Affinitätschromatographische Reinigung der Acetylcholinesterase aus Nucleus Caudatus vom Rind im Großmaßstab

Abstract

The acetylcholinesterase from bovine caudate nucleus was solubilized with 0.6-0.8% Triton X-100. It was purified and freed from Triton X-100 by a 2-fold affinity chromatographic procedure. A simple 3-step synthesis of the inhibitor with attached spacer arm used in affinity chromatography is described. Starting from 3 kg of caudate nucleus, nearly 11 mg of the purified acetylcholinesterase with a specific activity of 4250 U/mg (13,500-fold purification) could be obtained in an over-all yield of 49%. On gel electrophoresis the enzyme produces several enzymatically active glycoproteinbands of oligomeres. On Na dodecylsulfate gel electrophoresis in the presence of a disulfide-reducing agent the smallest subunit MW of 72,000 .+-. 2000. In the absence of a disulfide-reducing agent a band for a dimer besides the band for the monomer could be detected and a MW of 142,000 .+-. 3000 was estimated for this species.