Molecular Mechanism of the NFkappaB Essential Modulator (NEMO) Scaffold Protein Mutated in Human Immunodeficiencies

人类免疫缺陷中 NFkappaB 必需调节剂 (NEMO) 支架蛋白突变的分子机制

• 批准号: 9177304
• 负责人: Adrian Whitty
• 金额: $ 34.55万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-15 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9177304
• 关键词: Address; Affect; Affinity; Alanine; Binding; Binding Proteins; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biology; Biophysics; Cell physiology; Cells; Chronic; Communication; Complex; Crystallography; DNA Sequence Alteration; Dependence; Development; Disease; Docking; Drug Targeting; Epitopes; Figs - dietary; Fluorescence; Fluorescence Resonance Energy Transfer; Genetic; Goals; Gold; Hot Spot; Human; Hyperactive behavior; I Kappa B-Alpha; Immune System Diseases; Immunologic Deficiency Syndromes; In Vitro; Inflammatory; Interdisciplinary Study; Kinetics; Knowledge; Lead; Length; Ligands; Location; Malignant Neoplasms; Maps; Measures; Mediating; Mediator of activation protein; Molecular; Molecular Conformation; Mutagenesis; Mutate; Mutation; Outcomes Research; Pathway interactions; Peptides; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Play; Principal Investigator; Process; Property; Proteins; Recombinants; Research; Resolution; Roentgen Rays; Role; Scaffolding Protein; Scanning; Shapes; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Site; Structural Models; Structure; Testing; Therapeutic Intervention; Ubiquitin; Work; X-Ray Crystallography; Zinc Fingers; assay development; base; dimer; disease-causing mutation; drug discovery; experience; functional outcomes; human disease; in vitro Assay; inhibitor/antagonist; insight; mutant; novel therapeutics; protein function; protein protein interaction; scaffold; small molecule; spatiotemporal; targeted treatment; therapeutic development

PROJECT SUMMARY/ABSTRACT Signal transduction pathways play pervasive and important roles in normal cellular physiology, and alterations in these pathways are found in many human diseases. One under-studied aspect of many signal transduction pathways is the role of scaffold proteins, which act as integrating platforms for signaling proteins. The NF-κB signaling pathway is altered in many inflammatory and immune diseases and cancers, and is thus viewed as target for therapeutic intervention. The overall project goal is to advance our understanding of the signaling scaffold protein NF-κB essential modulator (NEMO), a component of the inhibitor of κB kinase (IKK) complex, which is a key regulatory node for NF-κB signaling. In addition to NEMO playing a role in the chronic hyperactivity of NF-κB in human diseases, mutations in NEMO are found in several human immunodeficiency diseases. The long-term goals of the project are to understand how scaffolding proteins such as NEMO use conformational change to regulate the functional interactions between the signaling proteins that are bound to them, to elucidate the structural basis for disease-causing mutations in key regions of NEMO, and to identify new target sites for small molecule drugs that modulate NEMO activity. The following specific aims will be pursued: 1. Structural, biophysical and molecular approaches will be used to characterize the newly discovered “Intervening Domain” (IVD; aa 111-195) of NEMO, and to elucidate the role that the IVD plays in modulating IKKβ binding. Additionally, the mechanism(s) underlying immunodeficiency disease-causing mutations in the IVD region will be established. 2. The recently discovered interaction between NEMO and IκBα will be characterized using biochemical and cellular approaches, to establish whether this interface provides alternative target sites for therapeutic intervention. 3. The mechanism by which NEMO regulates the phosphorylation of IκB by IKKβ, and the role in this process played by a conformational change in NEMO, will be assessed and defined. The project team includes three Principal Investigators with expertise and experience across all aspects of the project, including assay development, quantitative and mechanistic biochemistry, X-ray crystallography and small-angle X-ray scattering (SAXS), biophysics, drug discovery, and NF-κB pathway biology and disease. The outcomes of this research will further our understanding of the role of NEMO in normal cellular physiology and in NEMO-related genetic immunodeficiencies, and may lead to the development of new therapeutics for human disease.

项目总结/摘要 信号转导通路在正常细胞生理学和改变中起着广泛而重要的作用。 在许多人类疾病中发现了这些途径。许多信号转导中一个未被充分研究的方面 在信号通路中，支架蛋白的作用是作为信号蛋白的整合平台。NF-κB 信号通路在许多炎症和免疫疾病和癌症中改变，因此被视为 治疗干预目标。整个项目的目标是推进我们对信号的理解 支架蛋白NF-κB必需调节剂（NEMO），κB激酶抑制剂（IKK）复合物的组分， 其是NF-κB信号传导的关键调节节点。除了NEMO在慢性疾病中发挥作用外， NF-κB在人类疾病中过度活性，NEMO的突变在几种人类免疫缺陷病毒中被发现。 疾病该项目的长期目标是了解NEMO等支架蛋白如何使用 构象变化，以调节信号蛋白之间的功能性相互作用，所述信号蛋白与 他们，阐明致病突变的结构基础，在关键区域的NEMO，并确定 用于调节NEMO活性的小分子药物的新靶点。以下具体目标将是 追求： 1.结构，生物物理和分子方法将用于表征新发现的 “干预领域”（IVD; aa 111-195），并阐明IVD在 调节IKKβ结合。此外，免疫缺陷疾病引起的潜在机制 将确定IVD区域中的突变。 2.最近发现的NEMO和IκBα之间的相互作用将使用生物化学和 细胞方法，以确定该界面是否为治疗提供了替代靶位点， 干预 3. NEMO调节IKKβ对IκB磷酸化的机制及其在此过程中的作用 由NEMO中的构象变化所起的作用，将被评估和定义。 项目团队包括三名主要研究人员，他们在项目的各个方面都有专业知识和经验。 项目，包括分析开发，定量和机械生物化学，X射线晶体学和 小角X射线散射（SAXS）、生物物理学、药物发现以及NF-κB通路生物学和疾病。的 这项研究的结果将进一步加深我们对NEMO在正常细胞生理学中的作用的理解， NEMO相关的遗传免疫缺陷，并可能导致开发新的治疗人类免疫缺陷的药物。 疾病