Neurochemical Research

, Volume 5, Issue 9, pp 943–962 | Cite as

Purification and immunochemical properties of choline acetyltransferase from human brain

  • J. H. Peng
  • P. L. McGeer
  • H. Kimura
  • S. C. Sung
  • E. G. McGeer
Original Articles

Abstract

Choline acetyltransferase (CAT) was purified to homogeneity from 363 g of human neostriatum by means of ammonium sulfate and protamine sulfate fractionation, followed by chromatography on DEAE-Sephadex, hydroxyapatite, phosphocellulose, and agarose-hexane-Co A columns. The final product migrated as a single component on 7.5% gels with or without SDS. It had a molecular weight of 66,000 daltons and a specific activity of 7.3 μmol acetylcholine formed per milligram protein per minute. Antibodies prepared in rabbits gave single precipitin lines against this protein on Ouchterlony immunodiffusion and immunoelectrophoresis plates. The CAT-anti-CAT IgG complex migrated as a single band on gel electrophoresis, establishing the monospecificity of the antibodies. Strong cross-reactivity to the IgG was obtained with CAT from rat, rabbit, and guinea pig, but only weak reactivity with chicken. Fab fragments were prepared from the rabbit IgG and were used to stain CAT-containing neurons in the spinal cord and nerve endings at the neuromuscular junction using the PAP technique.

Keywords

Acetylcholine Hydroxyapatite Ammonium Sulfate Nerve Ending Protamine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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Copyright information

© Plenum Publishing Corporation 1980

Authors and Affiliations

  • J. H. Peng
    • 1
  • P. L. McGeer
    • 1
  • H. Kimura
    • 1
  • S. C. Sung
    • 1
  • E. G. McGeer
    • 1
  1. 1.Kinsmen Laboratory of Neurological Research Department of PsychiatryUniversity of British ColumbiaBritish ColumbiaVancouverCanada

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