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

与食物摄入和癌症相关的神经肽 Y5 受体的结构解析

• 批准号: 468939089
• 负责人: Professorin Dr. Annette G. Beck-Sickinger
• 金额: --
• 依托单位: Arbeitsgruppe Biochemie und Bioorganische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/468939089?language=en
• 关键词: Structure; Elucidation; Neuropeptide; Y5; Receptor

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.

该项目的目的是阐明人类神经肽Y 5受体（NPY 5 R）的结构，该受体在食物摄入和癌症中起着重要作用。我们的目标是生产大量高纯度和热稳定的NPY 5 R样品，用于生物物理和功能研究。将设计、表达和纯化NPY 5 R构建体用于功能和结晶研究，并表征配体结合和活化。将尝试制备稳定的NPY 5 R-G蛋白/arrestin三元复合物的样品。将设计和应用可用于稳定NPY 5 R和探测配体-受体相互作用的化学工具。稳定GPCR的配体的可用性是成功确定其结构的关键因素。我们将使用我们和其他人开发的小分子和肽配体来研究相互作用。设计和生产的工具将包括荧光探针，建立基于nanolucerifase的结合测定。最后，我们将通过确定其3D结构来了解NPY 5 R配体结合特性和信号传导机制。稳定的受体和配体以及下游信号蛋白将用于cryo-EM（用于受体-下游信号蛋白复合物）和结晶（用于受体-配体）研究。一旦产生了结构，将进行进一步的研究，试图了解肽受体和受体信号蛋白的相互作用。受体的晶体结构的鉴定，如NPY 5 R将基本上有助于我们的理解以及特异性配体的开发。