rVvhA功能模序膜成孔的预测及细胞毒作用机制分析  

The Prediction and Cytotoxicity Mechanism Analysis on the Functional Motif of the Membrance Pore-forming in rVvhA

作　　者：刘艳飞[1] 陈建林[1] 许玲[1] 楼永良[1] 

机构地区：[1]温州医学院检验医学院,温州325035

出　　处：《中国细胞生物学学报》2012年第6期527-537,共11页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金(No.Y2090468)资助项目~~

摘　　要：利用生物信息学预测rVvhA的141-335位氨基酸片段有膜成孔模序。基因克隆表达得到95%以上纯度的rMpf,电子透射电镜观察其能够抑制Hela细胞生长且呈剂量依赖性,即0.8,1.6,2.4μg/mL rMpf作用8 h后,细胞和线粒体形态均发生凋亡和坏死改变,细胞内活性氧产生明显,线粒体膜电位下降,mPTP荧光检测膜通道孔活性增强。以上结果表明,rMpf具有诱导Hela细胞损伤的生物学活性,可通过改变膜通透性引起细胞凋亡。It was predicted that the area between the sites 141-335 of rVvhA amino acid sequence has the membrane pore-forming motif through the methods of bioinformatic. Using methods of geng-cloning, purification and renaturation, the rMpf proteins with purity of more than 95% was obtained. It can inhibit the growth of Hela cells and the effect was depended on doses, 0.8, 1.6, 2.4 μg/mL rMpf was used respectively for 8 hours, by transmission electron microscopy it was found that the cells show characteristic of apoptosis and the structure of mitochondrion was altered; intra-cellular ROS production was increased obvioulsy; mitochondrial membrane potential was lowered; mitochondrial permeability transition pore activity was increased as revealed by mPTP＇s fluorescence inspection. The above data show that rMpf plays a role in inducing Hela cells＇ damage, and leads to cell apoptosis via influencing cell membrane permeability.

关 键 词：溶细胞素 生物信息学 凋亡 线粒体 活性氧 

分 类 号：R378[医药卫生—病原生物学]