Structural studies interaction between CRF G-protein co

促肾上腺皮质激素释放因子（CRF）及其相关配体不仅在调节内分泌、中枢神经和免疫系统对应激的适应性反应中起关键作用，而且在心血管和胃肠道系统中也起关键作用。配体的作用通过与CRF G蛋白偶联受体（GPCR）结合来介导。本申请的目的是CRF家族的肽与其受体之间的相互作用的结构阐明。拟议的研究将利用激动剂/拮抗剂-受体相互作用的特异性，将阐明通过膜的信号传导途径的分子机制，并将促进针对应激相关疾病和其他疾病的药物的改进。对上述目的的重要观察是确定了 CRF肽家族，其位于CRFR的N-末端胞外结构域（ECD 1）。我们确定了CRFR 2 β胞外结构域的三维NMR结构，并确定了其配体结合位点。在这里，我们建议阐明的动态和三维结构的细胞外CRF受体的自由和复杂的激动剂和拮抗剂。此外，提出了通过转移的NOE和“转移的”残余偶极偶联来确定激素类似物与全长受体结合时的三维结构，并且平行地，旨在使用我们建立的功能性表达方案获得全长受体的结构见解。 CRFR 1在E. coli和TROSY-NMR。我们相信，这些研究将提供配体-受体特异性的分子见解，并为改善针对应激相关疾病的药物奠定结构基础。将使用标准协议确定结构和动力学。

Corticotropin releasing factor (CRF) and related ligands play key not only roles in the modulation of adaptive responses of the endocrine, central nervous and immune system to stress but also within the cardiovascular and gastrointestinal systems. The ligand's effects are mediated by binding to CRF G-protein coupled receptors (GPCR's). The aim of this application is the structural elucidation of the interaction between peptides of the CRF family and their receptors. The proposed studies will exploit the specificities of agonist/antagonist-receptor interactions, will elucidate the molecular mechanism of the signaling pathway through the membrane and will facilitate the improvement of drugs targeting stress-related and other diseases. The important observation towards the aims described above is the determination of a major binding site of the CRF family of peptides, which is located in the N-terminal extracellular domain (ECD1) of CRFR's. We determined the three-dimensional NMR structure of the extracellular domain of CRFR2beta and identified its ligand binding site. Here, we propose to elucidate the dynamics and the three-dimensional structures of the extracellular CRF receptors both free and in complex with agonists and antagonists. It is furthermore proposed to determine the three dimensional structure of a hormone analogue when bound to full-length receptor through transferred NOEs and "transferred" residual dipolar couplings, and in parallel, it is aimed to obtain structural insights of the full-length receptor using our established expression protocol of functional CRFR1 in E. coli and TROSY-NMR. We believe that these studies will provide molecular insights of the ligand-receptor specificities and set a structural base for the improvement of drugs targeting stress-related diseases. The structures and dynamics will be determined using standard protocols.