title: Comparison of the "Rieske" [2Fe-2S] center in the bc1 complex and in bacterial dioxygenases by circular dichroism spectroscopy and cyclic voltammetry.
abstract: Two different types of "Rieske" [2Fe-2S] clusters have been observed in proteins, one in the bc complexes of the respiratory chain and the other in bacterial dioxygenases. We have compared the circular dichroic (CD) spectra and redox properties of the water soluble fragment of the Rieske center of the bovine heart mitochondrial bc1 complex (ISF) and of the ferredoxin from benzene dioxygenase in Pseudomonas putida ML2 (FDBED). Spinach ferredoxin was also measured for comparison. The redox potential of both proteins could be determined in solution by cyclic voltammetry (CV) and by CD-monitored spectroelectrochemistry using a specially constructed optically transparent thin layer (OTTLE) cell. Whereas the redox potential of the ISF (+312 +/- 5 mV at pH 7.0) depended both on the pH above pH 7 and on the ionic strength, the redox potential of the FDBED (-155 +/- 5 mV at pH 7.0) was observed to be independent of pH and ionic strength. The ISF showed a marked dependence of its redox potential on temperature, while the FDBED showed no temperature dependence. The entropy of the redox reaction delta S degrees rc was calculated as -88 +/- 11 J K-1 mol-1 for the bc1 Rieske center and approximately 0 J K-1 mol-1 for the FdBED. The CD spectra of Rieske type clusters are significantly different from those of plant type [2Fe-2S] ferredoxins. A strong negative CD band is present at 20 000 cm-1 (500 nm) in all reduced Rieske clusters. The possible assignment of this band is discussed as arising from the highest energy magnetically allowed d --> d transition (dz2 --> dxz) of the FeII site. If so, this band is highly indicative of the distortion of the ligand field of the FeII site.
authors.affiliation: Universitätsklinikum Frankfurt, Frankfurt/Main, Germany. Link@zbc.klinik.unifrankfurt.de
authors.forename: T A
authors.initials: TA
authors.lastname: Link
authors.forename: O M
authors.initials: OM
authors.lastname: Hatzfeld
authors.forename: P
authors.initials: P
authors.lastname: Unalkat
authors.forename: J K
authors.initials: JK
authors.lastname: Shergill
authors.forename: R
authors.initials: R
authors.lastname: Cammack
authors.forename: J R
authors.initials: JR
authors.lastname: Mason
mesh_terms: D002151:Calorimetry; D002942:Circular Dichroism; D003001:Cloning, Molecular; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods; D004578:Electron Spin Resonance Spectroscopy; D014450:Electron Transport Complex III / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008433:Mathematics; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000378:metabolism; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D011487:Protein Conformation; D016958:Pseudomonas putida / Q000201:enzymology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D007506:Iron-Sulfur Proteins; D011994:Recombinant Proteins; C037989:Rieske iron-sulfur protein; D006899:Mixed Function Oxygenases; C019765:benzene 1,2-dioxygenase; D014450:Electron Transport Complex III