Journal of Protein Chemistry

, Volume 14, Issue 6, pp 419–430 | Cite as

Protein selectivity in immobilized metal affinity chromatography based on the surface accessibility of aspartic and glutamic acid residues

  • Michael Zachariou
  • Milton T. W. Hearn
Article

Abstract

The interaction of different species variants of cytochrome c and myoglobin, as well as hen egg white lysozyme, with the hard Lewis metal ions Al3+, Ca2+, Fe3+, and Yb3+ and the borderline metal ion Cu2+, immobilized to iminodiacetic acid (IDA)-Sepharose CL-4B, has been investigated over the rangepH 5.5–8.0. With appropriately chosen buffer and metal ion conditions, these proteins can be bound to the immobilized M n +-IDA adsorbents via negatively charged amino acid residues accessible on the protein surface. For example, tuna heart cytochrome c, which lacks surface-accessible histidine residues, readily bound to the Fe3+-IDA adsorbent, while the other proteins also showed affinity toward immobilized Fe3+-IDA adsorbents when buffers containing 30 mM of imidazole were used. These studies document that protein selectivity can be achieved with hard-metalion immobilized metal ion affinity chromatography (IMAC) systems through the interaction of surfaceexposed aspartic and glutamic acid residues on the protein with the immobilized M n +-IDA complex. These investigations have also documented that the so-called soft or borderline immobilized metal ions such as the Cu2+-IDA adsorbent can also interact with surface-accessible aspartic and glutamic acid residues in a protein-dependent manner. A relationship is evident between the number and extent of clustering of the surfaceaccessible aspartic and glutamic acid residues and protein selectivity with these IMAC systems. The use of elution buffers which contain organic compound modifiers which replicate the carboxyl group moieties of these amino acids on the surface of proteins is also described.

Key words

Immobilized metal ion affinity chromatography cytochrome c surface accessibility hard metal ion chelates 

