Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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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)
  • Published : 2008.10.31
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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

References

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