摘要
目的泛素化是机体重要的蛋白质的修饰方式,人环指蛋白(ring finger protein,RNF)11是泛素连接酶家族成员之一,可能在肿瘤形成过程中发挥重要的作用。肿瘤坏死因子α诱导蛋白3相互作用蛋白1(tumor necrosis factorα-induced pro-tein 3-interacting protein 1,TNIP1)是重要的核因子κB(nuclear factor kB,NF-κB)调控蛋白,可通过多种途径调控NF-κB的表达及细胞凋亡。文中研究RNF11下游功能及其与TNIP1之间的相互作用,明确RNF11的亚细胞定位。方法用PCR等方法构建人NFκB调控蛋白TNIP1全长及缺失不同位点的突变体,并观测其在293T细胞中的表达。用免疫共沉淀(co-immunoprecipita-tion,co-IP)方法鉴定其与人RNF11的相互作用,并确定其结合位点,用共聚焦显微镜观测其亚细胞定位。结果成功构建了人TNIP1的全长及其缺失不同位点的突变体并使其在293T细胞中表达。证实RNF11与TNIP1存在相互作用并确定其相互结合位点位于TNIP1的第311至535位氨基酸,。确定RNF11的亚细胞定位于细胞质。结论人RNF11可与TNIP1在细胞内相互结合,其主要的结合位点位于311至535位氨基酸。RNF11主要定位于细胞质,可能参与细胞内蛋白的降解过程。
Objective The purpose of this study was to investigate the sub-function of RNF11 and its interaction with TNIP1, and to identify the subcellular location of RNF11. Methods We constructed the expression vector of human NF-κB inhibitors TNIP1 and its mutation by PCR, observed its expression in 293T cells, and identified the site of its interaction with RNFll by coimmunoprecipitation as well as the subcellular location of RNF11 by co-focal microscopy. Results We successfully constructed the eukaryotic expression vector of TNIP1 and its mutation, which expressed in 293T cells. RNF11 and TNIP1 interacted between each other, and their binding site was between 311 and 535 aa, and the subcellular location of RNF11 was mainly in the endochylema. Conclusion Human RNF11 interacts with NF-κB inhibitor TNIP1 intracellularly between 311 and 535 aa, localized mainly in the endochylema, and maybe involved in the process of protein degradation.
出处
《医学研究生学报》
CAS
北大核心
2012年第6期597-600,共4页
Journal of Medical Postgraduates
