Rapid phosphorylation and reorganization of ezrin and spectrin accompany morphological changes induced in A-431 cells by epidermal growth factor.
Rapid phosphorylation and reorganization of ezrin and spectrin accompany morphological changes induced in A-431 cells by epidermal growth factor.
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DOI：
10.1083/jcb.108.3.921
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发表时间：
1989-03
期刊：
影响因子：
--
通讯作者：
Bretscher A
中科院分区：
文献类型：
--
作者：
Bretscher A
已知向人类癌A-431细胞中添加EGF会在大约2分钟后诱导细胞膜波纹（Chinkers，M.，J.A.McKanna，和S.Cohen.1979.J.CellBiol.83：260-265）和称为p81的蛋白质的磷酸化，p81是多种蛋白质酪氨酸激酶的已知底物（Cooper，J.A.，D.F. Bowen-Pope、E. Raines、R. Ross 和 T. Hunter 1982。《细胞》31：263-273）。 Ezrin是分离的鸡微绒核的大约80,000个细胞骨架蛋白，存在于多种细胞的含肌动蛋白的细胞表面结构中(Bretscher，A.1983.J.Cell Biol.97：425-432)。然后发现 Ezrin 与 p81 同源，并且响应 EGF 而在酪氨酸上磷酸化（Gould，K.L.，J.A.Cooper，A.Bretscher，和 T.Hunter.1986.J.Cell Biol.102：660-669）。在此，描述了从人胎盘中纯化埃兹蛋白。使用人埃兹蛋白抗体以及其他微丝相关蛋白的抗体来追踪 EGF 处理后这些蛋白在 A-431 细胞中的分布和磷酸化。 EGF 在 30 秒内诱导这些细胞上微绒毛样表面结构的形成，并且在添加 EGF 后大约 2-5 分钟，这些结构就会变成膜褶皱；大约 10-20 分钟后，细胞就会聚集起来。埃兹蛋白被募集到微绒毛样结构和膜褶皱中，并在与这些表面结构的形成和消失平行的时间过程中在酪氨酸和丝氨酸上被磷酸化。血影蛋白被募集到膜皱褶中，并显示出类似的快速磷酸化动力学，但仅针对丝氨酸残基，并且在细胞的聚集过程中保持磷酸化。微绒毛状结构和膜褶边也富含纤维蛋白和α-肌动蛋白。肌球蛋白迅速重组成条纹图案，这与其在细胞变圆中发挥的作用一致。这些结果表明，两种皮质蛋白，埃兹蛋白和血影蛋白，在与细胞表面重塑一致的时间过程中被磷酸化。结果与埃兹蛋白磷酸化可能在细胞表面突起的形成中发挥作用而血影蛋白磷酸化可能参与细胞表面更平坦区域的重塑的观点一致。
Addition of EGF to human carcinoma A-431 cells is known to induce membrane ruffling after approximately 2 min (Chinkers, M., J. A. McKanna, and S. Cohen. 1979. J. Cell Biol. 83:260-265) and the phosphorylation of a protein referred to as p81, a known substrate for various protein-tyrosine kinases (Cooper, J. A., D. F. Bowen-Pope, E. Raines, R. Ross, and T. Hunter. 1982. Cell. 31:263-273). Ezrin, a Mr approximately 80,000 cytoskeletal protein of the isolated chicken microvillar core, is present in actin-containing cell surface structures of a wide variety of cells (Bretscher, A. 1983. J. Cell Biol. 97:425-432). Ezrin was then found to be homologous to p81 and to be phosphorylated on tyrosine in response to EGF (Gould, K. L., J. A. Cooper, A. Bretscher, and T. Hunter. 1986. J. Cell Biol. 102:660-669). Here, the purification of ezrin from human placenta is described. Antibodies to human ezrin, together with antibodies to other microfilament-associated proteins, were used to follow the distribution and phosphorylation of these proteins in A-431 cells after EGF treatment. EGF induces the formation of microvillar-like surface structures on these cells within 30 s and these give way to membrane ruffles at approximately 2-5 min after EGF addition; the cells then round up after approximately 10-20 min. Ezrin is recruited into the microvillar-like structures and the membrane ruffles, and is phosphorylated on tyrosine and serine in a time course that parallels the formation and disappearance of these surface structures. Spectrin is recruited into the membrane ruffles and shows a similar rapid kinetics of phosphorylation, but only on serine residues, and remains phosphorylated through the rounding up of the cells. The microvillar- like structures and membrane ruffles are also enriched in fimbrin and alpha-actinin. Myosin becomes rapidly reorganized into a striated pattern that is consistent with it playing a role in cell rounding. These results show that two cortical proteins, ezrin and spectrin, become phosphorylated in a time course coincident with remodeling of the cell surface. The results are consistent with the notion that ezrin phosphorylation may play a role in the formation of cell surface projections whereas spectrin phosphorylation may be involved in remodelling of more planar areas of the cell surface.
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影响因子：
7.8
作者：
BRETSCHER, A;LYNCH, W
通讯作者：
LYNCH, W
影响因子：
64.5
作者：
HUANG, KS;WALLNER, BP;PEPINSKY, RB
通讯作者：
PEPINSKY, RB
DOI：
10.1083/jcb.83.1.82
发表时间：
1979-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Haigler HT;McKanna JA;Cohen S
通讯作者：
Cohen S
DOI：
10.1083/jcb.86.1.335
发表时间：
1980-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bretscher A;Weber K
通讯作者：
Weber K
DOI：
10.1073/pnas.74.2.565
发表时间：
1977-01-01
影响因子：
11.1
作者：
FABRICANT, RN;DELARCO, JE;TODARO, GJ
通讯作者：
TODARO, GJ