NPC2蛋白与PARK9 F1区域蛋白互作研究

发布时间：2018-06-19 22:02

本文选题：C型尼曼-皮克病 + NPC2　；参考：《杭州师范大学》2015年硕士论文

【摘要】：C型尼曼-皮克病(Niemann-Pick type C,NPC)是一组常染色体隐性遗传病。目前认为,NPC是一种独特的细胞内、外源性胆固醇和多种脂质转运障碍所致的变性病。5%的NPC病例源于结合胆固醇的可溶性溶酶体蛋白NPC2基因突变。PARK9基因是常染色体隐性遗传青壮年帕金森综合征(Kufor-Rakeb综合征,简称KBS综合征)的致病基因,又称ATP13A2基因。本课题组前期研究数据表明,NPC2与PARK9或许存在互作的关系。编码的PARK9蛋白可能参与了调控细胞内物质转运这项活动。本课题研究主要采用计算机模拟、体外重组的技术,体外表达NPC2蛋白和PARK9基因片段,且成功获得相应蛋白;银染与GST pull-down方法,表明NPC2和PARK9存在着相互作用的关系;同时用Dot blot实验初步研究了二者间相互作用的量子化关系,即二者浓度接近1:1时,NPC2与PARK9 F1体积在1:4-1:1之间结合较好。该课题通过对NPC2与PARK9间的互作研究,以期揭示PARK9蛋白可通过与NPC2的结合参与调控溶酶体内胆固醇转运。
[Abstract]:Niemann-Pick type is a group of autosomal recessive diseases. At present, it is believed that the NPC is a unique kind of intracellular, 5% of NPC cases caused by exogenous cholesterol and various lipid transport disorders were caused by mutation of soluble lysosomal protein NPC2 gene. PARK9 gene was autosomal recessive young and adult Parkinson's syndrome and Kufor-Rakeb syndrome. The pathogenicity gene of KBS syndrome, also known as ATP 13A2 gene. The previous study data suggested that there may be interaction between NPC2 and PARK9. The encoded PARK 9 protein may be involved in regulating the intracellular transport of substances. In this study, NPC2 and PARK9 gene fragments were expressed by computer simulation and recombinant in vitro, and the corresponding proteins were obtained successfully, and the interaction between NPC2 and PARK9 was demonstrated by silver staining and GST pull-down method. At the same time, the quantization relationship of the interaction between them is studied by Dot blot experiment. The results show that the volume of NPC2 and PARK9F1 is better combined with the volume of 1: 4-1: 1 when the concentration of NPC2 and PARK9F1 is close to 1:1. Through the study of the interaction between NPC2 and PARK9, we hope that PARK9 protein can regulate cholesterol transport by binding with NPC2.
【学位授予单位】：杭州师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R596.1

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