国家重点基础研究发展计划(973)(2006CB500700), 国家高科技研究发展计划(863)(2006AA02A408)和国家自然科学基金(30570638, 30770735, 30971035, 30900469)资助项目
This work was supported by grants from National Basic Research Program of China(2006CB500700), Hi-Tech Research and Development Program of China(2006AA02A408) and The National Natural Science Foundation of China (30570638, 30770735, 30971035, 30900469)
帕金森病(Parkinson's disease,PD)是常见的神经系统变性疾病.分子遗传学研究发现,突变的Parkin蛋白及PINK1蛋白均参与了帕金森病的致病过程,但二者之间是否存在相互作用以及是否能够相互调节仍不十分清楚.为明确生理状态下Parkin蛋白与PINK1蛋白之间的相互作用,首先运用蛋白体外结合实验(GST pull-down)技术及免疫共沉淀技术证实了Parkin与PINK1在体外及体内均可相互结合.进一步构建PINK1的不同截短型,运用GST pull-down技术验证了PINK1与Parkin相互结合的区段为PINK1的蛋白激酶结构域.免疫细胞化学实验也证实Parkin与PINK1蛋白在细胞中存在共定位.进一步运用免疫共沉淀技术证实Parkin可减少PINK1通过泛素蛋白酶体系统(ubiquitin proteasome system,UPS)的降解,从而稳定PINK1.PINK1可增加Parkin通过UPS的降解,从而减少Parkin的水平,降低其稳定性.这些结果提示,帕金森病相关蛋白Parkin与PINK1能够直接结合,二者通过泛素蛋白酶体降解系统相互调节,可能协同作用参与了帕金森病的致病过程.
Parkinson disease (PD) is the common movement disorder. Along with the development of molecular genetics, genes related to the familial forms of Parkinson's disease such as Parkin (PARK2) and phosphatase and tensin homologue deleted on chromosome ten (PTEN)-induced putative kinase 1——PINK1 (PARK6) have been identified. GST pull-down assays were performed to identify which domain of PINK1 interacted with Parkin, and the result showed that Parkin directly interacted with PINK1, and the PINK1 kinase domain interacted with Parkin. Based on these results, then the co-immunoprecipitation technique was used to investigate the possible interaction between Parkin and PINK1 in 293A cells. The results indicated that Parkin stabilized PINK1 by interfering with its degradation via the ubiquitin-mediated proteasomal pathway, and PINK1 decreased level of Parkin by promoting its degradation via the ubiquitin-mediated proteasomal pathway. The results confirmed that Parkin directly interacted with PINK1, and they regulated each other via the ubiquitin-mediated proteasomal pathway.
王雪晶,郭纪锋,江泓,沈璐,唐北沙.帕金森病相关蛋白Parkin 与PINK1的相互作用研究[J].生物化学与生物物理进展,2010,37(9):983-987
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