Global Analysis of Human Prion Protein Interactors by Protein Microarray

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Aims : To obtain an insight into the function of cellular prion protein(PrPC), we studied PrPC-interacting proteins(PrPIPs) by analyzing a protein microarray. Methods : We idenitified 47 novel PrPIPs by probing the array of 5,000 human proteins with recombinant human PrPC spanning amino acid residues 23-231 named PR209. Results : The great majority of 47 PrPIPs were annotated as the 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 analyzing molecular interaction on the curated knowledge database, revealed that the complex molecular network of PrPC and PrPIPs has the significant relationship with AKT, JNK and MAPK signaling 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 signaling pathways essential for regulation of cell survival, differentiation, proliferation and apoptosis, these observations propose a logical hypothesis that dysregulation of the PrPC interactome might induce extensive neurodegeneration in prion diseases.