Brain Acetylcholinesterase: Solubilization and Partial Purification by Affinity Chromatography.
Abstract
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)
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 1972
- Accession Number
- AD0750111
Entities
People
- Clarence A. Broomfield
- Douglas W. Reichard
- Henry I. Yamamura
- Tommy L. Gardner