Structural studies interaction between CRF G-protein coupled receptors & ligands

CRF G 蛋白偶联受体之间相互作用的结构研究

• 批准号: 7864288
• 负责人: ROLAND P RIEK
• 金额: $ 17.91万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7864288
• 关键词: Affinity; Agonist; Amino Acids; Binding; Binding Sites; Biology; CRF receptor type 2; Cardiovascular system; Complex; Corticotropin-Releasing Hormone; Corticotropin-Releasing Hormone Receptors; Cyclic AMP; Disease; Drug Delivery Systems; ERBB2 gene; Endocrine; Escherichia coli; Extracellular Domain; Family; G-Protein-Coupled Receptors; Goals; Hormone Receptor; Immune system; Investigation; Knowledge; Label; Length; Ligand Binding; Ligands; Mammals; Measurement; Measures; Mediating; Membrane; Methods; Molecular; Molecular Conformation; N-terminal; Neurosecretory Systems; Peptide Hormones Receptors; Peptides; Pharmaceutical Preparations; Play; Proteins; Protocols documentation; Receptor Activation; Relative (related person); Relaxation; Research Personnel; Residual state; Role; Signal Pathway; Specificity; Stress; Structure; Transmembrane Domain; analog; astressin; base; corticotropin releasing factor-binding protein; extracellular; gastrointestinal system; hormone analog; insight; interest; member; novel; peptide hormone; programs; protein protein interaction; receptor; research study; response; restraint; three dimensional structure; urocortin

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.

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系统中也起着关键作用。该配体的作用是通过与CRF G蛋白偶联受体(GPCRs)结合来实现的。本应用的目的是从结构上阐明CRF家族多肽与其受体之间的相互作用。这些研究将利用激动剂/拮抗剂-受体相互作用的特殊性，阐明通过膜的信号通路的分子机制，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一个主要的结合部位 CRF家族多肽，位于CRFR的N端胞外区(ECD1)。我们测定了CRFR2β胞外区的三维核磁共振结构，并确定了其配体结合部位。在这里，我们建议阐明细胞外CRF受体的动力学和三维结构，既有游离的，也有与激动剂和拮抗剂复合的。此外，还建议确定激素类似物通过转移的noes和“转移的”残余偶极偶联与全长受体结合时的三维结构，并旨在利用我们建立的功能性的表达方案获得全长受体的结构洞察力。 在大肠杆菌中表达CRFR1，并进行TROSY-核磁共振。我们相信，这些研究将提供配体-受体特异性的分子洞察力，并为改进针对应激相关疾病的药物奠定结构基础。结构和动力学将使用标准协议来确定。