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Affinity Purification of Membrane Proteins

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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

Biochemical, biophysical, and structural studies of membrane proteins rely on the availability of highly pure and monodisperse membrane protein samples. One of the most powerful methods for isolation of the membrane protein of interest is affinity purification. This methodology typically relies on engineering an affinity tag into the protein of interest and an affinity resin that specifically recognizes the tag, allowing one to purify the target protein in a single step. In some cases, the affinity purification procedure is combined with additional steps to increase the purity and homogeneity of the final protein sample. Here, we describe several protocols for affinity purification of TSPO, a small membrane protein. The techniques we use include immobilized metal affinity chromatography (IMAC) and strep-II tag-based streptavidin affinity chromatography.

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References

  1. Seddon AM, Curnow P, Booth PJ (2004) Membrane proteins, lipids and detergents: not just a soap opera. Biochim Biophys Acta Biomembr 1666(1):105–117. https://doi.org/10.1016/j.bbamem.2004.04.011

    Article  CAS  Google Scholar 

  2. Schmidt TGM, Skerra A (2007) The Strep-tag system for one-step purification and high-affinity detection or capturing of proteins. Nat Protoc 2:1528. https://doi.org/10.1038/nprot.2007.209

    Article  CAS  PubMed  Google Scholar 

  3. Kimple ME, Brill AL, Pasker RL (2013) Overview of Affinity Tags for Protein Purification. Current protocols in protein science / editorial board, John E Coligan [et al] 73:Unit-9.9. doi:https://doi.org/10.1002/0471140864.ps0909s73

  4. Hanson MA, Cherezov V, Griffith MT, Roth CB, Jaakola V-P, Chien EYT, Velasquez J, Kuhn P, Stevens RC (2008) A Specific Cholesterol Binding Site Is Established by the 2.8 Å Structure of the Human β2-Adrenergic Receptor. Structure 16(6):897–905. https://doi.org/10.1016/j.str.2008.05.001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Garavito RM, Ferguson-Miller S (2001) Detergents as Tools in Membrane Biochemistry. J Biol Chem 276(35):32403–32406. https://doi.org/10.1074/jbc.R100031200

    Article  CAS  Google Scholar 

  6. Privé GG (2007) Detergents for the stabilization and crystallization of membrane proteins. Methods 41(4):388–397. https://doi.org/10.1016/j.ymeth.2007.01.007

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Volodymyr M. Korkhov .

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Graeber, E., Korkhov, V.M. (2020). Affinity Purification of Membrane Proteins. 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_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0373-4_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0372-7

  • Online ISBN: 978-1-0716-0373-4

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