Profiling the interactome of protein kinase C ζ by proteomics and bioinformatics.
Profiling the interactome of protein kinase C ζ by proteomics and bioinformatics.
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通过蛋白质组学和生物信息学分析蛋白激酶 C zeta 的相互作用组
DOI：
10.1186/s12953-018-0134-8
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发表时间：
2018
期刊：
影响因子：
2
通讯作者：
Chen R
中科院分区：
文献类型：
--
作者：
Hou C;Li Y;Liu H;Dang M;Qin G;Zhang N;Chen R
蛋白激酶C亚型（PKC β）是非典型蛋白激酶C的一种亚型，是癌症的关键调节因子。然而，PKC β调控肿瘤发生和转移的分子和细胞机制仍不完全清楚。本研究通过蛋白质组学和生物信息学分析，建立了与PKC β相关的蛋白质-蛋白质相互作用（PPI）网络，为进一步了解PKC β的多种生物学作用奠定了基础。通过免疫共沉淀法从乳腺癌细胞MDA-MB-231中纯化与PKC β相关的蛋白复合物，并通过LC-MS/MS进行鉴定。使用CRAPome数据库过滤观察到的蛋白质以消除潜在的假阳性。将蛋白质组学鉴定结果与PPI数据库检索相结合，构建相互作用组网络。利用PANTHER数据库和大卫进行基因本体（GO）和途径分析。接下来，通过免疫共沉淀、Western印迹和免疫荧光验证了PKC β和蛋白磷酸酶2催化亚基α（PPP 2CA）之间的相互作用。并利用TCGA数据库和COSMIC数据库分析这两种蛋白在临床标本中的表达情况。建立了以PKC为中心的PPI网络，包含178个节点和1225个连接。网络分析表明，所鉴定的蛋白质与调节癌症相关细胞过程的几个关键信号通路显著相关。通过结合蛋白质组学和生物信息学分析，构建了一个以PKC β为中心的PPI网络，为PKC β在肿瘤调控和细胞生物学其他方面的生物学作用提供了更全面的信息。本文的在线版本（10.1186/s12953-018-0134-8）包含补充材料，可供授权用户使用。
Protein kinase C ζ (PKCζ), an isoform of the atypical protein kinase C, is a pivotal regulator in cancer. However, the molecular and cellular mechanisms whereby PKCζ regulates tumorigenesis and metastasis are still not fully understood. In this study, proteomics and bioinformatics analyses were performed to establish a protein-protein interaction (PPI) network associated with PKCζ, laying a stepping stone to further understand the diverse biological roles of PKCζ. Protein complexes associated with PKCζ were purified by co-immunoprecipitation from breast cancer cell MDA-MB-231 and identified by LC-MS/MS. Two biological replicates and two technical replicates were analyzed. The observed proteins were filtered using the CRAPome database to eliminate the potential false positives. The proteomics identification results were combined with PPI database search to construct the interactome network. Gene ontology (GO) and pathway analysis were performed by PANTHER database and DAVID. Next, the interaction between PKCζ and protein phosphatase 2 catalytic subunit alpha (PPP2CA) was validated by co-immunoprecipitation, Western blotting and immunofluorescence. Furthermore, the TCGA database and the COSMIC database were used to analyze the expressions of these two proteins in clinical samples. The PKCζ centered PPI network containing 178 nodes and 1225 connections was built. Network analysis showed that the identified proteins were significantly associated with several key signaling pathways regulating cancer related cellular processes. Through combining the proteomics and bioinformatics analyses, a PKCζ centered PPI network was constructed, providing a more complete picture regarding the biological roles of PKCζ in both cancer regulation and other aspects of cellular biology. The online version of this article (10.1186/s12953-018-0134-8) contains supplementary material, which is available to authorized users.
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影响因子：
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作者：
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