半胱氨酸92、半胱氨酸101、半胱氨酸197和半胱氨酸203是用于配位嗜酸氧化亚铁硫杆菌RhdA的铁硫簇的关键残基。
Cys92, Cys101, Cys197, and Cys203 are crucial residues for coordinating the iron-sulfur cluster of RhdA from Acidithiobacillus ferrooxidans.
作者信息
Dai Yunjie, Liu Jianshe, Zheng Chunli, Wu Anna, Zeng Jia, Qiu Guanzhou
机构信息
Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, 410083 Changsha, Hunan, People's Republic of China.
出版信息
Curr Microbiol. 2009 Nov;59(5):559-64. doi: 10.1007/s00284-009-9476-x.
By proteomic analysis, we found a rhodanese-like protein(RhdA) from Acidithiobacillus ferrooxidans ATCC 23270 whose C-terminal contained a cysteine motif (Cys-XX-Trp-XX-Cys), known to bind iron-sulfur clusters. But so far, there were no articles to confirm the existence of iron-sulfur cluster in RhdA. In this study, RhdA gene from A. ferrooxidans ATCC 23270 was cloned and expressed in Escherichia coli, the protein was purified by one-step affinity chromatography to homogeneity. The UV-Vis scanning and EPR spectra results indicated that the wild-type proteins contained an iron-sulfur cluster. Site-directed mutagenesis results revealed that the four cysteines Cys92, Cys101, Cys197, and Cys203 were crucial residues for iron-sulfur cluster binding.
通过蛋白质组学分析，我们从嗜酸氧化亚铁硫杆菌ATCC 23270中发现了一种类硫氰酸酶蛋白（RhdA），其C端含有一个半胱氨酸基序（Cys-XX-Trp-XX-Cys），已知该基序可结合铁硫簇。但到目前为止，尚无文章证实RhdA中存在铁硫簇。在本研究中，克隆了嗜酸氧化亚铁硫杆菌ATCC 23270的RhdA基因并在大肠杆菌中表达，通过一步亲和层析将该蛋白纯化至均一。紫外可见扫描和电子顺磁共振光谱结果表明，野生型蛋白含有一个铁硫簇。定点诱变结果显示，四个半胱氨酸Cys92、Cys101、Cys197和Cys203是铁硫簇结合的关键残基。
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