title: Fully quantified spectral imaging reveals in vivo membrane protein interactions.
abstract: Here we introduce the fully quantified spectral imaging (FSI) method as a new tool to probe the stoichiometry and stability of protein complexes in biological membranes. The FSI method yields two dimensional membrane concentrations and FRET efficiencies in native plasma membranes. It can be used to characterize the association of membrane proteins: to differentiate between monomers, dimers, or oligomers, to produce binding (association) curves, and to measure the free energies of association in the membrane. We use the FSI method to study the lateral interactions of Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), a member of the receptor tyrosine kinase (RTK) superfamily, in plasma membranes, in vivo. The knowledge gained through the use of the new method challenges the current understanding of VEGFR2 signaling.
authors.affiliation: Program in Molecular Biophysics, Johns Hopkins University, Baltimore, MD 21212, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: King
authors.forename: Michael
authors.initials: M
authors.lastname: Stoneman
authors.forename: Valerica
authors.initials: V
authors.lastname: Raicu
authors.forename: Kalina
authors.initials: K
authors.lastname: Hristova
mesh_terms: D000970:Antineoplastic Agents / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D015233:Models, Statistical; D009995:Osmosis; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism*; D015398:Signal Transduction; D013050:Spectrometry, Fluorescence / Q000379:methods; D013816:Thermodynamics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D005456:Fluorescent Dyes; D011506:Proteins; C000588866:KDR protein, human; D020794:Receptor Protein-Tyrosine Kinases; D040301:Vascular Endothelial Growth Factor Receptor-2