The oxygen-dependent deactivation and reactivation of spinach ribulose-1,5-bisphosphate carboxylase.
Ribulose-1,5-bisphosphate carboxylase-oxygenase (3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39) is deactivated by the removal of oxygen, and reversibly reactivated by its readdition to the enzyme solution. A short pulse of oxygen to the anaerobic enzyme solution is sufficient to trigger the reactivation process; the Ka value for this reaction was estimated as 0.12 mM oxygen. The enzyme could not be reactivated under anaerobic conditions by an organic oxidant (benzoylperoxide) or by sulfhydryl group reducing reagents (dithiothreitol or beta-mercaptoethanol), suggesting that the process of reactivation was oxygen specific. Furthermore, the inhibition of the reactivation by superoxide anion scavengers such as Tiron (1,2-dihydroxybenzene-3,5-disulfonic acid), copper penicillamine, hydroxylamine, nitroblue tetrazolium, and ascorbate, indicated that the monovalent reduced oxygen was involved as the reacting species in this process. The deactivation of the enzyme associated with the removal of oxygen was also sensitive to the presence of scavengers of O2(-), suggesting that superoxide anion was also involved in the deactivation process. Both the carboxylase and the oxygenase activities were similarly affected under all the experimental conditions studied. On the basis of these results it is argued that the enzyme molecules are able to reduce oxygen and that superoxide anion causes the deactivation or reactivation of the enzyme.
D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt / Q000494:pharmacology; D001585:Benzoyl Peroxide / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology*; D010944:Plants / Q000201:enzymology*; D012273:Ribulose-Bisphosphate Carboxylase / Q000378:metabolism*; D013481:Superoxides / Q000494:pharmacology
D013481:Superoxides; D014013:1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt; D008623:Mercaptoethanol; D002262:Carboxy-Lyases; D012273:Ribulose-Bisphosphate Carboxylase; D010100:Oxygen; D004229:Dithiothreitol; D001585:Benzoyl Peroxide