蛋白向细胞质膜的迁移定位导致细胞坏死.docxVIP

AbstractAbstract Abstract Abstract Mixed lineage kinase domain一1ike protein(Mlkl)was identified as downstream of Receptor interaction protein 3(Rip3)in Tumor Necrosis Factor alpha(TNF a)induced necrosis．However，the exact function of Mlkl in necrosis is largely unknown． By reconstituting exogenous Mlkl in Mlkl—K0 cell，we found the integrity of N-terminus domain of Mlkl was required for it normal function． 01igomerization of mlkl was essential for necrosis，as artificially forced Mlkl oligomers by fusing hormone—binding domain(HBD*)triggered necrosis． Notably， artificially forced oligomers of N—Terminal domains(ND-mlkl)but not C—terminal kinase 1 ike domain caused necrosis． Further deletion mutations showed that the four—G—helix bundle of Mlkl(1—130 amino acids)iS sufficient for necrosiS．Both TNF induced endogenous Mlkl oligomers in wild type cell or HBD水mediated ND-mlkl oligomers were tetramers．Both of the homo—ol igomers were observed to translocate to 1 ipid rafts on cel lular plasma membrane via immunofluorescence confocal microscopy．The membrane translocated signal sequence in in the junction of the first and second Q-helices．Plasma membrane translocation of Mlkl ND—mlkl leads to increased cellular plasma sodium concentration，indicated by CoroNa Green signal strength largely enhanced detected via immunofluorescence confocal microscopy． Depletion of sodium in cell culture medium inhibited necrosiS．A11 of the above phenomena were not observed in apoptos is．Thus，we came to conclusion that，Mlkl oligomerization leads to translocation to 1ipid rafts on cellular plasma membrane，after that，Mlkl complex acts either by itself or via other proteins to increase sodium influx，which increased cellular osmotic pressure and kept cell intaking more water， eventual ly leaded to plasma rupture． Keywords：Mlkl：necrosis：incresed plasmic sodium concentration： Rip3：translocation to plasma membrane． Ⅱ 万方数据 目录目录 目录 目录 摘要 ． ． ．I Abst伯ct ． ． ．．．．1 I 目录II l 第一章 前言．．．．．． ．．． ．．． ．．．