The influence of ionic strength on the interaction of quaternary drugs with mammalian acetylcholinesterase in relation to possible regulatory effects.

Abstract

Abstract— The effects of inorganic salts, gallamine triethiodide and (+)‐tubocurarine chloride on mammalian acetylcholinesterase (AChE) were examined. The results were obtained mainly from soluble erythrocyte AChE; particle‐bound and detergent‐solubilized rat brain enzymes were also used. Three aspects of AChE were examined, namely direct effects on activity, the recovery of activity of the diethylphosphorylated enzyme and thirdly the aggregation of the enzyme at low ionic strength as shown by chromatography on columns of Sepharose 6B. The action of gallamine on AChE was controlled by the substrate concentration and the ionic strength of the medium. Both inhibition and activation by gallamine could be observed, depending on the particular conditions used. All effects of gallamine disappeared when the ionic strength was raised to 015. The action of gallamine closely resembled the result of increasing ionic strength by adding NaCl, for in both cases the apparent affinity of AChE for substrate decreased and concomitantly the maximum velocity of hydrolysis increased. The phosphorylated enzyme recovered activity more rapidly when gallamine or tubocurarine were present, or when the ionic strength was increased. Aggregation of all enzyme forms was observed at low ionic strength; an increase to I= 015 dissociated AChE to the single molecular form. It was concluded that the mammalian enzymes closely resembled electric eel AChE. The possible methods by which regulation of AChE activity could occur are discussed in relation to these results.