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Applications of Differential Scanning Fluorometry and Related Technologies in Characterization of Protein–Ligand Interactions

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Book cover Targeting Enzymes for Pharmaceutical Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2089))

Abstract

Differential scanning fluorometry (DSF) is an efficient and high-throughput method to analyze protein stability, as well as detect ligand interactions through perturbations of the protein’s melting temperature. The method monitors protein unfolding by observing the fluorescence changes of a sample, whether through an environmentally sensitive fluorophore or by intrinsic protein fluorescence, while a temperature gradient is applied. Here, we describe in detail how to develop and optimize DSF assays to identify protein–ligand interactions while exploring different buffer and additive conditions. Analysis of the data and further applications of the method are also discussed.

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Authors and Affiliations

  1. National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA

    Bolormaa Baljinnyam, Michael Ronzetti, Adam Yasgar & Anton Simeonov

Authors
  1. Bolormaa Baljinnyam
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  2. Michael Ronzetti
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  3. Adam Yasgar
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  4. Anton Simeonov
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Corresponding author

Correspondence to Anton Simeonov .

Editor information

Editors and Affiliations

  1. Department of Biotechnology, Agricultural University of Athens, Athens, Greece

    Nikolaos E. Labrou

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Baljinnyam, B., Ronzetti, M., Yasgar, A., Simeonov, A. (2020). Applications of Differential Scanning Fluorometry and Related Technologies in Characterization of Protein–Ligand Interactions. In: Labrou, N. (eds) Targeting Enzymes for Pharmaceutical Development. Methods in Molecular Biology, vol 2089. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0163-1_4

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  • DOI: https://doi.org/10.1007/978-1-0716-0163-1_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0162-4

  • Online ISBN: 978-1-0716-0163-1

  • eBook Packages: Springer Protocols

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