Kinetics of Sheep‐Liver Cytoplasmic Aldehyde Dehydrogenase

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Abstract
Sheep liver cytoplasmic aldehyde dehydrogenase showed little pH dependence on V [velocity] or kcat [the catalytic constant]. Some buffer anion effects were noted. The oxidation of aldehydes at pH 7.6 was quantitative but irreversible. The initial velocity data indicated a sequential mechanis for the addition of substrates. Inhibition by NADH and the product analogue 2-bromo-20phenylacetic acid, together with the known tight binding of NADH to the free enzyme, indicated an ordered mechanism with NAD+ as leading substrate. Values for the data of binding and dissociation of NAD+ were obtained from the steady-state data. The values obtained were virtually identical with those which could be calculated from the data for the horse liver cytoplasmic enzyme. Close similarities were apparent for the horse and sheep liver cytoplasmic enzymes and with other tissue aldehyde dehydrogenases. Apparent substrate activation was observed with high concentrations of both acetaldehyde and propionaldehyde, a limiting value of 0.25 s-1 being obtained for Kcat. No isotope effect was observed on using [1-2H]propionaldehyde as substrate suggesting that NADH release might be rate-limiting in the steady-state. The implications of the non-linear steady-state behavior were discussed.

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