title: G protein-coupled receptors sense fluid shear stress in endothelial cells.
abstract: Hemodynamic shear stress stimulates a number of intracellular events that both regulate vessel structure and influence development of vascular pathologies. The precise molecular mechanisms by which endothelial cells transduce this mechanical stimulus into intracellular biochemical response have not been established. Here, we show that mechanical perturbation of the plasma membrane leads to ligand-independent conformational transitions in a G protein-coupled receptor (GPCR). By using time-resolved fluorescence microscopy and GPCR conformation-sensitive FRET we found that stimulation of endothelial cells with fluid shear stress, hypotonic stress, or membrane fluidizing agent leads to a significant increase in activity of bradykinin B2 GPCR in endothelial cells. The GPCR conformational dynamics was detected by monitoring redistribution of GPCRs between inactive and active conformations in a single endothelial cell under fluid shear stress in real time. We show that this response can be blocked by a B(2)-selective antagonist. Our data demonstrate that changes in cell membrane tension and membrane fluidity affect conformational dynamics of GPCRs. Therefore, we suggest that GPCRs are involved in mediating primary mechanochemical signal transduction in endothelial cells. We anticipate our experiments to be a starting point for more sophisticated studies of the effects of changes in lipid bilayer environment on GPCR conformational dynamics. Furthermore, because GPCRs are a major target of drug development, a detailed characterization of mechanochemical signaling via the GPCR pathway will be relevant for the development of new antiatherosclerosis drugs.
authors.affiliation: La Jolla Bioengineering Institute, 505 Coast Boulevard South, La Jolla, CA 92037, USA. mirianas@ljbi.org
authors.forename: Mirianas
authors.initials: M
authors.lastname: Chachisvilis
authors.forename: Yan-Liang
authors.initials: YL
authors.lastname: Zhang
authors.forename: John A
authors.initials: JA
authors.lastname: Frangos
mesh_terms: D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D007038:Hypotonic Solutions; D008024:Ligands; D040542:Mechanotransduction, Cellular; D011487:Protein Conformation; D043782:Receptor, Bradykinin B2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D033081:Shear Strength; D013314:Stress, Mechanical
chemical_list: D007038:Hypotonic Solutions; D008024:Ligands; D043782:Receptor, Bradykinin B2; D011993:Recombinant Fusion Proteins