Metal Ion-promoted Dye-ligand Chromatography

  • P. Hughes
  • R. F. Sherwood


Over the last decade, triazine dyes have received widespread attention as tools in protein purification technology (Lowe et al., 1981). It is only in more recent years, however, that the potential of multivalent metal ions in promoting protein binding to triazine dye affinity adsorbents was recognised (Hughes et al., 1982a). Previously, little reference had been made to the role of metal ions in triazine dye affinity chromatography. Only the influence of Mg2+ had received some attention, for example, as a requirement to achieve maximum binding of yeast hexokinase to immobilised Procion Green H4-G (Clonis et al., 1981). Enzyme was subsequently eluted using buffers from which the metal ions had been omitted. High concentrations of Mg2+ were also used during the purification of isoleucyl and glycyl tRNA synthetases on Blue Dextran—Sepharose (Moe and Piszkiewicz, 1976). In this case the isoleucyl enzyme was eluted with an ATP gradient after Mg2+ had been removed by washing with the equilibration buffer, while glycyl tRNA synthetase was eluted using a decreasing Mg2+ gradient. The binding of frog liver carbamyl phosphate synthetase I (Mori and Cohen, 1978) and isocitrate dehydrogenase (Nagaoka et al., 1977) to immobilised Cibacron Blue F3G-A had also been shown to be influenced by Mg2+; these enzymes were eluted with Mg2+-free buffer and the chelating agent EDTA, respectively.


