Identification and super-resolution imaging of ligand-activated receptor dimers in live cells
Sci Rep. 2013-08-08; 3(1):
DOI: 10.1038/srep02387
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1. Sci Rep. 2013;3:2387. doi: 10.1038/srep02387.
Identification and super-resolution imaging of ligand-activated receptor dimers
in live cells.
Winckler P(1), Lartigue L, Giannone G, De Giorgi F, Ichas F, Sibarita JB, Lounis
B, Cognet L.
Author information:
(1)LP2N, University of Bordeaux, UMR 5298, F-33405 Talence, France.
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.
DOI: 10.1038/srep02387
PMCID: PMC3737505
PMID: 23925048 [Indexed for MEDLINE]