PMID:18286376
| Citation |
Boxhammer, S, Glaser, S, Kühl, A, Wagner, AK and Schmidt, CL (2008) Characterization of the recombinant Rieske [2Fe-2S] proteins HcaC and YeaW from E. coli. Biometals 21:459-67 |
|---|---|
| Abstract |
Three genes within the genome of E. coli K12 are predicted to encode proteins containing the typical Rieske iron-sulfur cluster-binding motifs. Two of these, hcaC and yeaW, were overexpressed in E. coli BL21 and Tuner (DE3) pLacI. The recombinant proteins were purified and analyzed by UV/Vis- and EPR-spectroscopy. HcaC and YeaW display the typical redox-dependent UV/Vis-spectra of iron-sulfur proteins. The EPR spectrum of reduced HcaC shows characteristic g-values of a Rieske cluster whereas the g-values for YeaW are close to the upper limit for this type of iron-sulfur cluster. Both iron-sulfur clusters could be reduced by dithionite, but not by ascorbate, confirming their classification as low-potential Rieske proteins as derived from the amino acid sequences. A phylogenetic analysis of the two proteins reveals that HcaC clearly segregates with the Rieske ferredoxins of class IIB oxygenases whereas the classification of YeaW remains doubtful. |
| Links |
PubMed Online version:10.1007/s10534-008-9134-y |
| Keywords |
Amino Acid Sequence; Escherichia coli/metabolism; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/classification; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Ferredoxins/chemistry; Ferredoxins/classification; Ferredoxins/genetics; Ferredoxins/metabolism; Iron-Sulfur Proteins/chemistry; Iron-Sulfur Proteins/classification; Iron-Sulfur Proteins/genetics; Iron-Sulfur Proteins/metabolism; Models, Molecular; Molecular Sequence Data; Oxidoreductases/chemistry; Oxidoreductases/classification; Oxidoreductases/genetics; Oxidoreductases/metabolism; Phylogeny; Protein Structure, Tertiary; Recombinant Proteins/chemistry; Recombinant Proteins/classification; Recombinant Proteins/genetics; Recombinant Proteins/metabolism; Sequence Alignment; Sequence Analysis, Protein |
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