Mass spectrometric analysis identifies a cortactin-RCC2/TD60 interaction in mitotic cells.
Mass spectrometric analysis identifies a cortactin-RCC2/TD60 interaction in mitotic cells.
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DOI：
10.1016/j.jprot.2012.01.012
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发表时间：
2012-04-03
影响因子：
3.3
通讯作者：
Fox JW
中科院分区：
文献类型：
--
作者：
Grigera PR;Ma L;Borgman CA;Pinto AF;Sherman NE;Parsons JT;Fox JW
Corprin是一种F-肌动蛋白结合蛋白，其作为支架在板状伪足和侵入伪足形成中调节Arp 2/3介导的肌动蛋白聚合，以及在许多不同细胞类型的细胞迁移和内吞中起作用。鉴于受调控的肌动蛋白聚合对许多细胞过程至关重要的事实，我们启动了一项新的corneumn与细胞蛋白质的相互作用的研究，这可能表明迄今为止未描述的corneumn支持的生物活性。在HEK 293细胞中使用改良的稳定同位素标记细胞培养（SILAC）方法和Flag标记的coronin（F-coronin）作为诱饵，我们鉴定了有限的coronin相互作用，包括几种以前未被鉴定为coronin相关蛋白的蛋白质。其中包括丝氨酸/苏氨酸蛋白磷酸酶2A亚基β（PP 2A-β）和RCC 2/TD 60，一种完成有丝分裂和胞质分裂所需的Rac鸟嘌呤核苷酸交换因子（GEF）。在用抗-RCC 2/TD 60抗体免疫沉淀的细胞裂解物中验证corneumn和RCC 2/TD 60之间的相互作用。此外，corneumen是本地化的免疫荧光在赤道平面的分裂HeLa细胞的区域，其中RCC 2/TD 60先前已被本地化，从而提供支持的复合物含有corneumen和RCC 2/TD 60复合物，可能发挥功能作用的细胞进行有丝分裂。
Cortactin is an F-actin binding protein that functions as a scaffold to regulate Arp2/3 mediated actin polymerization in lamellipodia and invadopodia formation as well as functioning in cell migration and endocytosis of many different cell types. In light of the fact that regulated actin polymerization is critical for many cellular processes we launched a search for novel cortactin interactions with cellular proteins that might indicate heretofore undescribed biological activities supported by cortactin. Using a modified stable isotope labeling in cell culture (SILAC) approach in HEK293 cells and Flag-tagged cortactin (F-cortactin) as bait, we identified a limited set of cortactin interactions including several proteins which have not previously been identified as cortactin associated proteins. Among these were serine/threonine-protein phosphatase 2A subunit beta (PP2A-beta) and RCC2/TD60, a Rac guanine nucleotide exchange factor (GEF) required for completion of mitosis and cytokinesis. The interaction between cortactin and RCC2/TD60 was verified in cell lysates immunoprecitated with anti- RCC2/TD60 antibody. Furthermore, cortactin was localized by immunofluorescence in the equatorial plane of dividing HeLa cells in the region where RCC2/TD60 has previously been localized thus providing support for a complex containing cortactin and RCC2/TD60 complex that may play a functional role in cells undergoing mitosis.
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影响因子：
64.8
作者：
Gavin, AC;Bösche, M;Superti-Furga, G
通讯作者：
Superti-Furga, G
DOI：
10.1083/jcb.200405155
发表时间：
2004-11-22
期刊：
The Journal of cell biology
影响因子：
--
作者：
McHugh B;Krause SA;Yu B;Deans AM;Heasman S;McLaughlin P;Heck MM
通讯作者：
Heck MM
影响因子：
4.4
作者：
Goldfinger, Lawrence E.;Ptak, Celeste;Ginsberg, Mark H.
通讯作者：
Ginsberg, Mark H.
影响因子：
6.6
作者：
Banon-Rodriguez, Inmaculada;Monypenny, James;Anton, Ines M.
通讯作者：
Anton, Ines M.
影响因子：
30.8
作者：
Berkel, Simone;Marshall, Christian R.;Rappold, Gudrun A.
通讯作者：
Rappold, Gudrun A.