Journal of Solid Phase Biochemistry

, Volume 3, Issue 2, pp 107–117 | Cite as

Activation of prephenate dehydratase by adsorption to agarose derivatives

  • Patricia Campbell
  • George I. Glover


A series of potential affinity adsorbents for prephenate dehydratase were synthesized by coupling allosteric effectors to agarose with and without spacer arms. Only cationic ligands containing a hexamethylene spacer arm adsorbed the enzyme. Activity could be removed from aminohexyl- and glycyl-aminohexyl-agaroses with 0.5 M NaCl (with loss of activity), 0.4 M phosphate, or 140 mM hexanediamine, but was not removed from ala-aminohexyl-, phe-aminohexyl-, met-aminohexyl-, leu-aminohexyl-, or phenylpyruvyl-aminohexyl-agaroses or by Gly, Ala, Phe (inhibitor), Met or Leu (both activators). Urea (1.0 M), 1 mM phosphate, 50% glycerol in phosphate or prephenic acid (substrate) did not release enzymatic activity from any of the gels. The gel-bound enzyme was found to have enhanced activity comparable to that of the methionine-activated soluble enzyme. The site and mode of the enzyme immobilization are discussed in relation to the possible hydrophobic/ionic subunit-subunit interactions.


Hydrophobic Amino Acid Standard Buffer Phenylpyruvate Aminohexyl Hexanediamine 
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  1. 1.
    Jensen, R. A. (1969) J. Biol. Chem. 244: 2816–2823.Google Scholar
  2. 2.
    Nester, E. W., andJensen, R. A. (1966) J. Bacteriol. 91: 1594–1598.Google Scholar
  3. 3.
    Rebello, J. L., andJensen, R. A. (1970) J. Biol. Chem. 245: 3738–3744.Google Scholar
  4. 4.
    Kane, J. F., Stenmark, S. L., Calhoun, D. H., andJensen, R. A. (1971) J. Biol. Chem. 246: 4308–4318.Google Scholar
  5. 5.
    Pierson, D. L., andJensen, R. A. (1974) J. Mol. Biol. 90: 563–579.CrossRefGoogle Scholar
  6. 6.
    Cotton, R. G. H., andGibson, F. (1965) Biochim. Biophys. Acta 100: 76–88.Google Scholar
  7. 7.
    Tiner-Harding, T., Glover, G. I., andCampbell, P. (1979) Prep. Biochem. 9: 33–41.CrossRefGoogle Scholar
  8. 8.
    Jensen, R. A. (1968) Genetics 60: 707–717.Google Scholar
  9. 9.
    Nishikawa, A. H., andBailon, P. (1976) J. Solid-Phase Biochem. 1: 33–49.CrossRefGoogle Scholar
  10. 10.
    Cautrecasas, P. (1970) J. Biol. Chem. 245: 3059–3065.Google Scholar
  11. 11.
    Zervas, L., Borovas, D., andGasiz, E. (1963) J. Am. Chem. Soc. 85: 3660–3666.CrossRefGoogle Scholar
  12. 12.
    Robert-Gero, M., andWaller, P.-P. (1972) Eur. J. Biochem. 31: 315–319.CrossRefGoogle Scholar
  13. 13.
    Cautrecasas, P., andParikh, I. (1972) Biochemistry 11: 2291–2299.CrossRefGoogle Scholar
  14. 14.
    Inman, J. K., andDintzis, H. M. (1969) Biochemistry 8: 4074–4082.CrossRefGoogle Scholar
  15. 15.
    Riepl, R. G., andGlover, G. I. (1978) Arch. Biochem. Biophys. 191: 192–197.CrossRefGoogle Scholar
  16. 16.
    Campbell, P., unpublished observations.Google Scholar
  17. 17.
    O’, P., Barry, S., andGriffin, T. (1974) FEBS Lett. 43: 169–175.CrossRefGoogle Scholar
  18. 18.
    Hofstee, B. H. J. (1976) J. Macromol. Sci. Chem. A10 (1 and 2): 111–147.CrossRefGoogle Scholar
  19. 19.
    Hofstee, B. H. J. (1976)In Methods of Protein Separation, Vol. 2,Catsimpoolas, N. (ed.), Plenum Press, New York, 245–278.Google Scholar
  20. 20.
    Yon, R. J. (1972) Biochem. J. 126: 765–767.Google Scholar
  21. 21.
    Nishikawa, A. H., Bailon, P., andRamel, A. H. (1976) J. Macromol. Sci.—Chem., A10 (1+2): 149–190.CrossRefGoogle Scholar
  22. 22.
    Nishikawa, A. H., andBailon, P. (1975) Arch. Biochem. Biophys. 168: 576–584.CrossRefGoogle Scholar
  23. 23.
    Porath, J. (1976)In Analysis and Control of Immobilized Enzyme Systems,Thomas, D., andKernevez, J.-P. (eds.), American Elsevier, New York, pp. 71–80.Google Scholar
  24. 24.
    Davey, M. W., Huang, J. W., Sulkowski, E., andCarter, W. A. (1975) J. Biol. Chem. 250: 348–349.Google Scholar
  25. 25.
    Jennissen, H. P., andHeilmeyer, L. M. G. (1975) Biochemistry 14: 754–760.CrossRefGoogle Scholar
  26. 26.
    Tanford, C. (1973) The Hydrophobic Effect, Wiley, New York, pp. 126–142.Google Scholar
  27. 27.
    Butler, L. G. (1975) Arch. Biochem. Biophys. 171: 645–650.CrossRefGoogle Scholar
  28. 28.
    Chibata, I., Tosa, T., Sato, T., andMori, T. (1976)In Methods in Enzymology, Vol. 44,Mosbach, K. (ed.), Academic Press, New York, pp. 746–759.Google Scholar
  29. 29.
    Marshall, D. L. (1973)In Immobilized Biochemicals and Affinity Chromatography,Dunlap, R. B. (ed.), Plenum Press, New York, pp. 345–368.Google Scholar

Copyright information

© Humana Press Inc. 1979

Authors and Affiliations

  • Patricia Campbell
    • 1
  • George I. Glover
    • 1
  1. 1.Department of ChemistryTexas A&M UniversityTexas

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