在粉纹夜蛾(Trichoplusia ni)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HEK293细胞抽提物酶解Ac-DEVD-AFC的活性.结果进一步证明,昆虫和哺乳动物的细胞凋亡分子机制在进化上是极为保守的.
TnIAP, which can inhibit apoptosis and was cloned from Tn-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.
廖文韬,杨扬,吴祥甫. 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[J].生物化学与生物物理进展,2002,29(5):741-744
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