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Membrane Protein Solubilization and Quality Control: An Example of a Primary Active Transporter

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Expression, Purification, and Structural Biology of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2127))

Abstract

When purifying a membrane protein, finding a detergent for solubilization is one of the first steps to master. Ideally, only little time is invested to identify the best-suited detergent, which on the one hand would solubilize large amounts of the target protein but on the other hand would sustain the protein’s activity. Here we describe the solubilization screen and subsequent activity assay we have optimized for the bacterial P-type ATPase KdpFABC. In just 2 days, more than 70 detergents were tested for their solubilization potential. Afterwards, a smaller selection of the successful detergents was assayed for their ability to retain the activity of the membrane protein complex.

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Authors and Affiliations

  1. Institute of Biochemistry, Biocenter, Goethe University Frankfurt, Frankfurt, Germany

    Charlott Stock & Inga Hänelt

Authors
  1. Charlott Stock
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  2. Inga Hänelt
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Corresponding author

Correspondence to Inga Hänelt .

Editor information

Editors and Affiliations

  1. Biozentrum, University of Basel, Basel, Basel Stadt, Switzerland

    Camilo Perez

  2. Biozentrum, University of Basel, Basel, Basel Stadt, Switzerland

    Timm Maier

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Stock, C., Hänelt, I. (2020). Membrane Protein Solubilization and Quality Control: An Example of a Primary Active Transporter. In: Perez, C., Maier, T. (eds) Expression, Purification, and Structural Biology of Membrane Proteins. Methods in Molecular Biology, vol 2127. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0373-4_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0373-4_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0372-7

  • Online ISBN: 978-1-0716-0373-4

  • eBook Packages: Springer Protocols

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