Mechanisms of ALA-D inhibition by lead and of its restoration by zinc and dithiothreitol.

To induce the inhibition of ALA-D (delta-aminolaevulinic acid dehydratase) activity by lead in vitro, it is necessary to preincubate the enzyme fraction with lead ions and the Hb fraction (factors) together. The combination of two of the three (ALA-D fraction, lead acetate, and Hb fraction) in the preincubation has only a small effect on the activity. Lead preincubated with ALA-D and Hb fractions does not alter the affinity of the enzyme for the substrate, suggesting that the substrate can bind to the enzyme molecules (non-competitive inhibition). The restoration of activity by zinc with dithiothreitol is associated with the removal of lead from ALA-D fraction proteins to which it has bound in vivo and in vitro. The mode of the inhibitory action of tin on ALA-D is similar to that of lead because the inhibition is intensified by the addition of Hb fraction and is restored by heating. The inhibition concentration is, however, higher than that of lead. Of the three methods recovering decreased activity, heating is the most specific to detect inhibition by lead.