Skip to main content
SpringerLink
Log in

Removing coordinated metal ions from proteins: a fast and mild method in aqueous solution

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Thermodynamic and kinetic studies of metal binding to proteins require the investigation of metal-free proteins, which are often difficult to obtain. We have developed a very fast and mild method to eliminate metal ions from proteins by column chromatography using a commercially available Ni-NTA-type stationary phase. This material, initially designed for protein purification purposes in biotechnology, acts as a strong cation chelator when Ni2+ ions are removed. We have tested this new method with Ca-ATPase, an integral membrane protein exhibiting a strong affinity for Ca2+. By eluting the protein over the Ni2+-free NTA gel, we could remove 95% of the total Ca2+ and obtain an essentially Ca2+-free protein. This method is efficient with only a small amount of NTA gel, and we suggest that it can be applied in general for removal of metal ions from proteins. Moreover, as this procedure can be carried out under mild conditions, the chosen protein kept its enzymatic activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. Mun S, Chin C, Xie Y, Wang NHL (2006) Ind Eng Chem Res 45:316–329

    Article  CAS  Google Scholar 

  2. Porath J, Carlsson J, Olson I, Belfrage G (1975) Nature 258:598–599

    Article  CAS  Google Scholar 

  3. Hochuli E, Dobeli H, Schacher A (1987) J Chromatog 411:177–184

    Article  CAS  Google Scholar 

  4. Kim YM, Ogawa W, Tamai E, Kuroda T, Mizushima T, Tsuchiya T (2002) J Biochem 132:71–76

    CAS  Google Scholar 

  5. De Meis L, Hasselbach W (1971) J Biol Chem 246:4759–4763

    Google Scholar 

  6. Champeil P, Guillain F, Venien C, Gingold MP (1985) Biochemistry 24:69–81

    Article  CAS  Google Scholar 

  7. Meissner G, Conner GE, Fleischer S (1973) Biochim Biophys Acta 298:246–269

    Article  CAS  Google Scholar 

  8. Henderson MJ, Khan YM, East JM, Lee AG (1994) Biochem J 297:615–624

    CAS  Google Scholar 

  9. Lowry OH, Rosenbrough NJ (1951) J Biol Chem 193:265–275

    CAS  Google Scholar 

  10. Bradford M (1976) Anal Biochem 72:248–254

    Article  CAS  Google Scholar 

  11. Spector T (1978) Anal Biochem 86:142–146

    Article  CAS  Google Scholar 

  12. Petithory JR, Jencks WP (1988) Biochemistry 27:8626–8635

    Article  CAS  Google Scholar 

  13. Grynkiewicz G, Poenie M, Tsien RY (1985) J Biol Chem 260:3440–3450 and citations therein

    CAS  Google Scholar 

  14. Klockenkämper R (1997) Total-reflection X-ray fluorescence analysis. Wiley-Interscience, New York

    Google Scholar 

  15. Wittershagen A, Rostam-Khani P, Zickermann V, Zickermann I, Gemeinhardt S, Ludwig B, Kolbesen BO (1998) Fresenius J Anal Chem 361:326–328

    Article  CAS  Google Scholar 

  16. Orlowski S, Champeil P (1991) Biochemistry 30:352–361

    Article  CAS  Google Scholar 

  17. Peinelt C, Apell HJ (2002) Biophys J 82:170–181

    Article  CAS  Google Scholar 

  18. Martell AE, Smith RM (1974) Critical Stability Constants 1:139–144

    Google Scholar 

Download references

Acknowledgements

We wish to thank A. Schacht and H. Volk for valuable support and A.M. Albrecht-Gary for interesting discussions.

Author information

Authors and Affiliations

  1. Max-Planck-Institut für Biophysik, Max-von-Laue-Str. 3, 60438, Frankfurt am Main, Germany

    Charlotte Carrer, Michael Stolz, Erwin Lewitzki & Ernst Grell

  2. Institut für Anorganische und Analytische Chemie, Johann Wolfgang Goethe-Universität Frankfurt/Main, Marie-Curie-Str. 11, 60439, Frankfurt am Main, Germany

    Claudia Rittmeyer & Bernd O. Kolbesen

Authors
  1. Charlotte Carrer
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael Stolz
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Erwin Lewitzki
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Claudia Rittmeyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bernd O. Kolbesen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ernst Grell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ernst Grell.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Carrer, C., Stolz, M., Lewitzki, E. et al. Removing coordinated metal ions from proteins: a fast and mild method in aqueous solution. Anal Bioanal Chem 385, 1409–1413 (2006). https://doi.org/10.1007/s00216-006-0603-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-006-0603-2

Keywords

Search

Navigation