Purification and Characterization of the 2-Oxoglutarate Dehydrogenase Complex from Dictyostelium discoideum

The 2-oxoglutarate dehydrogenase complex was isolated from the cellular slime mould, Dictyostelium discoideum, and purified 113-fold. The enzyme exhibited Michaelis-Menten kinetics and the K m values for 2-oxoglutarate, CoA, and NAD were 1·0 mM, 0·002 mM, and 0·07 mM, respectively. The K i value for succinyl-CoA was determined to be 0·004 mM and the K i for NADH was 0·018 mM. AMP had positive effects whereas ATP had negative effects on the enzyme activity. The kinetic constants determined in this study and the reaction mechanism suggested can now be incorporated into a transition model of the tricarboxylic acid cycle during differentiation of D. discoideum.