Abstract
The [2Fe-2S] cluster containing ferredoxin has attracted much attention in recent years. Genetic analyses show that it has an essential role in the maturation of various iron-sulfur (Fe-S) proteins and functions as a component of the complex machinery responsible for the biogenesis of Fe-S clusters. The gene of ferredoxin from A. ferrooxidans ATCC 23270 was cloned, successfully expressed in Escherichia coli, and purified by one-step affinity chromatography to homogeneity. The MALDI-TOF MS and spectra results of the recombinant protein confirmed that the iron-sulfur cluster was correctly inserted into the active site of the protein. Site-directed mutagenesis results revealed that Cys42, Cys48, Cys51, and Cys87 were ligating with the [Fe(2)S(2)] cluster of the protein.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Acidithiobacillus thiooxidans / enzymology*
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Acidithiobacillus thiooxidans / genetics
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Acidithiobacillus thiooxidans / metabolism
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Amino Acid Sequence
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Bacterial Proteins / chemistry
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Bacterial Proteins / genetics
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Bacterial Proteins / isolation & purification
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Bacterial Proteins / metabolism
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Chromatography, Affinity
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Cloning, Molecular
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Escherichia coli / enzymology*
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Escherichia coli / genetics
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Ferredoxins* / chemistry
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Ferredoxins* / genetics
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Ferredoxins* / isolation & purification
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Ferredoxins* / metabolism
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Iron-Sulfur Proteins* / chemistry
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Iron-Sulfur Proteins* / genetics
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Iron-Sulfur Proteins* / isolation & purification
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Iron-Sulfur Proteins* / metabolism
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Molecular Sequence Data
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Mutagenesis, Site-Directed
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Substances
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Bacterial Proteins
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Ferredoxins
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Iron-Sulfur Proteins