Abbreviations

IDA

iminodiacetic acid

IDA-Mn+

iminodiacetic acid chelated to metal ion

IMAC

immobilized metal affinity chromatography

DHCC

dog heart cytochrome c

HHCC

horse heart cytochrome c, THCC, tuna heart cytochrome c

HMYO

horse skeletal muscle myoglobin

SMYO

sheep skeletal muscle myoglobin

HEWL

hen egg white lysozyme

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Andersson, L. J. (1991).J. Chromatogr. 539, 327–334.CrossRefGoogle Scholar
  2. Andersson, L. J., and Porath, J. (1986).Anal. Biochem. 154, 250–254.CrossRefPubMedGoogle Scholar
  3. Andersson, L. J., Sulkowski, E., and Porath, J. (1991).Bioseparation 2, 15–22.PubMedGoogle Scholar
  4. Arnold, F. H. (1991).Bio Technology 9, 151–156.PubMedGoogle Scholar
  5. Bayes, C. F., and Mesmer, R. E. (1976).The Hydrolysis of Cations, Wiley, New York, pp. 120–135.Google Scholar
  6. Bollin, E., and Sulkowski, E. (1978).Arch. Virol. 58, 149–152.CrossRefPubMedGoogle Scholar
  7. Bushnell, G. D., Louie, G. V., and Brayer, G. D. (1990).J. Mol. Biol. 214, 585–595.PubMedGoogle Scholar
  8. Chaga, C., Andersson, L., Ersson, B., and Porath, J. (1989).Biotechnol. Appl. Biochem. 11, 424–431.PubMedGoogle Scholar
  9. Cohen, D., and Hayes, (1974).Google Scholar
  10. Dayhoff, M. O. (1976).Atlas of Protein Sequence and Structure, supplement 2, National Biomedical Research Foundation.Google Scholar
  11. Dickerson, R. E. (1972).Sci. Am. 226, 58–72.PubMedGoogle Scholar
  12. Dickerson, R. E., Takano, T., Eisenberg, D. Kallai, O. B., Samson, L., Cooper, A., and Margoliash, E. (1971).J. Biol. Chem. 246, 1511–1535.PubMedGoogle Scholar
  13. Everson, R. J., and Parker, H. E. (1974).Bioinorg. Chem. 4, 15–20.CrossRefPubMedGoogle Scholar
  14. Fatiadi, A. J. (1987).CRC Crit. Rev. Anal. Chem. 18, 1–44.Google Scholar
  15. Glusker, J. P. (1991).Adv. Protein Chem. 42, 1–76.PubMedGoogle Scholar
  16. Good, N. E., Winget, G. D., Winter, W., Connally, T. N., and Singh, R. N. N. (1966).Biochemistry 5, 467–474.PubMedGoogle Scholar
  17. Gurd, F. R. N., and Wilcox, P. E. (1956).Adv. Protein Chem. 11, 311–427.Google Scholar
  18. Helfferich, F. (1961).Nature 189, 1001–1005.Google Scholar
  19. Hemdan, E. S., Sulkowski, E., and Porath, J. (1989).Proc. Natl. Acad. Sci. USA 86, 1811–1815.PubMedGoogle Scholar
  20. Hochuli, E., Bannwarth, W., Dobeli, H., Gentz, R., and Stuber, D. (1988).Bio/Technology 6, 1321–1325.Google Scholar
  21. Hutchens, T. W., and Yip, T.-T. (1990).J. Chromatogr. 500, 531–542.PubMedGoogle Scholar
  22. Hutchens, T. W., and Yip, T.-T. (1991).J. Inorg. Biochem. 42, 105–118.PubMedGoogle Scholar
  23. Hutchens, T. W., Yip, T.-T., and Porath, J. (1988).Anal. Biochem. 170, 168–182.PubMedGoogle Scholar
  24. Kabasch, W., and Saunders, C. (1983).Biopolymers 22, 2577–2637.CrossRefGoogle Scholar
  25. Kagedal, L. (1989).Protein Purification (Janson, J. C., and Ryden, L., eds), VCH, New York, pp. 227–251.Google Scholar
  26. Mantovaara, T. (1990). The use of calcium(II) and cobalt(II) as adsorbents in immobilized metal affinity chromatography, Ph.D. Thesis, University of Uppsala, Uppsala.Google Scholar
  27. Meinhardt, J. E. (1948).Science 110, 387–395.Google Scholar
  28. Muszynska, G., Andersson, L. J., and Porath, J. (1986).Biochemistry 25, 6850–6853.PubMedGoogle Scholar
  29. Muszynska, G., Dobrowolska, G., Medin, A., Ekman, P., and Porath, J. (1992).J. Chromatogr. 604, 19–28.PubMedGoogle Scholar
  30. Neugebauer, J. (1988). A guide to the properties and uses of detergents in biology and biochemistry, Calbiochem.Google Scholar
  31. Pearson, R. G. (1990).Coord. Chem. Rev. 100, 403–425.Google Scholar
  32. Porath, J., and Olin, B. (1983).Biochemistry 22, 1621–1630.PubMedGoogle Scholar
  33. Porath, J., Carlsson, I., Olsson, I., and Belfrage, G. (1975).Nature 258, 598–599.PubMedGoogle Scholar
  34. Porath, J., Olin, B., and Granstrand, B. (1983).Arch. Biochem. Biophys. 225, 543–547.PubMedGoogle Scholar
  35. Ramadan, N., and Porath, J. (1985a).J. Chromatogr. 321, 93–104.PubMedGoogle Scholar
  36. Ramadan, N., and Porath, J. (1985b).J. Chromatogr. 321, 105–113.PubMedGoogle Scholar
  37. Richards, K. L., Aguilar, M. I., and Hearn, M. T. W. (1995).J. Chromatogr. 676, 17–31.Google Scholar
  38. Roos, P. H. (1991).J. Chromatogr. 587, 33–42.PubMedGoogle Scholar
  39. Schneider, W. (1984).Comments Inorg. Chem. 3, 205–223.Google Scholar
  40. Scopes, R. K. (1984).Anal. Biochem. 136, 525–529.PubMedGoogle Scholar
  41. Sillen, L. G., and Martell, A. E. (1971).Stability Constants, Supplement No. 1 (Special Publication No. 25), Chemical Society, London.Google Scholar
  42. Smith, M., Furman, T. C., and Pidgeon, C. (1987).Inorg. Chem. 26, 1965–1969.Google Scholar
  43. Smith, M., Furman, T. C., Ingolia, T. D., and Pidgeon, C. (1988).J. Biol. Chem. 263, 7211–7215.PubMedGoogle Scholar
  44. Sulkowski, E. (1985).Trends Biotechnol. 3, 1–5.Google Scholar
  45. Sulkowski, E. (1988).Makromol. Chem. Macromol. Symp. 17, 335–348.Google Scholar
  46. Sulkowski, E. (1989).Bioessays 10, 169–178.Google Scholar
  47. Takano, T., Kallai, O. B., Swanson, R., and Dickerson, R. E. (1973).J. Biol. Chem. 248, 5234–5255.PubMedGoogle Scholar
  48. Tarabe, S., Hiroyuki, N., and Ando, T. (1981).J. Chromatogr. 212, 295–304.PubMedGoogle Scholar
  49. Vijayalakshmi, M. A. (1983).Affinity Chromatography and Biological Recognition, Academic Press, New York, pp. 269–275.Google Scholar
  50. Zachariou, M., and Hearn, M. T. W. (1992).J. Chromatogr.,599, 171–177.Google Scholar
  51. Zachariou, M., Traverso, I., and Hearn, M. T. W. (1993).J. Chromatogr. 646, 107–120.PubMedGoogle Scholar
  52. Zhao, Y.-J., Sulkowski, E., and Porath, J. (1991).Eur. J. Biochem. 202, 1115–1119.PubMedGoogle Scholar

Copyright information

© Plenum Publishing Corporation 1995

Authors and Affiliations

  • Michael Zachariou
    • 1
  • Milton T. W. Hearn
    • 1
  1. 1.Centre for Bioprocess Technology, Department of Biochemistry and Molecular BiologyMonash UniversityClaytonAustralia

Personalised recommendations