Identification of dynamic changes in proteins associated with the cellular cytoskeleton after exposure to okadaic acid.
Identification of dynamic changes in proteins associated with the cellular cytoskeleton after exposure to okadaic acid.
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DOI：
10.3390/md11061763
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发表时间：
2013-05-24
期刊：
影响因子：
5.4
通讯作者：
Fladmark KE
中科院分区：
文献类型：
--
作者：
Opsahl JA;Ljostveit S;Solstad T;Risa K;Roepstorff P;Fladmark KE
细胞暴露于贝类毒素冈田酸，导致细胞骨架结构的急剧重组和细胞-细胞接触的丧失。当细胞暴露于高浓度的冈田酸（100-500 nM）时，形态学重排之后是凋亡性细胞死亡。冈田酸抑制广泛作用的Ser/Thr蛋白磷酸酶1和2A，这导致大量蛋白质的过度磷酸化。这些过度磷酸化的蛋白质中的一些最有可能是冈田酸诱导的细胞形态重组中的关键参与者。我们想鉴定这些磷蛋白，并在细胞脂筏中寻找它们，这些脂筏已被发现含有调节细胞骨架动力学和细胞粘附的蛋白质。通过在细胞培养物中使用氨基酸的稳定同位素标记，用冈田酸（400 nM）处理的细胞可以在分离脂筏之前与对照细胞组合。通过质谱鉴定冈田酸诱导的蛋白磷酸化事件和易位。冈田酸被证明可以调节与肌动蛋白细胞骨架、微管和细胞粘附结构相关的蛋白质的磷酸化状态和位置。大量的这些冈田酸调节的蛋白质以前也被证明是类似的细胞增殖和迁移前的调节。我们的研究结果表明，冈田酸激活一般的细胞信号转导途径，诱导皮质肌动蛋白细胞骨架和细胞脱离的崩溃。
Exposure of cells to the diarrhetic shellfish poison, okadaic acid, leads to a dramatic reorganization of cytoskeletal architecture and loss of cell-cell contact. When cells are exposed to high concentrations of okadaic acid (100–500 nM), the morphological rearrangement is followed by apoptotic cell death. Okadaic acid inhibits the broad acting Ser/Thr protein phosphatases 1 and 2A, which results in hyperphosphorylation of a large number of proteins. Some of these hyperphosphorylated proteins are most likely key players in the reorganization of the cell morphology induced by okadaic acid. We wanted to identify these phosphoproteins and searched for them in the cellular lipid rafts, which have been found to contain proteins that regulate cytoskeletal dynamics and cell adhesion. By using stable isotope labeling by amino acids in cell culture cells treated with okadaic acid (400 nM) could be combined with control cells before the isolation of lipid rafts. Protein phosphorylation events and translocations induced by okadaic acid were identified by mass spectrometry. Okadaic acid was shown to regulate the phosphorylation status and location of proteins associated with the actin cytoskeleton, microtubules and cell adhesion structures. A large number of these okadaic acid-regulated proteins have previously also been shown to be similarly regulated prior to cell proliferation and migration. Our results suggest that okadaic acid activates general cell signaling pathways that induce breakdown of the cortical actin cytoskeleton and cell detachment.
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DOI：
10.2741/dawson
发表时间：
1999-10-01
期刊：
Frontiers in Bioscience
影响因子：
--
作者：
Dawson, John F.;Holmes, Charles F. B.
通讯作者：
Holmes, Charles F. B.
影响因子：
2.4
作者：
Hanana H;Talarmin H;Pennec JP;Droguet M;Morel J;Dorange G
通讯作者：
Dorange G
影响因子：
3.3
作者：
Bordeleau F;Galarneau L;Gilbert S;Loranger A;Marceau N
通讯作者：
Marceau N
影响因子：
6
作者：
Ito, A;Koma, YI;Kitamura, Y
通讯作者：
Kitamura, Y
影响因子：
3.7
作者：
Finlayson AE;Freeman KW
通讯作者：
Freeman KW