title: Cooperativity in the interaction of synthetic CD40L mimetics with CD40 and its implication in cell signaling.
abstract: CD40 ligand (CD40L) and CD40 are members of the tumor necrosis factor (TNF) and TNF receptor superfamilies, respectively. Their interaction is crucial for the development of a proper immune response. Intervention on this pathway provides an important ground for new treatments targeting autoimmune diseases or helping to fight infection and cancer. We have recently reported on the structure-based design of synthetic molecules with C3 symmetry, named mini-CD40Ls, that can effectively mimic homotrimeric soluble CD40L. Here we show that substitution of a D-prolyl residue for the glycyl within the Lys-Gly-Tyr-Tyr CD40-binding motif leads to a complete loss of cooperativity in the interaction of the mimetic with its cognate receptor as assessed by surface plasmon resonance experiments. The ability of the modified mini-CD40L to induce apoptosis on both human and murine lymphoma cells was not affected by this mutation. However, it was unable to induce the NF-kappaB pathway in the mouse D1 dendritic cell line, which is essential for its complete maturation, but still activated production of IL-12 p40 mRNA. These differential effects might be partly explained by the change in rigidity of the CD40 recognition element. In this study, we not only point out the consequences of the abrogation of the cooperative property in a ligand-receptor interaction on downstream cellular events but also demonstrate the usefulness of synthetic multivalent ligands in dissecting the complex mechanisms implicated in the signalosome.
authors.affiliation: CNRS UPR 9021, Immunologie et Chimie Thérapeutiques, Institut de Biologie Moléculaire et Cellulaire, 15 rue René Descartes, 67084 Strasbourg, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Wieckowski
authors.forename: Nathalie
authors.initials: N
authors.lastname: Trouche
authors.forename: Olivier
authors.initials: O
authors.lastname: Chaloin
authors.forename: Gilles
authors.initials: G
authors.lastname: Guichard
authors.forename: Sylvie
authors.initials: S
authors.lastname: Fournel
authors.forename: Johan
authors.initials: J
authors.lastname: Hoebeke
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D017209:Apoptosis; D019013:CD40 Antigens / Q000235:genetics / Q000378:metabolism*; D023201:CD40 Ligand / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003713:Dendritic Cells / Q000276:immunology; D005998:Glycine / Q000502:physiology; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D011392:Proline / Q000502:physiology; D015398:Signal Transduction; D020349:Surface Plasmon Resonance
chemical_list: D019013:CD40 Antigens; D023201:CD40 Ligand; D011392:Proline; D005998:Glycine