Affinity chromatography of acetylcholinesterase. The use of Amberlite CG-120 for dissociating the enzyme-inhibitor complex.
Acetylcholinesterase from rat brain was solubilized with 1% (w/v) Triton X-100 and purified by affinity chromatography. Two different ligands were investigated. The most efficient purification was obtained when the enzyme was eluted from a column containing the acetylcholinesterase inhibitor N-methyl-3-aminopyridinium iodide covalently linked to Sepharose 2B. An initial recovery of 6% of the applied enzyme increased to 70% after treatment with Amberlite CG-120. The partially purified enzyme had a specific activity of 205 mumoles min-1 mg-1 and a purification of 162-fold with respect to the brain homogenate and 44-fold with respect to the Triton solubilized enzyme. The effect of metal cations on the stability of the partially purified enzyme during storage at --20 degrees C was also investigated. The addition of MgCl2 to the purified enzyme prevented the rapid loss of enzyme activity.
D000110:Acetylcholinesterase / Q000302:isolation & purification*; D000818:Animals; D001921:Brain / Q000201:enzymology; D002411:Cation Exchange Resins; D002800:Cholinesterase Inhibitors; D002846:Chromatography, Affinity / Q000379:methods; D009419:Nerve Tissue Proteins / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012117:Resins, Synthetic
D002411:Cation Exchange Resins; D002800:Cholinesterase Inhibitors; D009419:Nerve Tissue Proteins; D012117:Resins, Synthetic; C008909:Amberlite CG-120; D000110:Acetylcholinesterase