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

Carrer, C., Stolz, M., Lewitzki, E., Rittmeyer, C., Kolbesen, B. O., & Grell, E. (2006). Removing coordinated metal ions from proteins: a fast and mild method in aequeous solution. Analytical and Bioanalytical Chemistry, 385, 1409-1413. doi:10.1007/s00216-006-0603-2.

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 Creators:
Carrer, Charlotte1, Author           
Stolz, Michael1, Author           
Lewitzki, Erwin2, Author           
Rittmeyer, Claudia, Author
Kolbesen, Bernd O., Author
Grell, Ernst1, Author           
Affiliations:
1Department of Biophysical Chemistry, Max Planck Institute of Biophysics, Max Planck Society, ou_2068289              
2Department of Molecular Neurogenetics, Max Planck Institute of Biophysics, Max Planck Society, ou_2068296              

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 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.

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Language(s): eng - English
 Dates: 2006-07-042006-08
 Publication Status: Published in print
 Pages: 6
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 305373
DOI: 10.1007/s00216-006-0603-2
 Degree: -

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Title: Analytical and Bioanalytical Chemistry
  Abbreviation : Anal. Bioanal. Chem.
Source Genre: Journal
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Publ. Info: Heidelberg : Springer-Verlag
Pages: - Volume / Issue: 385 Sequence Number: - Start / End Page: 1409 - 1413 Identifier: ISSN: 1618-2642
CoNE: https://pure.mpg.de/cone/journals/resource/111006469468428