The activation of ribulose-1,5-bisphosphate carboxylase-oxygenase from spinach by oxygen.
The effect of oxygen on ribulose-1,5-bisphosphate carboxylase-oxygenase from spinach was investigated. Both activities were deactivated by removal of oxygen and reversibly reactivated by oxygenation of the enzyme solution. The change in enzyme activities was accompanied by conformational changes as studied by the use of intrinsic and extrinsic fluorescent probes. The analysis of cysteine sulfhydryl groups accessible to 5,5'-dithiobis-(2-nitrobenzoic acid) revealed that the number of these groups changed with the oxygen concentration. The kinetic of the exposure of eight cysteine residues was similar to the loss of enzyme activities. The modification of these groups with 5,5'-dithiobis-(2-nitrobenzoic acid) caused almost complete loss of both the activities. The enzyme isolated from a photolithotrophic organism, Chromatium vinosum, was not affected by oxygen removal. During air--argon transitions, neither the enzyme conformation nor the number of accessible sulfhydryl groups changed.