Differences in subunit activities in acetylcholinesterase as possible cause for apparent deviation from normal Michaelis-Menten kinetics.

1. Form Gp of acetylcholinesterase (EC 3.1.1.7) from electric eel gave curved Lineweaver-Burk plots with acetylcholine. The Hill coefficients were 0.50-0.55 at low ionic strength and increased to 0.93 with increasing ionic strength. In presence of atropine or hexamethonium normal Michaelis-Menten kinetics were observed. 2. Inhibition of the enzyme by iPr2P-F was biphasic. One half of the total enzyme activity decreased at a faster rate than the other half. With [3H]iPr2P-F, the extent of labelling was determined for the light and heavy subunits. At low [3H]iPr2P-F concentration only the light subunit was phosphorylated. Higher [3H]iPr2P-F concentrations and prolonged treatment increased the amount of label in the heavy subunit. 3. From these data it is concluded that the two subunits are labelled at different rates indicating different reactivity towards iPr2P-F as well as towards acetylcholine. These data might account for the apparent non-Michaelis-Menten type kinetics obtained at low ionic strength.