Brain Acetylcholinesterase: Solubilization and Partial Purification by Affinity Chromatography.
Technical rept. Mar-Sep 71,
EDGEWOOD ARSENAL MD
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Guinea pig brain acetylcholinesterase was solubilized by addition of Triton X-100. Affinity chromatography was used to partially purify the solubilized enzyme. Two specific competitive inhibitors, para-carboxyphenyltrimethyl ammonium iodide and meta-carboxyphenyltrimethyl ammonium iodide, were synthesized. Sodium chloride, choline chloride, physostigmine, and pyridinium 2-aldoxime methiodide were used as eluants. Of the inhibitors synthesized, the meta-carboxyphenyltrimethyl ammonium iodide analog gave the better results. A sodium chloride elution followed by a linear gradient of choline chloride provided the best elution of the bound enzyme. The partially purified enzyme hydrolyzed 0.01 moles of 14C-acetylcholine per hour per milligram protein, a 1000-fold purification. Author