ORP8与SPAG5相互作用并影响细胞周期  

Interaction of ORP8 with SPAG5 and Their Effects on Cell Cycle

作　　者：罗玮[1] 周天鸿[1] 闫道广[1] 

机构地区：[1]暨南大学生命科学技术学院,广东广州510632

出　　处：《现代生物医学进展》2011年第14期2601-2604,2623,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金(30971104)

摘　　要：目的:筛选与氧化固醇结合蛋白相关蛋白8(Oyxterol binding protein related protein 8,ORP8)相互作用的蛋白质,为揭示ORP8在细胞中的功能及其机制提供线索,并根据相互作用蛋白质初步探索ORP8的功能。方法:构建重组诱饵质粒pGBKT7-ORP8m,利用GAL4酵母双杂交系统筛选人Universal cDNA文库,筛选出与ORP8相互作用的蛋白质,通过GSTPull-down以及免疫共沉淀(Co-immunoprecipitation,Co-IP)验证蛋白质相互作用,并通过流式细胞仪检测过表达ORP8对HepG2细胞周期的影响。结果:酵母双杂交筛选得到了精子相关抗原5(Homo sapiens sperm associated antigen 5,SPAG5),体外GSTpull-down和Co-IP实验确证了ORP8-SPAG5的相互作用,流式细胞术显示ORP8过表达后与空载体相比,人肝癌细胞系HepG2的S期细胞数增加,G1期细胞数减少。结论:SPAG5是与ORP8相互作用的蛋白质,ORP8过表达影响人肝癌细胞系HepG2的细胞周期,可能通过SPAG5起作用。Objective： To investigate the function of oxyterol binding protein related protein 8（ORP8）and to provide clues for better understanding its function and mechanism.Methods： Yeast two-hybrid system screening human universal cDNA library was used to obtained proteins interacting with ORP8.Putative interactions were further confirmed by GST Pull-down and Co-IP experiments.The effects of ORP8 on cell cycle were detected by flow cytometry in HepG2 cells.Results： ORP8 interacted with SPAG5 in two-hybrid screen using ORP8 as bait.ORP8-SAPG5 interaction was further confirmed in GST Pull-down and Co-IP experiments.Transient over-expression of ORP8 in HepG2 cells decreased cell population of G0/G1-phase and increased S-phase compared with that in control.Conclusion： ORP8 physically interact with SPAG5 and influence the cell cycle of HepG2 cell lines,probably through a mechanism in-volving SPAG5.ORP8-SPAG5 interaction provides clues to further explore the function of ORP8 and its mechanism.

关 键 词：ORP8 酵母双杂交 SPAG5 蛋白质相互作用 细胞周期 

分 类 号：Q548.1[生物学—生物化学] Q78