Inhibition of erythrocyte membrane (Ca2+ + Mg2+)-ATPase by the organophosphorus insecticides parathion and methylparathion.

Organophosphorus insecticides parathion and methylparathion non-competitively inhibited the activity of (Ca2+ + Mg2+)-ATPase bound to and solubilized from pig erythrocyte membrane. Both enzyme preparations exhibited biphasic substrate curves displaying the existence of two functional active sites with low and high affinity to ATP. Also, the relationship between the activity of bound enzyme and Ca2+ concentration was biphasic. The activity reached maximum at 20 microM then dropped progressively as the Ca2+ concentration was raised. The inhibition of the activity was more pronounced for parathion than for methylparathion and the solubilized enzyme preparation was more affected than the bound one. The inhibition constants (Ki) for parathion for bound enzyme were 55 and 158 microM for high- and low-affinity active sites, respectively; for methylparathion these values equalled 74 and 263 microM, respectively. Ki values for parathion were 36 and 118 microM for solubilized enzyme (high- and low-affinity sites, respectively), for methylparathion -62 and 166 microM, respectively. The magnitude of the effect was greater for a low Ca2+ concentration, which could arise from different conformational states of the enzyme at different calcium concentrations. The results of the experiment suggest that the insecticides inhibited the ATPase by binding to a site on the enzyme rather than by the interaction with associated lipids, although lipids could weaken the action of the compounds due to the strong affinity of organophosphorus insecticides to lipids.