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Analyzing Transmembrane Protein and Hydrophobic Helix Topography by Dual Fluorescence Quenching

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Lipid-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 974))

Abstract

The location of fluorescent groups relative to the lipid bilayer can be evaluated using fluorescence quenchers embedded in the membrane and/or dissolved in aqueous solution. Quenching can be used to define the membrane topography of membrane proteins and individual membrane-embedded hydrophobic helices by combining it with the placement of fluorescent groups, including Trp, at defined sequence positions. This chapter briefly discusses various quenching methods for studies of membrane protein topography and provides detailed protocols for dual quencher analysis, a rapid, highly sensitive, and experimentally flexible approach in which the information gained from both a membrane-embedded and aqueous quencher is combined.

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Acknowledgments

This work was supported by NSF grant MCB 1019986 to E.L.

Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Rowan University, Glassboro, NJ, USA

    Gregory A. Caputo

  2. Department of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, NY, USA

    Erwin London

Authors
  1. Gregory A. Caputo
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  2. Erwin London
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Corresponding author

Correspondence to Erwin London .

Editor information

Editors and Affiliations

  1. Institut für Biologie, Abteilung Biophysik, Universität Kassel, Heinrich-Plett-Straße 40, Kassel, 34132, Germany

    Jörg H. Kleinschmidt

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Caputo, G.A., London, E. (2013). Analyzing Transmembrane Protein and Hydrophobic Helix Topography by Dual Fluorescence Quenching. In: Kleinschmidt, J. (eds) Lipid-Protein Interactions. Methods in Molecular Biology, vol 974. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-275-9_13

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  • DOI: https://doi.org/10.1007/978-1-62703-275-9_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-274-2

  • Online ISBN: 978-1-62703-275-9

  • eBook Packages: Springer Protocols

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