Structure Elucidation of the Neuropeptide Y5 Receptor- Relevant in Food intake and Cancer

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The aim of the project is to elucidate the structure of the human neuropeptide Y5 receptor (NPY5R), which plays an important role in food intake and cancer. We aim to produce large samples of highly purified and thermally stabilized NPY5R for biophysical and functional studies. NPY5R constructs will be designed, expressed and purified for functional and crystallization studies and characterized for ligand binding and activation. Attempts will be made to prepare samples of stabilized NPY5R-G protein/arrestin ternary complexes.Chemical tools that can be used to stabilize NPY5R and to probe ligand-receptor interactions will be designed and applied.The availability of ligands that stabilize GPCRs has been a critical factor in the successful determination of their structures. We will use small molecule and peptide ligands that have been developed by us and others for studying interactions.Tools designed and produced will include fluorescence probes, the set-up of a nanolucerifase based binding assay. Finally, we will develop an understanding of NPY5R ligand binding properties and signalling mechanism by determining its 3D structure. Stabilized receptors and ligands as well as downstream signalling proteins, will be used in cryo-EM (for receptor-downstream signalling protein complexes) and crystallization (for receptor-ligand) studies. Once a structure has been generated, further studies will be conducted to attempt to understand peptide-receptor and receptor-signalling protein interactions.The identification of the crystal structure of receptors, such as the NPY5R will essentially contribute to our understanding as well as the development of specific ligands.