title: Binding Studies of TNF Receptor Superfamily (TNFRSF) Receptors on Intact Cells.
abstract: Ligands of the tumor necrosis factor superfamily (TNFSF) interact with members of the TNF receptor superfamily (TNFRSF). TNFSF ligand-TNFRSF receptor interactions have been intensively evaluated by many groups. The affinities of TNFSF ligand-TNFRSF receptor interactions are highly dependent on the oligomerization state of the receptor, and cellular factors (e.g. actin cytoskeleton and lipid rafts) influence the assembly of ligand-receptor complexes, too. Binding studies on TNFSF ligand-TNFRSF receptor interactions were typically performed using cell-free assays with recombinant fusion proteins that contain varying numbers of TNFRSF ectodomains. It is therefore not surprising that affinities determined for an individual TNFSF ligand-TNFRSF interaction differ sometimes by several orders of magnitude and often do not reflect the ligand activity observed in cellular assays. To overcome the intrinsic limitations of cell-free binding studies and usage of recombinant receptor domains, we performed comprehensive binding studies with Gaussia princeps luciferase TNFSF ligand fusion proteins for cell-bound TNFRSF members on intact cells at 37 °C. The affinities of the TNFSF ligand G. princeps luciferase-fusion proteins ranged between 0.01 and 19 nm and offer the currently most comprehensive and best suited panel of affinities for in silico studies of ligand-receptor systems of the TNF family.
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Isabell
authors.initials: I
authors.lastname: Lang
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Füllsack
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Agnes
authors.initials: A
authors.lastname: Wyzgol
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Fick
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Trebing
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: José Antonio Carmona
authors.initials: JA
authors.lastname: Arana
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Viktoria
authors.initials: V
authors.lastname: Schäfer
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Weisenberger
authors.affiliation: From the Division of Molecular Internal Medicine, Department of Internal Medicine II, University Hospital Würzburg, Röntgenring 11, 97070 Würzburg, Germany harald.wajant@mail.uni-wuerzburg.de.
authors.forename: Harald
authors.initials: H
authors.lastname: Wajant
mesh_terms: D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000235:genetics; D011485:Protein Binding; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins
chemical_list: D016207:Cytokines; D018124:Receptors, Tumor Necrosis Factor; D011994:Recombinant Proteins; D008156:Luciferases
keywords: NF-κB (NF-KB); TNF receptor-associated factor; cytokine; fusion protein; ligand-binding protein; protein complex; receptor regulation; tumor necrosis factor (TNF)