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Updated: Jul 11, 2026

Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry
Published on: March 24, 2012
Expression, purification, and characterization of a [Fe2S2] cluster containing ferredoxin from Acidithiobacillus
Jia Zeng1, Xia Huang, Yuandong Liu
1Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, Changsha 410083, People's Republic of China. zengjcsu@yahoo.com.cn
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.

