Analysis of the active center of hydrogenase from Desulfovibrio vulgaris Miyazaki by magnetic measurements.
Hydrogenase [hydrogen: ferricytochrome c3 oxidoreductase, EC 1.12.2.1] solubilized and purified from the particulate fraction of Desulfovibrio vulgaris Miyazaki F (IAM 12604) contains 8 iron and 8 labile sulfide ions in one molecule which is composed of two unequal subunits (Mr: 60,000 + 29,000). It does not contain nickel atoms. The EPR (electron paramagnetic resonance) spectrum has an isotropic signal at g = 2.017 which is independent of the temperature. The peak-to-peak width of the signal is about 20 G. The signal intensity is nearly equivalent to 1 unpaired electron per molecule. No other signals can be detected in the field range between 2,240 and 4,240 G (which corresponds to g-values between 2.91 and 1.54). Ferricyanide has only a little effect on the shape and intensity of the EPR signal. The hydrogenase reduced under H2 is EPR silent. The Mössbauer spectrum has no hyperfine splitting at 4K. The isomer shift and quadrupole splitting at 77K are 0.38 and 0.87 mm/s, respectively. Based on these magnetic measurements, the structure of the active center of hydrogenase was suggested to be [4Fe-4S]3+ + [4Fe-4S]2+.
Yagi
T
T
Kimura
K
K
Inokuchi
H
H
D003901:Desulfovibrio / Q000201:enzymology*; D004578:Electron Spin Resonance Spectroscopy; D005292:Ferricyanides / Q000494:pharmacology; D006864:Hydrogenase / Q000032:analysis*; D007501:Iron / Q000032:analysis; D008872:Microwaves; D009532:Nickel / Q000032:analysis; D013051:Spectrometry, Gamma; D013696:Temperature; D015032:Zinc / Q000032:analysis
D005292:Ferricyanides; C007931:hexacyanoferrate III; D009532:Nickel; D007501:Iron; D006864:Hydrogenase; D015032:Zinc