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Elucidation of Roles of Protein ISCA2 in the Control of Mitochondrial Function
Yang Ling, Liu Ruolan, Niu Huan, Hu Li, Li Yanchun, Wang Weilong, Li Jianghui, Tan Guoqiang, Lü Jianxin*
School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China
Abstract: 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 [4Fe-4S] containing proteins was observed by Western blot after low expression of ISCA2 protein， while a slight reduction in the cytoplasmic [4Fe-4S] enzyme and mitochondrial [2Fe-2S] containing proteins was observed. In addition， mitochondrial complex activity and energy metabolism were damaged after the decrease of ISCA2 protein.
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