Quantitative Study of the Interactome of PKC味 Involved in the EGF-induced Tumor Cell Chemotaxis
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- 作者：Ruibing Chen ; Yanping Wang ; Yan Liu ; Qing Zhang ; Xiaofang Zhang ; Fei Zhang ; Chia-Hui Paul Shieh ; De Yang ; Ning Zhang
- 刊名：Journal of Proteome Research
- 出版年：2013
- 出版时间：March 1, 2013
- 年：2013
- 卷：12
- 期：3
- 页码：1478-1486
- 全文大小：410K
- 年卷期：v.12,no.3(March 1, 2013)
- ISSN：1535-3907
文摘
Chemotaxis plays an important role in metastasis. In our previous studies, we reported that protein kinase C 味 (PKC味) mediated cancer cell chemotaxis by regulating cytoskeleton rearrangement and cell adhesion. To further study the molecular mechanism of chemotaxis, mass spectrometry-based approaches were employed to investigate the interactome of PKC味 and its changes upon stimulation by epidermal growth factor (EGF). As a result, 233 proteins were identified as potential PKC味 binding partners. Label free quantification was applied to examine the quantitative changes of these interactions involved in the EGF induced chemotaxis. Fifteen identified proteins were enriched and 9 proteins were reduced in the presence of EGF (鈮?.5 folds, p 鈮?0.05). The interaction between cofilin-1 (CFL1) and PKC味 was evidenced and this interaction was enhanced in the EGF induced chemotaxis signaling transduction. In addition, novel PKC味 interacting proteins potentially related with chemotaxis were characterized, such as isoform 1 of nucleophosmin (NPM1). Furthermore, Western blotting and chemotaxis assays were also applied to validate the proteomics result and explore its biological implications. Collectively, the combination of quantitative proteomics and biological assays provides a powerful strategy for elucidating the signaling pathway of tumor cell chemotaxis.