title: Quantitative single-molecule localization microscopy combined with rule-based modeling reveals ligand-induced TNF-R1 reorganization toward higher-order oligomers.
abstract: We report on the assembly of tumor necrosis factor receptor 1 (TNF-R1) prior to ligand activation and its ligand-induced reorganization at the cell membrane. We apply single-molecule localization microscopy to obtain quantitative information on receptor cluster sizes and copy numbers. Our data suggest a dimeric pre-assembly of TNF-R1, as well as receptor reorganization toward higher oligomeric states with stable populations comprising three to six TNF-R1. Our experimental results directly serve as input parameters for computational modeling of the ligand-receptor interaction. Simulations corroborate the experimental finding of higher-order oligomeric states. This work is a first demonstration how quantitative, super-resolution and advanced microscopy can be used for systems biology approaches at the single-molecule and single-cell level.
authors: Institute for Physical and Theoretical Chemistry, Johann-Wolfgang-Goethe-University Frankfurt, Frankfurt am Main, Germany.
authors: Franziska
authors: F
authors: Fricke
authors: Sebastian
authors: S
authors: Malkusch
authors: Gaby
authors: G
authors: Wangorsch
authors: Johannes F
authors: JF
authors: Greiner
authors: Barbara
authors: B
authors: Kaltschmidt
authors: Christian
authors: C
authors: Kaltschmidt
authors: Darius
authors: D
authors: Widera
authors: Thomas
authors: T
authors: Dandekar
authors: Mike
authors: M
authors: Heilemann
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D057054:Molecular Imaging / Q000379:methods*; D055503:Protein Multimerization / Q000187:drug effects*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*
chemical_list: D008024:Ligands; D047888:Receptors, Tumor Necrosis Factor, Type I; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha