低表达线粒体蛋白ISCA2对线粒体功能影响的机制研究  

Elucidation of Roles of Protein ISCA2 in the Control of Mitochondrial Function

作　　者：杨玲 刘若兰[1] 牛欢[1] 胡立 李艳纯[1] 王伟龙 李江辉[1] 谭国强[1] 吕建新[1] Yang Ling;Liu Ruolan;Niu Huan;Hu Li;Li Yanchun;Wang Weilong;Li Jianghui;Tan Guoqiang;Lu Jianxin(School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学检验医学院生命科学学院,温州325035

出　　处：《中国细胞生物学学报》2018年第7期1120-1129,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81671124;81500440)资助的课题~~

摘　　要：该文探讨了ISCA2蛋白低表达对细胞内铁硫蛋白和能量代谢的影响。在HeLa细胞内将ISCA2基因敲低,通过免疫印迹、顺乌头酸酶胶内酶活性分析法分析线粒体内外铁硫蛋白水平和活性改变;用紫外–可见分光光度法检测细胞线粒体内氧化磷酸化复合体活性;用海马能量分析仪分析细胞内能量代谢的改变。结果表明,ISCA2蛋白低表达后,氧化磷酸化复合体中各铁硫蛋白亚基都出现不同程度的下调,且对于线粒体[4Fe-4S]型铁硫蛋白亚基影响显著,但对线粒体[2Fe-2S]型铁硫蛋白亚基以及胞质[4Fe-4S]型铁硫蛋白亚基影响较小。同时,ISCA2蛋白低表达后对线粒体复合体活性和线粒体能量代谢影响显著,细胞有氧呼吸降低,细胞外乳酸含量增加。这些结果表明,ISCA2蛋白低表达后抑制铁硫簇的组装,导致铁硫蛋白功能障碍,影响线粒体复合体活性和氧化磷酸化系统,使得细胞能量代谢紊乱。This article explored the effect of low expression of ISCA2 on intracellular iron-sulfur protein and energy metabolism. Firstly, we conducted the RNA-mediated knockdown of ISCA2 gene. And then, we detected the activity changes of iron-sulfur proteins inside and outside the mitochondria by Western blot and in-gel activity assay. Furthermore, the Seahorse Bioscience XF96 Extracellular Flux Analyzer was used to analyze the changes in intracellular energy metabolism. The results showed that the iron-sulfur subunits in respiratory chain complexes were decreased to some extent after low expression of ISCA2 protein. And a substantial decrease of mitochondrial [4 Fe-4 S] containing proteins was observed by Western blot after low expression of ISCA2 protein, while a slight reduction in the cytoplasmic [4 Fe-4 S] enzyme and mitochondrial [2 Fe-2 S] containing proteins was observed. In addition, mitochondrial complex activity and energy metabolism were damaged after the decrease of ISCA2 protein.

关 键 词：铁硫簇 铁硫蛋白 能量代谢 

分 类 号：R363[医药卫生—病理学]