eIF4A1与TrkA相互作用后抑制TrkA的泛素化（英文）
发布时间：2021-04-13 08:23
神经生长因子（NGF）结合细胞表面受体p75NTR （p75神经营养素受体）和TrkA （酪氨酸蛋白激酶A）后介导了细胞分化、细胞生存、凋亡、增殖和侵袭等多个重要的生理病理过程. TrKA能与细胞内多个蛋白质相互作用,但是由于NGF信号通路的复杂性,现在仍有必要发现与之相互作用的蛋白质以更准确地了解NGF信号通路.本研究中我们通过酵母双杂交的方法筛选到了一个新的与TrKA相互作用的蛋白质——真核生物翻译起始因子4A1 （eIF4A1）,然后通过谷胱甘肽巯基转移酶融合蛋白沉降实验（GST-pull-down）和免疫共沉淀实验（Co-IP）证明了TrkA和eIF4A1的相互作用.此外NGF能够增强TrkA和eIF4A1的相互作用.在鉴定相互作用位点实验中,我们发现eIF4A1的氨基端结构域和TrkA的TK结构域参与了相互作用. TrkA和e IF4A1共定位在细胞膜上. NGF能够引起TrkA与泛素蛋白63位的赖氨酸连接,而eIF4A1与TrkA相互作用后能够抑制TrkA与泛素蛋白63位的赖氨酸连接.综上,得出结论 e IF4A1通过与TrkA相互作用抑制其泛素化调控NGF信号通路.
【文章来源】：生物化学与生物物理进展. 2019,46(08)北大核心SCICSCD
【文章页数】：11 页
【文章目录】：
1 Methods
1.1 Yeast two hybrid screening
1.2 Cell culture, transfection and NGF stimulation
1.3 Plasmids, antibodies and reagents
1.4 GST pull-down assay
1.5 Co-immunoprecipitation and immunoblotting
1.6 Immunofluorescence assay
1.7 Cytoplasmic and nuclear extraction
2 Results
2.1 Identification of the novel association of TrkA and eIF4A1
2.2 eIF4A1 binds to TrkA-ICD in vivo and in vitro
2.3 Mapping the interacting domain between TrkA and eIF4A1
2.4 Membrane co-localization of TrkA and eIF4A1
2.5 NGF stimulated TrkA Lys 63-linked polyubiquitination and eIF4A1 repressed TrkApolyubiquitination
3 Discussion
本文编号：3134958
【文章来源】：生物化学与生物物理进展. 2019,46(08)北大核心SCICSCD
【文章页数】：11 页
【文章目录】：
1 Methods
1.1 Yeast two hybrid screening
1.2 Cell culture, transfection and NGF stimulation
1.3 Plasmids, antibodies and reagents
1.4 GST pull-down assay
1.5 Co-immunoprecipitation and immunoblotting
1.6 Immunofluorescence assay
1.7 Cytoplasmic and nuclear extraction
2 Results
2.1 Identification of the novel association of TrkA and eIF4A1
2.2 eIF4A1 binds to TrkA-ICD in vivo and in vitro
2.3 Mapping the interacting domain between TrkA and eIF4A1
2.4 Membrane co-localization of TrkA and eIF4A1
2.5 NGF stimulated TrkA Lys 63-linked polyubiquitination and eIF4A1 repressed TrkApolyubiquitination
3 Discussion
本文编号：3134958
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/3134958.html
最近更新
教材专著
