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为了探讨蛋白激酶CK2与PrP是否存在分子间相互作用,利用RT-PCR方法从人源细胞系SH-SY5YcDNA中分别扩增出蛋白激酶CK2α和CK2β基因,并在E.coli中表达了融合蛋白HIS-CK2α和GST-HIS-CK2β。利用pull-down及免疫共沉淀实验检测PrP与CK2的分子间相互作用。结果显示,重组PrP可与CK2α出现特异性结合,但不与CK2β发生反应。天然状态下脑组织中的CK2与PrPC也发生相互作用。PrP与CK2α亚基相互作用的区域位于PrP的C-端90～231位氨基酸。本研究从分子水平上提供了人重组和天然CK2和PrP蛋白相互作用的实验依据,为深入探讨CK2与朊蛋白作用的生物学意义和在朊病毒病发病过程中的作用提供了科学线索。
In order to investigate whether there is intermolecular interaction between protein kinase CK2 and PrP, protein kinase CK2α and CK2β genes were amplified from human cell line SH-SY5Y by RT-PCR and expressed in E.coli HIS -CK2α and GST-HIS-CK2β. Pull-down and co-immunoprecipitation assays were used to detect the intermolecular interactions between PrP and CK2. The results showed that recombinant PrP with CK2α specific binding, but does not react with CK2β. The natural state of brain tissue CK2 and PrPC also interact. The region of PrP that interacts with the CK2 alpha subunit is located at the C-terminal amino acid 90 to 231 of PrP. This study provides experimental evidence for the interaction between human recombinant and native CK2 and PrP proteins at the molecular level and provides scientific clues for further exploring the biological significance of CK2 and prion protein’s role in the pathogenesis of prion diseases.