Skip to main content
SpringerLink
Log in

Studies on detergent released choline acetyltransferase from membrane fractions of rat and human brain

  • Original Articles
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

The relationship between soluble and membrane choline acetyltransferase (ChAT) was studied. Differential solubilization of rat and human brain yielded ChAT in the soluble and membrane fractions. The addition of 1% Triton X-100 to membrane fractions resulted in a release of ChAT. A comparable release of lactate dehydrogenase was also observed. The Triton released ChAT and soluble ChAT from rat and human brain were efficiently purified by immuno-affinity chromatography. A single molecular weight of 68,000 was observed for both forms of rat and human brain ChAT. Epitope maps produced from both forms of human brain ChAT were identical. It is concluded that Triton release ChAT is identical to soluble ChAT and simply represents occluded soluble ChAT.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Davies, P. 1979. Neurotransmitter-related enzymes in senile dementia of the Alzheimer's type. Brain Res. 171:319–327.

    Google Scholar 

  2. Fonnum, F. 1966. Is choline acetyltransferase present in synaptic vesicles? Biochem. Pharmacol. 15:1641–1643.

    Google Scholar 

  3. Fonnum, F. 1968. Choline acetyltransferase binding to and release from membranes. Biochem. J. 109:389–397.

    Google Scholar 

  4. Whittaker, V. P., Michaelson, I. A., and Kirkland, R. J. A. 1964. The separation of synaptic vesicles from nerve-ending particles (‘synaptosomes’). Biochem. J. 90:293–303.

    Google Scholar 

  5. Tucek, S. 1966. On subcellular localization and binding of choline acetyltransferase in the cholinergic nerve endings of the brain. J. Neurochem. 13:1317–1327.

    Google Scholar 

  6. Fonnum, F. 1967. The compartmentation of choline acetyltransferase within the synaptosome. Biochem. J. 103:262–270.

    Google Scholar 

  7. Benishin, C. G., and Carroll, P. T. 1983. Multiple forms of choline-o-acetyltransferase in mouse and rat brain: solubilization and characterization. J. Neurochem. 41:1030–1039.

    Google Scholar 

  8. Eder-Colli, L., and Amato, S. 1985. Membrane-bound choline acetyltransferase in torpedo electric organ; a marker for synaptosomal plasma membranes? Neuroscience, 15:577–589.

    Google Scholar 

  9. Peng, J. H., McGeer, E. G., and McGeer, P. L. 1984. Purification of membrane bound choline acetyltransferase from human brain. In Proceedings of the 14th Annual Meeting of the Society for Neuroscience, Anaheim, California, October, 1984. Abstract 342.5.

  10. Peng, J. H., McGeer, P. L., and McGeer, E. G. 1986. Membrane-bound choline acetyltransferase from human brain: purification and properties. Neurochem. Res. 11:959–971.

    Google Scholar 

  11. Badamchian, M., and Carroll, P. T. 1985. Molecular weight determinations of soluble and membrane-bound fractions of choline-o-acetyltransferase in rat brain. J. Neurosci. 5:1955–1964.

    Google Scholar 

  12. Tuček, S. 1984. Problems in the organization and control of acetylcholine synthesis in brain neurons. Prog. Biophys. Molec. Biol. 44:1–46.

    Google Scholar 

  13. Peng, J. H., McGeer, P. L., and McGeer, E. G. 1983. Anti-human choline acetyltransferase fragments antigen binding (Fab)-sepharose chromatography for enzyme purification. Neurochem. Res. 8:1481–1486.

    Google Scholar 

  14. McGeer, P. L., McGeer, E. G., and Peng, J. H. 1984. Choline acetyltransferase: purification and immunohistochemical localization. Life Sciences 34:2319–2338.

    Google Scholar 

  15. Bruce, G., Wainer, B. H., and Hersh, L. B. 1985. Immunoaffinity purification of human choline acetyltransferase: Comparison of the brain and placental enzymes. J. Neurochem. 45:611–620.

    Google Scholar 

  16. Fonnum, F. 1975. A rapid radiochemical method for the detection of choline acetyltransferase. J. Neurochem. 24:407–409.

    Google Scholar 

  17. Johnson, M. K. 1960. The intracellular distribution of glycolytic and other enzymes in rat brain homogenates and mitochondrial preparations. Biochem. J. 77:610–618.

    Google Scholar 

  18. Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 71:248–254.

    Google Scholar 

  19. Levey, A. I., Aoki, M., Fitch, F. W., and Wainer, B. H., 1981. The production of monoclonal antibodies reactive with bovine choline acetyltransferase. Brain Res. 218:383–387.

    Google Scholar 

  20. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685.

    Google Scholar 

  21. Hersh, L. B., Wainer, B. H., and Andrews, L. 1984. Multiple isoelectric and molecular weight variants of choline acetyltransferase: artifact or real? J. Biol. Chem. 259:1253–1258.

    Google Scholar 

  22. Cleveland, D. W., Fischer, S. G., Kirschner, M. W., and Laemmli, V. K. 1977. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J. Biol. Chem. 252:1102–1106.

    Google Scholar 

  23. Beattie, D. S., Sloan, H. R., and Basford, R. E. 1963. Brain mitochondria II. The relationship of brain mitochondria to glycolysis. J. Cell Biol. 19:309–316.

    Google Scholar 

Download references

Author information

Author notes

  1. Gordon Bruce

    Present address: Howard Hughes Medical Institute, Massachusetts General Hospital, Wellman 3, 02114, Boston, MA

Authors and Affiliations

  1. Department of Biochemistry, University of Texas Health Science Center at Dallas, 5323 Harry Hines Blvd., 75235, Dallas, Texas

    Gordon Bruce & Louis B. Hersh

Authors
  1. Gordon Bruce
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Louis B. Hersh
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bruce, G., Hersh, L.B. Studies on detergent released choline acetyltransferase from membrane fractions of rat and human brain. Neurochem Res 12, 1059–1066 (1987). https://doi.org/10.1007/BF00971705

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00971705

Key Words

Search

Navigation