Expression, purification and characterization of an iron-sulfur cluster assembly protein, IscU, from Acidithiobacillus ferrooxidans

被引：9

作者：

Zeng, Jia ^{[1
]}

Zhao, Wenjie ^{[1
]}

Liu, Yuandong ^{[1
]}

Xia, Lexian ^{[1
]}

Liu, Jianshe ^{[1
]}

Qiu, Guanzhou ^{[1
]}

机构：

[1] Cent S Univ, Sch Resources Proc & Bioengn, Dept Bioengn, Changsha 410083, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2007年 / 29卷 / 12期

关键词：

acidithiobacillus ferrooxidans; expression; His-tag; iron-sulfur cluster; IscU; purification;

D O I：

10.1007/s10529-007-9488-1

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 [微生物学]; 0836 [生物工程]; 090102 [作物遗传育种]; 100705 [微生物与生化药学];

摘要：

An iron-sulfur cluster assembly protein, IscU, is encoded by the operon iscSUA in Acidithiobacillus ferrooxidans. The gene of IscU was cloned and expressed in Escherichia coli. The protein was purified by one-step affinity chromatography to homogeneity. The protein was in apo-form, the [Fe2S2] cluster could be assembled in apoIscU with Fe2+ and sulfide in vitro, and in the presence of IscA and IscS, the IscU could utilize L-cysteine and Fe2+ to synthesize [Fe2S2] cluster in the protein. Site-directed mutagenesis for the protein revealed that Cys37, Asp39, Cys63 and Cys106 were involved in ligating with the [Fe2S2] cluster.

引用

页码：1965 / 1972

页数：8

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