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Cytochrome P450 Protocols pp 31–38Cite as

Purification of Cytochromes P450

Products of Bacterial Recombinant Expression Systems

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 320))

Abstract

A general procedure for the solubilization of cytochrome P450 (P450) from bacterial membranes specifically for a human P450 expressed heterologously in the host Escherichia coli is described. The example involves the use of a P450 (3A4) with a C-terminal oligohistidine tag and includes sequential DEAE and metal affinity chromatography.

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Acknowledgments

We thank N. A. Hosea for helping to develop and implement the particular strategy used here, and W. A. McCormick for comments on the manuscript. This work was supported in part by USPHS grants R01 CA90426 and P30 ES00267.

Author information

Authors and Affiliations

  1. Department of Biochemistry, Center in Molecular Toxicology, Vanderbilt University School of Medicine, Nashville, TN, USA

    F. Peter Guengerich & Martha V. Martin

Authors
  1. F. Peter Guengerich
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  2. Martha V. Martin
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Editor information

Editors and Affiliations

  1. School of Biological Sciences, Queen Mary University of London, London, UK

    Ian R. Phillips

  2. Department of Biochemistry and Molecular Biology, University College London, London, UK

    Elizabeth A. Shephard

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© 2006 Humana Press Inc.

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Guengerich, F.P., Martin, M.V. (2006). Purification of Cytochromes P450. In: Phillips, I.R., Shephard, E.A. (eds) Cytochrome P450 Protocols. Methods in Molecular Biology, vol 320. Humana Press, Totowa, NJ. https://doi.org/10.1385/1-59259-998-2:31

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  • DOI: https://doi.org/10.1385/1-59259-998-2:31

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-58829-441-8

  • Online ISBN: 978-1-59259-998-1

  • eBook Packages: Springer Protocols

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