Identification and super-resolution imaging of ligand-activated receptor dimers in live cells

Abstract : Molecular interactions are key to many chemical and biological processes like protein function. In many signaling processes they occur in sub-cellular areas displaying nanoscale organizations and involving molecular assemblies. The nanometric dimensions and the dynamic nature of the interactions make their investigations complex in live cells. While super-resolution fluorescence microscopies offer live-cell molecular imaging with sub-wavelength resolutions, they lack specificity for distinguishing interacting molecule populations. Here we combine super-resolution microscopy and single-molecule Förster Resonance Energy Transfer (FRET) to identify dimers of receptors induced by ligand binding and provide super-resolved images of their membrane distribution in live cells. By developing a two-color universal-Point-Accumulation-In-the-Nanoscale-Topography (uPAINT) method, dimers of epidermal growth factor receptors (EGFR) activated by EGF are studied at ultra-high densities, revealing preferential cell-edge sub-localization. This methodology which is specifically devoted to the study of molecules in interaction, may find other applications in biological systems where understanding of molecular organization is crucial.
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Scientific Reports, Nature Publishing Group, 2013, 3, pp.2387. 〈10.1038/srep02387〉
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Pascale Winckler, Lydia Lartigue, Gregory Giannone, Francesca De Giorgi, François Ichas, et al.. Identification and super-resolution imaging of ligand-activated receptor dimers in live cells. Scientific Reports, Nature Publishing Group, 2013, 3, pp.2387. 〈10.1038/srep02387〉. 〈hal-00909281〉

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