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  1. Ashton, A. and Polya, G. (1978). Biochem. J., 175, 501CrossRefGoogle Scholar
  2. Baird, J. K., Sherwood, R. F., Carr, R. J. G. and Atkinson, A. (1976). FEBS Lett., 70, 61CrossRefGoogle Scholar
  3. Beissner, R.S. and Rudolph, F. B. (1978). Arch. Biochem. Biophys., 189, 76CrossRefGoogle Scholar
  4. Bollin, E. Jr. and Sulkowski, E. (1981). J. Gen. Virol., 52, 227CrossRefGoogle Scholar
  5. Bornmann, L. and Hess, B. (1977). Z. Naturforsch., 32, 756Google Scholar
  6. Borrebaeck, C. A. K., Lönnerdal, B. and Etzler, M. E. (1981). FEBS Lett., 130, 194CrossRefGoogle Scholar
  7. Bouriotis, V. and Dean, P. D. G. (1981). J. Chromatogr., 206, 521CrossRefGoogle Scholar
  8. Chen, C. C. and Somberg, E. W. (1980). Biochim. Biophys. Acta, 613, 514CrossRefGoogle Scholar
  9. Clonis, Y. D. (1982). J. Chromatogr., 236, 69CrossRefGoogle Scholar
  10. Clonis, Y. D., Goldfinch, M. J. and Lowe, C. R. (1981). Biochem. J., 197, 203CrossRefGoogle Scholar
  11. Clonis, Y. D. and Lowe, C. R. (1980). Biochem. J., 191, 247CrossRefGoogle Scholar
  12. Duckworth, H. W. and Coleman, J. E. (1970). J. Biol. Chem., 245, 1613Google Scholar
  13. Easterday, R. and Easterday, I. (1974). Adv. Exp. Med. Biol., 42, 123CrossRefGoogle Scholar
  14. Edwards, R.A. and Woody, R. W. (1979). Biochemistry, 18, 5197CrossRefGoogle Scholar
  15. Fling, M., Horowitz, N. H. and Heinemann, S.F. (1963). J. Biol. Chem., 238, 2405Google Scholar
  16. Fulton, S. (1980). In Dye Ligand Chromatography, Amicon Corp., Lexington, Mass.Google Scholar
  17. Hubert, P. and Porath, J. (1981). J. Chromatogr., 206, 164CrossRefGoogle Scholar
  18. Hughes, P., Lowe, C. R. and Sherwood, R. F. (1982a). Biochim. Biophys. Acta, 700, 90CrossRefGoogle Scholar
  19. Hughes, P., Sherwood, R. F. and Lowe, C. R. (1982b). Biochem. J., 205, 453CrossRefGoogle Scholar
  20. Hughes, P., Sherwood, R. F. and Lowe, C. R. (1984). Eur. J. Biochem., 144, 135CrossRefGoogle Scholar
  21. Irving, H. and Williams, R. J. P. (1948). Nature, Lond., 162, 746CrossRefGoogle Scholar
  22. Jacoby, W. and Wilchek, M. (1974) (Eds.). Affinity Techniques, Academic Press, New YorkGoogle Scholar
  23. Kikuchi, H. and Watanabe, M. (1981). Analyt. Biochem., 115, 109CrossRefGoogle Scholar
  24. Kitz, R. and Wilson, I. B. (1962). J. Biol. Chem., 237, 3245Google Scholar
  25. Kreuger, R. C. (1959). Fed. Proc., 18, 267Google Scholar
  26. Land, M. and Byfield, P. G. H. (1979). Int. J. Biol. Macromol., 1, 223CrossRefGoogle Scholar
  27. Lönnerdal, B. and Keen, C. L. (1982). J. Appl. Biochem., 4, 203Google Scholar
  28. Lowe, C. R., Small, D. A. P. and Atkinson, A. (1981). Int. J. Biochem., 13, 33CrossRefGoogle Scholar
  29. Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. (1951). J. Biol. Chem., 193, 265Google Scholar
  30. McArdell, J. E. C., Atkinson, T. and Bruton, C. J. (1982). Eur. J. Biochem., 125, 361CrossRefGoogle Scholar
  31. Malmström, B.G. (1957). Arch. Biochem. Biophys., 70, 58CrossRefGoogle Scholar
  32. Minton, N. P., Atkinson, T., Bruton, C. J. and Sherwood, R. F. (1984). Gene, 31, 31CrossRefGoogle Scholar
  33. Minton, N. P., Atkinson, T. and Sherwood, R. F. (1983). J. Bact., 156, 1222Google Scholar
  34. Moe, J. and Piszkiewicz, D. (1976). FEBS Lett., 72, 147CrossRefGoogle Scholar
  35. Mori, M. and Cohen, P. P. (1978). Fed. Proc., 37, 1341Google Scholar
  36. Morrill, E. M., Thompson, S. T. and Stellwagen, E. (1979). J. Biol. Chem., 254, 4371Google Scholar
  37. Nagaoka, T., Hachimora, A., Takeda, A. and Samejima, T. (1977). J. Biochem., 81, 71Google Scholar
  38. Porath, J. (1979). Coll. Inserm, 86, 17Google Scholar
  39. Porath, J., Carlsson, J., Olsson, I. and Belfrage, G. (1975). Nature, Lond., 258, 598CrossRefGoogle Scholar
  40. Porath, J. and Olin, B. (1983). Biochemistry, 22, 1621CrossRefGoogle Scholar
  41. Quinot, E. and Clays, C. (1956). C. R. Séanc. Soc. Biol., 150, 176Google Scholar
  42. Rajgopal, S. and Vijayalakshmi, M. A. (1983). J. Chromatogr., 280, 77CrossRefGoogle Scholar
  43. Rijken, D. C. and Collen, D. (1981). J. Biol. Chem., 256, 7035Google Scholar
  44. Scopes, R. K. (1982). In Affinity Chromatography and Related Techniques (ed. T. C. J. Gribnau, J. Visser and R. J. F. Nivard), pp. 333–339, Elsevier, AmsterdamGoogle Scholar
  45. Scully, M. F. and Kakker, V. V. (1982). Biochim. Biophys. Acta, 700, 130CrossRefGoogle Scholar
  46. Sherwood, R. F., Melton, R. G., Alwan, S. M. and Hughes, P. (1985). Eur. J. Biochem., 148, 447CrossRefGoogle Scholar
  47. Small, D. A. P., Atkinson, A. and Lowe, C. R. (1981). J. Chromatogr., 216, 175CrossRefGoogle Scholar
  48. Stellwagen, E. (1977). Acc. Chem. Res., 10, 92CrossRefGoogle Scholar
  49. Stellwagen, E., Cass, R., Thompson, S. T. and Woody, M. (1975). Nature, Lond., 257, 716CrossRefGoogle Scholar
  50. Sulkowski, E. (1985). Trends Biotechnol., 3, 1CrossRefGoogle Scholar
  51. Thompson, S. T. and Stellwagen, E. (1976). Proc. Natl Acad. Sci. USA, 73, 361CrossRefGoogle Scholar
  52. Warner, R. C. (1954). In The Proteins (ed. H. Neurath and K. Bailey), Vol.2a, pp. 435–485, Academic Press, New YorkCrossRefGoogle Scholar
  53. Wilson, J. E. (1976). Biochem. Biophys. Res. Commun., 72, 816CrossRefGoogle Scholar
  54. Witt, J. J. and Roskoski, R. (1980). Biochemistry, 19, 143CrossRefGoogle Scholar
  55. Yon, R. J. (1977). Biochem. J., 161, 233CrossRefGoogle Scholar

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© The contributors 1987

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  • P. Hughes
  • R. F. Sherwood

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