Receptor-ligand interactions: binding affinities studied by single-molecule and super-resolution microscopy on intact cells.
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.
Marina S
MS
Dietz
Franziska
F
Fricke
Carmen L
CL
Krüger
Hartmut H
HH
Niemann
Mike
M
Heilemann
D001665:Binding Sites; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008853:Microscopy / Q000379:methods*; D018160:Receptors, Cytoplasmic and Nuclear / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D008024:Ligands; D018160:Receptors, Cytoplasmic and Nuclear