具有泛素及nedd8调节功能的病源蛋白在巨噬细胞中的功能探究.ppt

具有泛素及Nedd8调节功能的病源蛋白在巨噬细胞中的功能探究 陈龙 北京大学 生命科学学院 Long Chen School of Life-Science， Peking University 11/26/2011 目 录 Background 1 Results 2 Conclusion and future work 3 Acknowledgements 4 Backgroud Ubiquitin(泛素) and Nedd8 Ubiquitin ——2004 Noble Price Nedd8 is similar to Ubiquitin(cell cycle（细胞周期） progression and cytoskeletal（细胞骨架） regulation) DUB (How Ub works（泛素化原理）,Modified from Jixin Cui) Background Cif and family protein(especially CHBP) (Q. Yao et al., PNAS U.S.A. 106, 3716 (2009)) Bacterial Effector Cif——Enteropathogenic E. coli（大肠杆菌） CHBP——Burkholderia pseudomallei（鼻疽杆菌） Background Glutamine（谷氨酸） Deamidation（脱氨基作用） and Dysfunction of Ubiquitin/NEDD8 Induced by CHBP and Cif Difference between Cif and CHBP Ubiquitin Nedd8 Background Macrophage（巨噬细胞） (Macrophage,From Wikipedia) (Immune System（免疫系统）, from Nature) Questions we want to know How could we deliver the protein into the macrophage How do the different protein effect on the macrophage The reason for the influence on the macrophage First Second Finally Results By osmotic（渗透压） lysis, the protein didnt show great influence on the macrophage.The osmotic lysis is not sensitive enough for this experiment. Yogendra Singh, et at, JBC, 1989 (ATP assay for osmotic lysis on P80 cell, 24h) Results PA(Protective Antigen)-LF(Lethal Factor，致死因子)n system. Preparation for LFn-CHBP, LFn-CHBP C/A, LFn-Cif, LFn-Cif C/A, LFn and PA. LFn CHBP C/S LFn Cif PA LFn CHBP LFn Cif C/A PA-LF/EF system from anthrax bacteria（炭疽杆菌） , from google Results LFn-CHBP leads the macrophage to cell death while LFn-Cif doesnt effect the macrophage seriously. Mock CHBP Cif Macrophage P80 Results LFn-CHBP leads the macrophage to cell death while LFn-Cif doesnt effect the macrophage seriously. ATP assay(mock, PA+LFn-CHBP and LFn-Cif, 24h.) Results The poor influence of LFn-Cif is mainly due to the failure to enter the macrophage. Hela（海拉细胞） Mock