The inhibitory effects of polyoxyethylene detergents on human erythrocyte acetylcholinesterase and Ca2+ + Mg2+ ATPase.
The activities of acetylcholinesterase and Ca2+ + Mg2+ ATPase were measured following treatment of human erythrocyte membranes with nonsolubilizing and solubilizing concentrations of Triton X-100. A concentration of 0.1% (v/v) Triton X-100 caused a significant inhibition of both enzymes. The inhibition appears to be caused by perturbations in the membrane induced by Triton X-100 incorporation. No acetylcholinesterase activity and little Ca2+ + Mg2+ ATPase activity were detected in the supernatant at 0.05% Triton X-100 although this same detergent concentration induced changes in the turbidity of the membrane suspension. Also, no inhibition of soluble acetylcholinesterase was observed over the entire detergent concentration range. The inhibition of these enzymes at 0.1% Triton X-100 was present over an eightfold range of membrane protein in the assay indicating an independence of the protein/detergent ratio. The losses in activities of these two enzymes could be prevented by either including phosphatidylserine in the Triton X-100 suspension or using Brij 96 which has the same polyoxyethylene polar head group but an oleyl hydrophobic tail instead of the p-tert-octylphenol group of Triton X-100. The results are discussed in regard to the differential recovery of enzyme activities over the entire detergent concentration range.
Moore
R B
RB
Department of Pediatrics, University of South Alabama, Mobile 36617.
Manery
J F
JF
Still
J
J
Mankad
V N
VN
D000110:Acetylcholinesterase / Q000097:blood*; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors* / Q000097:blood; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors* / Q000097:blood; D002800:Cholinesterase Inhibitors / Q000494:pharmacology*; D003902:Detergents; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000201:enzymology; D006801:Humans; D017137:Nonoxynol; D017830:Octoxynol; D010718:Phosphatidylserines / Q000494:pharmacology; D010938:Plant Oils; D011092:Polyethylene Glycols; D012995:Solubility
D002800:Cholinesterase Inhibitors; D003902:Detergents; D010718:Phosphatidylserines; D010938:Plant Oils; D017137:Nonoxynol; D011092:Polyethylene Glycols; D017830:Octoxynol; C033084:polyethylene glycol oleyl ether; D000110:Acetylcholinesterase; D017301:Ca(2+) Mg(2+)-ATPase; D000252:Calcium-Transporting ATPases