Neurochemical Research

, Volume 13, Issue 1, pp 31–35 | Cite as

Preparation and purification of γγ enolase (neuron-specific enolase) using high performance anion exchange chromatography

  • Jean-Louis Viallard
  • M. R. Ven Murthy
  • Bernard Dastugue
Original Articles


A simple and rapid method, using only two chromatographic steps, is described for the purification and preparation of γγ enolase isoenzymes from human and beef brain extracts. In the first step, a crude γγ enolase was obtained by chromatography on Q-Sepharose Fast Flow column. The crude fraction was then purified by high performance anion exchange chromatography on a Mono-Q column. γγ enolase obtained in this manner was shown to be homogeneous by two dimensional polyacrylamide gel electrophoresis and by high performance gel permeation chromatography. The yield of γγ enolase by this method was 7–8 mg of pure enzyme per 100 g of brain.

Key Words

γγ Enolase high performance anion exchange chromatography 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Mokuno, S., Kato, K., Kawai, K., Matsuoka, Y., Yanagi, T., and Sobue, I. 1983. Neuron-specific enolase and S-100 protein levels in CSF of patients with various neurological diseases. J. Neurol. Sci. 60:443–451.PubMedGoogle Scholar
  2. 2.
    Zeltzer, P. M., Marangos, P. J., Parma, A. M., Sather, H., Dalton, A., Hammond, D., Siegel, S. W., and Seeger, R. C. 1983. Raised neuron specific enolase in serum of children with metastatic neuroblastoma. Lancet 2:361–363.PubMedGoogle Scholar
  3. 3.
    Johnson, D. H., Marangos, P. J., Forbes, J. T., Hainsworth, J. D., Van Walch, R., Hande, K. R., and Greco, F. A. 1984. Potential utility of serum neuron-specific enolase in small cell carcinoma of the lung. Cancer Res. 44:5409–5414.PubMedGoogle Scholar
  4. 4.
    Grasso, A., Roda, G., Hogue-Angeletti, R. A., Moore, B. W., and Perez, V. J. 1977. Preparation and properties of the brain specific protein 14-3-2. Brain Res. 124:497–507.PubMedGoogle Scholar
  5. 5.
    Suzuki, F., Umeda, Y., and Kato, K. 1980. Rat brain enolase isoenzymes. Purification of three forms of enolase. J. Biochem. 87:1587–1594.PubMedGoogle Scholar
  6. 6.
    Hullin, D. A., Brown, K., Kynoch, A. M., Smith, C., and Thompson, R. J. 1980. Purification, radioimmunoassay and distribution of human brain 14-3-2 protein (nervous system specific enolase) in human tissues. Biochem. Biophys. Acta 628:98–108.PubMedGoogle Scholar
  7. 7.
    marangos, P. J., Zomzely-Neurath, C., Lak, D. C. M., and York, C. 1975. Isolation and characterization of the nervous system-specific protein 14-3-2 from rat brain. J. Biol. Chem. 250:1884–1891.PubMedGoogle Scholar
  8. 8.
    Keller, A., Scarna, H., Mermet, A., and Pujol, J. F. 1981. Biochemical and immunological properties of the mouse brain enolase purified by a simple method. J. Neurochem. 36:1389–1397.PubMedGoogle Scholar
  9. 9.
    Shimizu, A., Suzuki, F., and Kato, K. 1982. Purification of two enolases from human brain using a chromatofocusing column. Biochem. Biophys. Acta, 717:348–354.PubMedGoogle Scholar
  10. 10.
    Gerbitz, K. D., Denfel, T., Summer, J., Thallemer, J., and Wieland, O. H. 1983. Brain specific proteins: Creatine kinase BB isoenzymes is cochromatographed during preparation of neuron-specific enolase from human brain. Clin. Chim. Acta 133:233–239.PubMedGoogle Scholar
  11. 11.
    Rider, C. C., and Taylor, C. B. 1974. Enolase isoenzymes in rat tissues. Electrophoretic, chromatographic, immunological and kinetic properties. Biochem. Biophys. Acta 365:285–300.PubMedGoogle Scholar
  12. 12.
    Szasz, G., Gruber, W., and Bernt, E. 1976. Creatine kinase in serum: 1. Determination of optimum reaction conditions. Clin. Chem. 22:650–656.PubMedGoogle Scholar
  13. 13.
    Viallard, J. L., Murthy, M. R. V., and Dastugue, B. 1986. Rapid electrophoretic determination of neuron-specific enolase isoenzymes in serum. Clin. Chem. 32:593–597.PubMedGoogle Scholar
  14. 14.
    Somer, H., and Konttinen, A. 1972. Demonstration of serum creatine kinase isoenzymes by fluorescence technique. Clin. Chim. Acta 40:133–138.PubMedGoogle Scholar
  15. 15.
    O'Farrel, P. H. 1975. High resolution two-dimentional electrophoresis of proteins. J. Biol. Chem. 250:4007–4021.PubMedGoogle Scholar
  16. 16.
    Ishioka, M., Isobe, T., Kadoya, T., Okuyama, T., Nakajamina, T. 1984. Large scale preparation and crystallization of neuron specific enolase. J. Biochem. 95:611–617.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1987

Authors and Affiliations

  • Jean-Louis Viallard
    • 1
  • M. R. Ven Murthy
    • 2
  • Bernard Dastugue
    • 1
  1. 1.Laboratoire de Biochimie Médicale, Faculté de MédecineUniversité de Clermont-Ferrand IClermont-Ferrand CedexFrance
  2. 2.Department de Biochimie, Faculté de MédecineUniversité LavalCanada

Personalised recommendations