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A rapid method for the purification of octopine dehydrogenase for the determination of cell metabolites

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Summary

Unlike other NAD+-dependent dehydrogenases, octopine dehydrogenase was not bound by blue Sepharose. A rapid 2-step purification procedure (gel filtration on Sephadex G-100 followed by affinity chromatography on blue Sepharose) resulted in a final preparation of octopine dehydrogenase which had a sp. act. of 65 units/mg protein and was free of contaminating NAD+-oxidoreductases. This preparation has been used successfully for the estimation of phospho-L-arginine, L-arginine and octopine in perchloric acid extracts.

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References

  1. G. Gäde and M. Grieshaber, Analyt. Biochem.66, 393 (1975).

    Article  Google Scholar 

  2. M. Grieshaber, Analyt. Biochem.74, 600 (1976).

    Article  CAS  PubMed  Google Scholar 

  3. M. Grieshaber, E. Kronig and R. Koorman, Hoppe-Seylers Z. physiol. Chem.359, 133 (1978).

    CAS  PubMed  Google Scholar 

  4. A. Olomucki, C. Huc, F. Lefebure and N.V. Thoai, Eur. J. Biochem.28, 261 (1972).

    Article  CAS  PubMed  Google Scholar 

  5. G. Gäde and M. Grieshaber, J. comp. Physiol.102, 149 (1975).

    Article  Google Scholar 

  6. G. Gäde and M. Grieshaber, Archs int. Physiol. Biochim.84, 735 (1976).

    Google Scholar 

  7. N.V. Thoai, C. Huc, D.B. Pho and A. Olomucki, Biochim. biophys. Acta191, 46 (1969).

    Article  PubMed  Google Scholar 

  8. Blue Sepharose CL-6B is commercially available from Pharmacia Fine Chemicals AB.

  9. S.T. Thompson, K.H. Cass and E. Stellwagen, Proc. natl. Sci. USA72, 669 (1975).

    Article  CAS  Google Scholar 

  10. R.L. Easterday and I. Easterday, in: Immobilized Biochemicals and Affinity Chromatography, p. 123. Ed. R.B. Dunlap. Plenum Press, New York 1974.

    Chapter  Google Scholar 

  11. H.-J. Böhme, G. Kopperschläger, J. Schulz and E. Hofman, J. Chromatogr.69, 209 (1972).

    Article  PubMed  Google Scholar 

  12. A. Szent-Györgyi, C. Cohen and J. Kendrick-Jones, J. molec. Biol.56, 239 (1971).

    Article  PubMed  Google Scholar 

  13. F. Regnouf and N.V. Thoai, Comp. Biochem. Physiol.32, 411 (1970).

    Article  CAS  PubMed  Google Scholar 

  14. G. Gäde, E. Weeda and P.A. Gabbott, J. comp. Physiol.124, 121 (1978).

    Article  Google Scholar 

Download references

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Author notes

  1. G. Gäde

    Present address: Institut für Zoophysiologie, AVZ I, Rheinische Friedrich-Wilhelms-Universität, Endenicher Allee 11-13, D-5300, Bonn, Federal Republic of Germany

Authors and Affiliations

  1. N.E.R.C. Unit of Marine Invertebrate Biology, Marine Science Laboratories, Menai Bridge, Gwynedd, UK

    G. Gäde & E. J. H. Head

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  1. G. Gäde
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  2. E. J. H. Head
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Supported by a grant from the Deutsche Forschungsgemeinschaft (Ga 241/1).

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Gäde, G., Head, E.J.H. A rapid method for the purification of octopine dehydrogenase for the determination of cell metabolites. Experientia 35, 304–305 (1979). https://doi.org/10.1007/BF01964314

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