Protein microarray analysis identifies human cellular prion protein interactors
Protein microarray analysis identifies human cellular prion protein interactors
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DOI：
10.1111/j.1365-2990.2008.00947.x
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发表时间：
2009-02-01
影响因子：
5
通讯作者：
Tabunoki, H.
中科院分区：
文献类型：
--
作者：
Satoh, J.;Obayashi, S.;Tabunoki, H.
目的：通过对蛋白质芯片的分析，研究PrPC相互作用蛋白(PrPIPs)，以深入了解PrPC的功能。方法：我们用编码23-231个氨基酸残基的重组人PrPC序列探测5000个人类蛋白质，鉴定出47个新的PrPIPs，命名为PR209。结果：在47个PrPIP中，绝大多数被注释为参与核酸识别的蛋白质。免疫共沉淀和瞬时表达系统中的细胞成像证实了PR209与神经元PrPIP如FAM64A、HOXA1、PLK3和MPG的相互作用。然而，这种相互作用并没有产生抗蛋白酶K的蛋白。KeyMolnet是一种生物信息学工具，用于分析精选知识库中的分子相互作用，它揭示了PrPC和PrPIPs的复杂分子网络与AKT、JNK和MAPK信号通路有着重要的关系。结论：蛋白质芯片是系统筛选和全面分析人类PrPC相互作用组的有用工具。由于PrPC及其相互作用分子网络涉及对细胞生存、分化、增殖和凋亡的调控至关重要的信号通路，这些观察结果提出了一个逻辑假设，即PrPC相互作用组的调节失调可能会导致普恩疾病的广泛神经退行性变。
Aims: To obtain an insight into the function of cellular prion protein (PrPC), we studied PrPC-interacting proteins (PrPIPs) by analysing a protein microarray. Methods: We identified 47 novel PrPIPs by probing an array of 5000 human proteins with recombinant human PrPC spanning amino acid residues 23-231 named PR209. Results: The great majority of 47 PrPIPs were annotated as proteins involved in the recognition of nucleic acids. Coimmunoprecipitation and cell imaging in a transient expression system validated the interaction of PR209 with neuronal PrPIPs, such as FAM64A, HOXA1, PLK3 and MPG. However, the interaction did not generate proteinase K-resistant proteins. KeyMolnet, a bioinformatics tool for analysing molecular interaction on the curated knowledge database, revealed that the complex molecular network of PrPC and PrPIPs has a significant relationship with AKT, JNK and MAPK signalling pathways. Conclusions: Protein microarray is a useful tool for systematic screening and comprehensive profiling of the human PrPC interactome. Because the network of PrPC and interactors involves signalling pathways essential for regulation of cell survival, differentiation, proliferation and apoptosis, these observations suggest a logical hypothesis that dysregulation of the PrPC interactome might induce extensive neurodegeneration in prion diseases.