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KMID : 0545120080180101672
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Journal of Microbiology and Biotechnology
2008 Volume.18 No. 10 p.1672 ~ p.1677
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Expression, Purification, and Characterization of Iron-Sulfur Cluster Assembly Regulator IscR from Acidithiobacillus ferrooxidans
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Zeng Jia
Zhang Ke
Liu Jianshe
Qiu Guanzhou
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Abstract
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IscR (iron-sulfur cluster regulator) has been reported to be a repressor of the iscRSUA operon, and in vitro transcription reactions have revealed that IscR has a repressive effect on the iscR promoter in the case of [Fe2S2] cluster loading. In the present study, the iscR gene from A. ferrooxidans ATCC 23270 was cloned and successfully expressed in Escherichia coli, and then purified by one-step affinity chromatography to homogeneity. The molecular mass of the IscR was 18 kDa by SDS-PAGE. The optical and EPR spectra results for the recombinant IscR confirmed that an iron-sulfur cluster was correctly inserted into the active site of the protein. However, no [Fe2S2] cluster was assembled in apoIscR with ferrous iron and sulfide in vitro. Therefore, the [Fe2S2] cluster assembly in IscR in vivo would appear to require scaffold proteins and follow the Isc “AUS” pathway.
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KEYWORD
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Acidithiobacillus ferrooxidans, His-tag, ironsulfur cluster, assembly, regulator
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