The reduction of flavocytochrome b2 by carboxylate radicals. A pulse radiolysis study.
The reaction of flavocytochrome b2 with carboxylate radicals has been studied by the pulse-radiolysis technique. The absorbance difference spectra observed 2, 10 and 90 mus after the pulse were very similar to the (reduced-minus-oxidized) difference spectrum of the cytochrome b2 core, a deflavoderivative prepared from flavocytochrome b2. The heme b2 group was reduced in a bimolecular reaction with a rate constant of (2.1 +/- 0.2) X 10(8) M-1 X s-1. Simulation data allowed us to assign this reduction mainly to a direct reaction of CO-2 on heme b2 without flavin involvement. However, this heme b2 reduction was accelerated in a flavocytochrome b2 sample of low molar activity. This observation seemed to reflect modifications of the heme b2 environment, in particular a closer contact with the solvent. Moreover, in such samples the flavin became reducible to the semiquinone state as deduced from absorbance difference spectra in the 440-490 nm region. The agreement between experimental and computed time-courses allowed to estimate the reaction rate of bound flavin to be equal to 2 X 10(8) M-1 X s-1 in this studied sample. Thus the reactivity of bound heme b2 and flavin within flavocytochrome b2 with CO-2 seems to be sensitive to the physical alterations of the polypeptide chain.
D002264:Carboxylic Acids; D005609:Free Radicals; D007700:Kinetics; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome); D010084:Oxidation-Reduction; D010843:Pichia / Q000201:enzymology; D013057:Spectrum Analysis
D002264:Carboxylic Acids; D005609:Free Radicals; D007770:L-Lactate Dehydrogenase; D042844:L-Lactate Dehydrogenase (Cytochrome)