Oxidation-reduction titration of cytochrome components in the electron transport chaim of Azotobacter vinelandii.
The multiple cytochrome components in the electron transport particle of Azotobacter vinelandii were resolved and their oxidation-reduction midpoint potentials were determined by a simultaneous potentiometric and absorption measurements under anaerobic condition with or without CO. The midpoints of the individual cytochrome component corresponding to the membrane-bound types were also determined in the solubilized fractions prepared by a differential detergent solubilization of the membrane particles of A vinelandii. Two cytochromes of b type, one with an absorption maximum measured at 559 nm and another at 561 nm in the membrane particle, were resolved and their Em, 7.4 values determined to be - 30 mV and +122mV, respectively. Cytochrome b 559 reacted in both membrane-bound and solubilized forms, however, cytochrome b561 was inert to CO treatment. Only one cytochrome of c type (c4) measured at 575-551 nm was resolved, its midpoint potential at pH 7.4 was +322mV in the membrane-bound form and +278mV in the solubilized form. This c-type cytochrome had no CO reactivity. Cytochrome d, a CO-reactive component, had a midpoint of +270mV in the membrane fraction. The midpoint of cytochrome a1 in its membrane-bound form could not be measured accurately because of its low concentration. However, in the solubilized preparations, cytochrome a1 apparently had a red shift with an absorption maximum at 613 nm, with an estimated Em, 7.4 of - 45 mV, while cytochrome d was no longer detected, possibly because of denaturation.
D001395:Azotobacter / Q000378:metabolism*; D003573:Cytochrome b Group; D003574:Cytochrome c Group / Q000378:metabolism; D017937:Cytochrome d Group; D003580:Cytochromes / Q000378:metabolism*; D045263:Cytochromes a1; D004579:Electron Transport; D010084:Oxidation-Reduction
D003573:Cytochrome b Group; D003574:Cytochrome c Group; D003580:Cytochromes; D045263:Cytochromes a1; D017937:Cytochrome d Group