ACETYLCHOLINESTERASE FROM BOVINE ERYTHROCYTES - PURIFICATION AND PROPERTIES OF THE ENZYME SOLUBILIZED IN THE PRESENCE AND THE ABSENCE OF TRITON-X-100

  • 1 January 1979
    • journal article
    • research article
    • Vol. 34  (9-10) , 721-725
Abstract
Acetylcholinesterase was released from bovine erythrocytes by Triton X-100 treatment and purified by 2-fold affinity chromatography. The detergent-free enzyme was obtained with a specific activity of 4130 U[units]/mg (303,000-fold purification) and a 25% yield. Alternatively, the commercial available crude enzyme was purified. The latter preparation has an uniform MW 175,000. The Triton-solubilized enzyme could be resolved after removal of the detergent in 8 multiple forms (MW 175,000 and multiple values); in the presence of Triton there exists only one form (MW 338,000). Amino acid composition of the 2 enzyme preparations differed significantly. No differences were observed with respect to other properties: SDS [sodium dodecyl sulfate] gel electrophoresis revealed 2 protein bands (MW 166,000 and 86,000) with both preparations. The enzyme was a glycoprotein with a pI [isoelectric point] value of 4.3 and contains strongly bound phosphatidylethanolamine. The N-terminal amino acid was Glu (or Gln).