Acetylcholinesterase release from mammalian erythrocytes by phosphatidylinositol-specific phospholipase C of Bacillus thuringiensis and characterization of the released enzyme.
The mode of acetylcholinesterase release from mammalian erythrocyte membranes by the action of phosphatidylinositol(PI)-specific phospholipase C of Bacillus thuringiensis was studied. As regards intact erythrocytes, a larger amount of acetylcholinesterase was released from sheep or bovine erythrocytes than from horse erythrocytes. From horse erythrocyte ghosts, acetylcholinesterase was more easily released than from intact cells. Bovine erythrocyte acetylcholinesterase released by PI-specific phospholipase C was purified by column chromatography on DEAE-cellulose, affinity gel and Sepharose 6B, to a homogeneous state, as indicated by polyacrylamide gel electrophoresis, with a recovery of 39%. Also, bovine erythrocyte acetylcholinesterase was solubilized by Triton X-100 and partially purified. The properties of these acetylcholinesterase preparations obtained by the action of PI-specific phospholipase C and/or Triton X-100 were studied in detail. On elution from the Sepharose 6B column, Triton X-100-solubilized acetylcholinesterase was eluted at the void volume while the enzyme obtained by further treatment with PI-specific phospholipase C was eluted in the region corresponding to M.W. 250,000. Furthermore, the heat stability of acetylcholinesterase purified after solubilization with PI-specific phospholipase C was higher than that of the Triton X-100-solubilized acetylcholinesterase. The close association and direct interaction of PI with acetylcholinesterase in the erythrocyte membrane was suggested by the above results.
D000110:Acetylcholinesterase / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D000818:Animals; D001413:Bacillus thuringiensis / Q000201:enzymology*; D002417:Cattle; D004355:Drug Stability; D004910:Erythrocyte Membrane / Q000201:enzymology; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007700:Kinetics; D010716:Phosphatidylinositols / Q000378:metabolism; D010740:Phospholipases / Q000378:metabolism*; D012756:Sheep; D013816:Thermodynamics; D010738:Type C Phospholipases / Q000378:metabolism* / Q000494:pharmacology
D010716:Phosphatidylinositols; D010740:Phospholipases; D000110:Acetylcholinesterase; D010738:Type C Phospholipases