Proteomic Analysis of the Human Cyclin-dependent Kinase Family Reveals a Novel CDK5 Complex Involved in Cell Growth and Migration
Proteomic Analysis of the Human Cyclin-dependent Kinase Family Reveals a Novel CDK5 Complex Involved in Cell Growth and Migration
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人类细胞周期蛋白依赖性激酶家族的蛋白质组学分析揭示了参与细胞生长和迁移的新型 CDK5 复合物
DOI：
10.1074/mcp.m113.036699
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发表时间：
2014-11-01
影响因子：
7
通讯作者：
Chen, Junjie
中科院分区：
文献类型：
--
作者：
Xu, Shuangbing;Li, Xu;Chen, Junjie
细胞周期蛋白依赖性激酶（Cyclin-dependent kinases，CDKs）是哺乳动物异源二聚体丝氨酸/苏氨酸激酶家族的催化亚基，在细胞周期进程、转录和神经元功能的调控中发挥重要作用。然而，许多CDK的功能、底物和调节知之甚少。为了系统地研究CDK的这些特征，我们对CDK家族进行了蛋白质组学分析，并使用改良的SAINT（Interactome显著性分析）方法在两种不同的细胞系中鉴定了它们相关的蛋白复合物。将质谱数据保存至ProteomeXchange，标识符为PXD 000593和DOI 10.6019/PXD 000593。我们在HEK 293 T细胞中鉴定了753个高置信度候选相互作用蛋白（HCIPs），在MCF 10A细胞中鉴定了352个HCIPs。随后，我们专注于神经元特异性CDK，CDK 5，并发现了两个新的CDK 5结合伙伴，KIAA 0528和成纤维细胞生长因子（酸性）细胞内结合蛋白（FIB），在非神经元细胞。我们发现，这三种蛋白质形成了一个稳定的复合物，KIAA 0528和FIB是组装和稳定复合物所必需的。此外，CDK 5-、KIAA 0528-或FIB-耗尽的乳腺癌细胞显示增殖受损和迁移减少，表明这种复合物是非神经细胞中细胞生长和迁移所需的。我们的研究揭示了CDK功能的新方面，为进一步研究这些关键蛋白激酶提供了方向。
Cyclin-dependent kinases (CDKs) are the catalytic subunits of a family of mammalian heterodimeric serine/threonine kinases that play critical roles in the control of cell-cycle progression, transcription, and neuronal functions. However, the functions, substrates, and regulation of many CDKs are poorly understood. To systematically investigate these features of CDKs, we conducted a proteomic analysis of the CDK family and identified their associated protein complexes in two different cell lines using a modified SAINT (Significance Analysis of INTeractome) method. The mass spectrometry data were deposited to ProteomeXchange with identifier PXD000593 and DOI 10.6019/PXD000593. We identified 753 high-confidence candidate interaction proteins (HCIPs) in HEK293T cells and 352 HCIPs in MCF10A cells. We subsequently focused on a neuron-specific CDK, CDK5, and uncovered two novel CDK5-binding partners, KIAA0528 and fibroblast growth factor (acidic) intracellular binding protein (FIBP), in non-neuronal cells. We showed that these three proteins form a stable complex, with KIAA0528 and FIBP being required for the assembly and stability of the complex. Furthermore, CDK5-, KIAA0528-, or FIBP-depleted breast cancer cells displayed impaired proliferation and decreased migration, suggesting that this complex is required for cell growth and migration in non-neural cells. Our study uncovers new aspects of CDK functions, which provide direction for further investigation of these critical protein kinases.