Tn细胞凋亡抑制蛋白(TnIAP)在粉纹夜蛾细胞中的表达和活性研究  

Expression of TnIAP in Trichoplusia ni Cells and Its Biological Activity

作　　者：廖文韬[1] 杨扬[1] 吴祥甫[1] 

机构地区：[1]中国科学院上海生命科学研究院生物化学与细胞生物学研究所,上海200031

出　　处：《生物化学与生物物理进展》2002年第5期741-744,共4页Progress In Biochemistry and Biophysics

摘　　要：在粉纹夜蛾 (Trichoplusiani)细胞Tn 5B1 4中 ,高效表达了来自粉纹夜蛾细胞Tn 5B1 4能够抑制细胞凋亡的TnIAP蛋白 .聚丙烯酰胺凝胶电泳和免疫印迹分析表明 ,表达的重组TnIAP只有少部分是可溶性蛋白 ,大部分以不溶的形式存在 .这一结果与以往在昆虫细胞中往往表达出可溶性蛋白不同 .活性实验表明 ,可溶的重组TnIAP能够直接抑制caspase 9酶解Ac LEHD AFC的活性 ,也能抑制caspase 9激活HEK2 93细胞抽提物酶解Ac DEVD AFC的活性 .结果进一步证明 。TnIAP, which can inhibit apoptosis and was cloned from Tri-5B1-4 cells, was highly expressed in Tn-5B1-4 cells. However, SDS-PAGE and Western blot analysis indicated that only small part of recombinant protein was soluble, which was different from the record that most recombinant protein expressed in insect cells were soluble. Activity assays demonstrated that recombinant soluble TnIAP could inhibit the cleavage of Ac-LEHD-AFC by caspase-9 and the activation of HEK293 cytosolic extracts by caspase-9 from cleaving Ac-DEVD-AFC. This result gave the further evidence that the molecular mechanism of apoptosis is evolutionarily conserved through insects to mammals.

关 键 词：Tn细胞凋亡抑制蛋白 TnIAP 粉纹夜蛾 细胞凋亡 表达 活性 

分 类 号：Q255[生物学—细胞生物学] Q26