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Expression, Purification, and Characterization of Iron-Sulfur Cluster Assembly Regulator IscR from Acidithiobacillus ferrooxidans  

Zeng, Jia (Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University)
Zhang, Ke (Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University)
Liu, Jianshe (Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University)
Qiu, Guanzhou (Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University)
Publication Information
Journal of Microbiology and Biotechnology / v.18, no.10, 2008 , pp. 1672-1677 More about this Journal
Abstract
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 [$Fe_{2}S_{2}$] 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 [$Fe_{2}S_{2}$] cluster was assembled in apoIscR with ferrous iron and sulfide in vitro. Therefore, the [$Fe_{2}S_{2}$] cluster assembly in IscR in vivo would appear to require scaffold proteins and follow the Isc "AUS" pathway.
Keywords
Acidithiobacillus ferrooxidans; His-tag; iron-sulfur cluster; assembly; regulator;
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