Redox regulation of maize NADP-malic enzyme by thiol-disulfide interchange: effect of reduced thioredoxin on activity.
Incubation of C4 NADP-malic enzyme from maize leaves with the oxidant o-iodosobenzoate leads to the reversible and complete inactivation of the enzyme. The time-course of inactivation is biphasic with the rate depending on the o-iodosobenzoate concentration. The inactivation is partially prevented by L-malate, NADP and Mg2+ alone, while NADP plus Mg2+ afford total protection. The complete reversal of the inactivation by the reductive agents dithiothreitol and 2-mercaptoethanol suggests that the modification of the enzyme by o-iodosobenzoate occurs concomitant with the oxidation of one or more pairs of sulfhydryl groups to the disulfide state, producing a conformationally altered form of the protein or directly modifying the active site. Titration of free thiol groups before and after inactivation of maize malic enzyme by o-iodosobenzoate shows a decrease in the accessible groups from 7 to 5, suggesting inactivation is accompanied by oxidation of two vicinal thiols. The oxidized form of the enzyme is rapidly reactivated by incubation with chemical and photochemically reduced thioredoxin in vitro, while the 'dark' activity of the enzyme is enhanced to the level of the 'light' activity by dithiothreitol. This evidence suggests that a reversible reduction and oxidation of disulfide bonds may take place during the regulation of the enzyme, indicating that the redox state of the disulfide bonds of C4 NADP-malic enzyme from maize leaves is important for the expression of maximal catalytic activity.
Drincovich
M F
MF
Centro de Estudios Fotosintéticos y Bioquímicos, CONICET F.M. Lillo Universidad Nacional de Rosario, Argentina.
Andreo
C S
CS
D004220:Disulfides / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D007463:Iodobenzoates / Q000494:pharmacology*; D008291:Malate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010084:Oxidation-Reduction; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013879:Thioredoxins / Q000494:pharmacology*; D003313:Zea mays / Q000201:enzymology*
D004220:Disulfides; D007463:Iodobenzoates; D013438:Sulfhydryl Compounds; C008154:2-iodosobenzoic acid; D013879:Thioredoxins; D008291:Malate Dehydrogenase; C031917:malate dehydrogenase (oxaloacetate-decarboxylating) (NADP+)