Lipid domain structure of the plasma membrane revealed by patching of membrane components.
Lipid domain structure of the plasma membrane revealed by patching of membrane components.
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DOI：
10.1083/jcb.141.4.929
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发表时间：
1998-05-18
影响因子：
7.8
通讯作者：
Simons, K
中科院分区：
文献类型：
--
作者：
Harder, T;Scheiffele, P;Verkade, P;Simons, K
糖脂和胆固醇的侧向组装，即“筏”，已被认为在细胞过程中发挥作用，如膜分离、信号转导和细胞黏附。我们研究了非极化细胞质膜中RAFT结构域的结构。高表达的细胞膜标志物均匀分布于细胞膜。我们比较了RAFT标记对(由Triton X-100中的不溶性定义)和RAFT/非RAFT标记对的修补行为。为此，我们使用抗体和/或霍乱毒素将糖基化磷脂酰肌醇(GPI)锚定蛋白胎盘碱性磷酸酶(PLAP)、Thy-1、流感病毒血凝素(HA)和RAFT脂质神经节苷脂GM1连接起来。这些RAFT标记的补丁在BHK细胞和Jurkat T淋巴瘤细胞中广泛重叠。重要的是，GPI锚定的PLAP片断积累了src样蛋白酪氨酸激酶fyn，被认为锚定在RAFT结构域的细胞质小叶中。相反，作为非RAFT标记物的补丁RAFT组分和转铁蛋白受体斑块被明显分离。综上所述，我们的数据有力地表明，交联型RAFT元件的结合是由它们共同的脂类环境介导的，而RAFT和非RAFT斑块的分离是由不同脂相的不相容引起的。这一观点得到了这样一个发现的支持，即胆固醇消耗消除了隔离。我们的结果与这样的观点是一致的，即非极化细胞质膜上的RAFT结构域通常很小且高度分散，但RAFT的大小可以通过RAFT组分的齐聚来调节。
Lateral assemblies of glycolipids and cholesterol, “rafts,” have been implicated to play a role in cellular processes like membrane sorting, signal transduction, and cell adhesion. We studied the structure of raft domains in the plasma membrane of non-polarized cells. Overexpressed plasma membrane markers were evenly distributed in the plasma membrane. We compared the patching behavior of pairs of raft markers (defined by insolubility in Triton X-100) with pairs of raft/non-raft markers. For this purpose we cross-linked glycosyl-phosphatidylinositol (GPI)-anchored proteins placental alkaline phosphatase (PLAP), Thy-1, influenza virus hemagglutinin (HA), and the raft lipid ganglioside GM1 using antibodies and/or cholera toxin. The patches of these raft markers overlapped extensively in BHK cells as well as in Jurkat T–lymphoma cells. Importantly, patches of GPI-anchored PLAP accumulated src-like protein tyrosine kinase fyn, which is thought to be anchored in the cytoplasmic leaflet of raft domains. In contrast patched raft components and patches of transferrin receptor as a non-raft marker were sharply separated. Taken together, our data strongly suggest that coalescence of cross-linked raft elements is mediated by their common lipid environments, whereas separation of raft and non-raft patches is caused by the immiscibility of different lipid phases. This view is supported by the finding that cholesterol depletion abrogated segregation. Our results are consistent with the view that raft domains in the plasma membrane of non-polarized cells are normally small and highly dispersed but that raft size can be modulated by oligomerization of raft components.
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影响因子：
7.5
作者：
Harder, T;Simons, K
通讯作者：
Simons, K
影响因子：
4.8
作者：
HANADA, K;NISHIJIMA, M;PAGANO, RE
通讯作者：
PAGANO, RE
DOI：
10.1073/pnas.92.19.8655
发表时间：
1995-09-12
影响因子：
11.1
作者：
FRA, AM;WILLIAMSON, E;PARTON, RG
通讯作者：
PARTON, RG
DOI：
10.1083/jcb.140.6.1357
发表时间：
1998-03-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Keller P;Simons K
通讯作者：
Simons K
影响因子：
2.9
作者：
HARRIS, JS;EPPS, DE;KEZDY, FJ
通讯作者：
KEZDY, FJ