title: Cys92, Cys101, Cys197, and Cys203 are crucial residues for coordinating the iron-sulfur cluster of RhdA from Acidithiobacillus ferrooxidans.
abstract: 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.
authors.affiliation: Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, 410083 Changsha, Hunan, People's Republic of China. dairenjie1009@163.com
authors.forename: Yunjie
authors.initials: Y
authors.lastname: Dai
authors.forename: Jianshe
authors.initials: J
authors.lastname: Liu
authors.forename: Chunli
authors.initials: C
authors.lastname: Zheng
authors.forename: Anna
authors.initials: A
authors.lastname: Wu
authors.forename: Jia
authors.initials: J
authors.lastname: Zeng
authors.forename: Guanzhou
authors.initials: G
authors.lastname: Qiu
mesh_terms: D042763:Acidithiobacillus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007501:Iron / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D016415:Sequence Alignment; D013455:Sulfur / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D013455:Sulfur; D007501:Iron; D003545:Cysteine