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Identification of Lipid Binding Modulators Using the Protein-Lipid Overlay Assay

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Proteomics for Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1647))

Abstract

The protein-lipid overlay assay is an inexpensive, easy-to-implement, and high-throughput methodology that employs nitrocellulose membranes to immobilize lipids in order to rapid screen and identify protein-lipid interactions. In this chapter, we show how this methodology can identify potential modulators of protein-lipid interactions by screening water-soluble lipid competitors or even the introduction of pH changes during the binding assay to identify pH-dependent lipid binding events.

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Acknowledgments

We thank Janet Webster for critical reading and comments on the manuscript. We also thank Tiffany Radle and Morgan Vaughn for the optimization of the purification conditions of the GST-EEA1 FYVE domain. Work in the Capelluto laboratory is supported by the National Institutes of Health (NIAID).

Author information

Authors and Affiliations

  1. Protein Signaling Domains Laboratory, Department of Biological Sciences, Biocomplexity Institute, Virginia Tech, Blacksburg, VA, 24061, USA

    Tuo-Xian Tang, Wen Xiong & Daniel G. S. Capelluto

  2. Center for Soft Matter and Biological Physics, Virginia Tech, Blacksburg, VA, 24061, USA

    Tuo-Xian Tang, Wen Xiong & Daniel G. S. Capelluto

  3. Integrated Cellular Responses Laboratory, Department of Biological Sciences, Biocomplexity Institute, Virginia Tech, Blacksburg, VA, 24061, USA

    Carla V. Finkielstein

Authors
  1. Tuo-Xian Tang
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  2. Wen Xiong
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  3. Carla V. Finkielstein
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  4. Daniel G. S. Capelluto
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Corresponding author

Correspondence to Daniel G. S. Capelluto .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, Virginia Tech, Blacksburg, Virginia, USA

    Iulia M. Lazar

  2. Department of Pharmaceutical Sciences, School of Pharmacy, Southern Illinois University Edwardsville, Edwardsville, Illinois, USA

    Maria Kontoyianni

  3. ImmunoGen, Waltham, Massachusetts, USA

    Alexandru C. Lazar

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Tang, TX., Xiong, W., Finkielstein, C.V., Capelluto, D.G.S. (2017). Identification of Lipid Binding Modulators Using the Protein-Lipid Overlay Assay. In: Lazar, I., Kontoyianni, M., Lazar, A. (eds) Proteomics for Drug Discovery. Methods in Molecular Biology, vol 1647. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7201-2_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7201-2_13

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7200-5

  • Online ISBN: 978-1-4939-7201-2

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