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Receptor–Ligand Interactions: Binding Affinities Studied by Single-Molecule and Super-Resolution Microscopy on Intact Cells

Authors

  • Marina S. Dietz,

    1. Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe University, Max-von-Laue-Str. 7, 60438 Frankfurt (Germany)
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  • Franziska Fricke,

    1. Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe University, Max-von-Laue-Str. 7, 60438 Frankfurt (Germany)
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  • Carmen L. Krüger,

    1. Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe University, Max-von-Laue-Str. 7, 60438 Frankfurt (Germany)
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  • Prof. Dr. Hartmut H. Niemann,

    1. Department of Chemistry, Bielefeld University, Universitätsstr. 25, 33615 Bielefeld (Germany)
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  • Prof. Dr. Mike Heilemann

    Corresponding author
    1. Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe University, Max-von-Laue-Str. 7, 60438 Frankfurt (Germany)
    • Institute of Physical and Theoretical Chemistry, Johann Wolfgang Goethe University, Max-von-Laue-Str. 7, 60438 Frankfurt (Germany)

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Abstract

Protein–ligand interactions play an important role in many biological processes. Notably, membrane receptors are the starting point for a huge variety of cellular signal transduction pathways. Quantifying the binding affinity of a ligand for its transmembrane receptor is of great importance as it provides information on the potency of the ligand. We developed a new experimental procedure to determine binding affinities of ligands for their membrane receptors directly on intact single cells using super-resolution imaging. Dissociation constants were determined by titrating fluorophore-labelled ligand against cells expressing the target protein and applying single-molecule imaging.

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