从脂质体转移到细胞中，在7℃下整合到质膜双层中。细胞洗涤，在37℃升温10-15分钟内，C6-NBD-SM从质膜内化到与中心粒共定位的核周位置，与溶酶体和高尔基体不同。这个核周区域也被内化的罗丹明结合的转铁蛋白标记。当微管被诺康唑破坏时，C6-NBD-SM的内吞作用并没有被抑制；相反，荧光脂质分布在细胞周围的小泡中，而不是内化到细胞的核周区域。还考察了C6-NBD-SM在37℃时对其他荧光鞘脂的代谢及其对C6-NBD-SM转运的影响。为了研究质膜脂循环，首先将C6-NBD-SM插入CHO-K1细胞的质膜，然后允许细胞在37℃内化，然后在7℃与非荧光脂质体孵育去除质膜上的C6-NBD-SM，使细胞只含有内化的荧光脂质。然后观察到C6-NBD-SM在进一步37℃孵育后从细胞内隔间返回到质膜。完整的C6-NBD-SM在质膜和胞内室之间循环的半衰期约为40分钟。莫能菌素或诺可达唑对细胞C6-NBD-SM循环无抑制作用。
We examined the metabolism and intracellular transport of the D-erythro and L-threo stereoisomers of a fluorescent analogue of sphingomyelin, N- (N-[6-[(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] caproyl])- sphingosylphosphorylcholine (C6-NBD-SM), in Chinese hamster ovary (CHO- K1) fibroblast monolayers. C6-NBD-SM was integrated into the plasma membrane bilayer by transfer of C6-NBD-SM monomers from liposomes to cells at 7 degrees C. The cells were washed, and within 10-15 min of being warmed to 37 degrees C, C6-NBD-SM was internalized from the plasma membrane to a perinuclear location that colocalized with the centriole and was distinct from the lysosomes and the Golgi apparatus. This perinuclear region was also labeled by internalized rhodamine- conjugated transferrin. C6-NBD-SM endocytosis was not inhibited when the microtubules were disrupted with nocodazole; rather, the fluorescent lipid was distributed in vesicles throughout the cell periphery instead of being internalized to the perinuclear region of the cell. The metabolism of C6-NBD-SM to other fluorescent sphingolipids at 37 degrees C and its effect on C6-NBD-SM transport was also examined. To study plasma membrane lipid recycling, C6-NBD-SM was first inserted into the plasma membrane of CHO-K1 cells and then allowed to be internalized by the cells at 37 degrees C. Any C6-NBD-SM remaining at the plasma membrane was then removed by incubation with nonfluorescent liposomes at 7 degrees C, leaving cells containing only internalized fluorescent lipid. The return of C6-NBD-SM to the plasma membrane from intracellular compartments upon further 37 degrees C incubation was then observed. The half-time for a complete round C6-NBD- SM recycling between the plasma membrane and intracellular compartments was approximately 40 min. Pretreatment of cells with either monensin or nocodazole did not inhibit C6-NBD-SM recycling.
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溶酶体膜的糖蛋白。
影响因子：
7.8
作者：
LEWIS, V;GREEN, SA;MARSH, M;VIHKO, P;HELENIUS, A;MELLMAN, I
通讯作者：
MELLMAN, I
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2.9
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影响因子：
13.8
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HOPKINS, CR
影响因子：
3.4
作者：
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通讯作者：
KISHIMOTO, Y
影响因子：
7.8
作者：
FERRIS, AL;BROWN, JC;STORRIE, B
通讯作者：
STORRIE, B