Fluorescence resonance energy transfer-based technologies in the study of protein-protein interactions at the cell surface.
Understanding of the molecular mechanisms of protein-protein interactions (PPIs) at the cell surface of living cells is fundamental to comprehend the functional meaning of a large number of cellular processes. Here we discuss how new methodological strategies derived from non-invasive fluorescence-based approaches (i.e. fluorescence resonance energy transfer, FRET) have been successfully developed to characterize plasma membrane PPIs. Importantly, these technologies alone - or in concert with complementary methods (i.e. SNAP-tag/TR-FRET, TIRF/FRET) - can become extremely powerful approaches for visualizing cell surface PPIs, even between more than two proteins and also in native tissues. Interestingly, these methods would also be relevant in drug discovery in order to develop new high-throughput screening approaches or to identify new therapeutic targets. Accordingly, herein we provide a thorough assessment on all biotechnological aspects, including strengths and weaknesses, of these fluorescence-based methodologies when applied in the study of PPIs occurring at the cell surface of living cells.
Unitat de Farmacologia, Departament de Patologia i Terapèutica Experimental, Facultat de Medicina, Universitat de Barcelona, L'Hospitalet de Llobregat, 08907 Barcelona, Spain.
Víctor
V
Fernández-Dueñas
Javier
J
Llorente
Jorge
J
Gandía
Dasiel O
DO
Borroto-Escuela
Luigi F
LF
Agnati
Carla I
CI
Tasca
Kjell
K
Fuxe
Francisco
F
Ciruela
D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods; D013194:Staining and Labeling
D005456:Fluorescent Dyes; D008565:Membrane Proteins