Structural studies of the human Notch-1 ligand-binding region. (360G-Wellcome-078765_Z_05_Z)

During development it is essential that cells are guided to their correct location within the embryo and that they differentiate into the appropriate cell type. A key short-range signal which has been highly conserved during metazoan evolution involves the Notch receptor which regulates spatial patterning, timing and outcomes of numerous cell fate decisions. A structural understanding of the Notch receptor -ligand interaction, which is currently lacking, would greatly enhance our understanding of the signalling mechanism and may lead to the design of novel agonists and antagonists of the receptor. With the current interest in cell-based therapies, manipulation of the Notch signal represents a promising strategy to produce defined cell types in vitro for therapeutic use. In this application we aim to i) crystallise and determine the individual atomic structures of the ligand binding region of human Notch-1 (hN-1) and the receptor binding portions of its ligands human Delta-like 1 and human Jagged-1 ii) identify the structural basis for the observed inhibitory effect of hN-1 EGF10 on ligand binding and the mechanism by which post-translational modification overcomes this effect iii) identify the stability of a Notch-ligand complex by SPR, and determine its atomic structure by X-ray crystallography and/or NMR shift mapping.