Expression, purification and characterization of an iron-sulfur cluster assembly protein, IscU, from Acidithiobacillus ferrooxidans.
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 [Fe(2)S(2)] cluster could be assembled in apoIscU with Fe(2+) and sulfide in vitro, and in the presence of IscA and IscS, the IscU could utilize L: -cysteine and Fe(2+) to synthesize [Fe(2)S(2)] cluster in the protein. Site-directed mutagenesis for the protein revealed that Cys37, Asp39, Cys63 and Cys106 were involved in ligating with the [Fe(2)S(2)] cluster.
Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, Changsha, 410083, PR China.
Jia
J
Zeng
Wenjie
W
Zhao
Yuandong
Y
Liu
Lexian
L
Xia
Jianshe
J
Liu
Guanzhou
G
Qiu
D042763:Acidithiobacillus / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000302:isolation & purification / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010957:Plasmids; D016415:Sequence Alignment; D013056:Spectrophotometry, Ultraviolet
D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D050505:Mutant Proteins