IKK:Biophysical basis of dynamic regulation

IKK：动态调节的生物物理基础

• 批准号: 8072633
• 负责人: GOURISANKAR GHOSH
• 金额: $ 53.88万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8072633
• 关键词: Accounting; Adverse effects; Affect; Analytical Centrifugation; Architecture; Arthritis; Atherosclerosis; Binding; Binding Sites; Biochemical; Biochemical Reaction; Biochemistry; Biological Assay; Biophysics; Catalytic Domain; Cell Line; Cell Surface Receptors; Cells; Characteristics; Chronic; Chronic Disease; Complex; Computer Simulation; Coupled; Crystallography; Development; Disease; Dissociation; Dose; Drug Delivery Systems; Effectiveness; Enzymes; Equation; Estimation Techniques; Family; Feedback; Genes; Genetic; Goals; Half-Life; Heat-Shock Proteins 90; Human Pathology; IkappaB kinase; Immune; Immune response; In Vitro; Inflammatory; Inflammatory Response; Interleukin-1; Intervention; Kinetics; Lesion; Libraries; Malignant Epithelial Cell; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mediating; Methodology; Modeling; Molecular; Molecular Chaperones; Monitor; Multienzyme Complexes; NF-kappa B; Natural regeneration; Neoplasm Metastasis; Nucleotides; Outcome; Pathogenesis; Pathology; Pathway interactions; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacological Treatment; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Physiology; Play; Protein Dephosphorylation; Protein Kinase; Protein Phosphatase 2A Regulatory Subunit PR53; Proteins; Reaction; Receptor Cell; Receptor Down-Regulation; Recycling; Regulation; Reporting; Rest; Role; Scaffolding Protein; Signal Transduction; Signaling Protein; Specificity; Squamous cell carcinoma; Stimulus; Stream; Structure; Structure-Activity Relationship; Study models; TNF gene; Testing; Therapeutic; Therapeutic Intervention; Tumor Necrosis Factor Receptor; Work; autocrine; base; cancer cell; cancer type; cell type; crosslink; cytokine; design; high throughput screening; immune activation; in vivo; inhibitor/antagonist; mathematical model; mutant; novel; paracrine; public health relevance; reaction rate; receptor; reconstitution; research study; response; self assembly; simulation; small molecule; src-Family Kinases; text searching; transcription factor; ubiquitin ligase; upstream kinase

DESCRIPTION (provided by applicant): IKK is the major signaling hub for inflammatory and innate immune responses. It is an enzyme complex that receives signals from a large number of cellular receptors regulates that activity of the NF-kB family of transcription factors. Misregulation of IKK is associated with many chronic diseases, such as chronic inflammatory pathologies (arthritis, atherosclerosis, etc) and many different types of cancer. Genetic evidence strongly supports IKK's critical and central role in many functions in physiology and pathology, but its potential as a drug target has not been realized. Mechanistic and biophysical studies have been lacking, and we have neither a kinetic understanding of IKK regulation nor structural information. However, what is clear is that IKK mediates signaling specificity by tight dynamic control that is stimulus-specific and induces the expression of specific sets of genes. Recent studies have reported on a tightly coupled activation and inactivation mechanism that can only be described by a multi-state activation-inactivation cycle that involves the function of several enzymes with different functions, such as ubiquitin ligases, kinases, phosphatases, and foldases. Through regulation of these activities, we hypothesize that the IKK cycle is driven in a stimulus- and cell type-specific manner, and that understanding the kinetic relationships will reveal opportunities for rationally targeted pharmacological intervention that discriminate between disease associated misregulation and stimulus-responsive regulation in healthy cells. In this proposal, we will construct a mathematical model of the IKK cycle to explore the dynamic regulation of IKK activity. We will then focus biochemical and biophysical studies on specific control mechanisms. In particular, we test the roles of IKK oligomerization, conformational changes, and upstream kinases in IKK activation and inhibition of IKK. Computational simulations will guide genetic and pharmacological manipulation of IKK dynamics. Finally, we will focus our study on how TNF and IL-1 produce differential dynamic control of IKK; how differential dose response and temporal control determine the efficacy of cytokine traps. PUBLIC HEALTH RELEVANCE: The IkappaB kinase (IKK) complex is large protein kinase that specifically receives signals from cell surface receptors and transmits the signal to downstream effector transcription factor NF-kB. IKK must remain at a low activity state under resting state of the cell and transits into a high activity state upon receiving signals. Any alteration of this tight regulatory mode of IKK results to abnormal cellular outcomes including inflammatory diseases and cancer. The precise mechanism of how IKK-intrinsic and extrinsic cellular mechanisms control IKK activity is poorly understood. This proposal will integrate diverse methodologies that include mathematical modeling, biophysics, biochemistry and genetics to probe IKK regulation.

描述（由申请人提供）：IKK是炎症和先天免疫反应的主要信号中心。这是一种酶复合物，从大量细胞受体中接收信号可调节NF-KB转录因子家族的活性。 IKK的不调节与许多慢性疾病有关，例如慢性炎症病理学（关节炎，动脉粥样硬化等）和许多不同类型的癌症。遗传证据强烈支持IKK在生理和病理学中许多功能中的关键和核心作用，但尚未实现其作为药物靶标的潜力。机械和生物物理研究一直缺乏，我们对IKK调节也没有动力学的理解，也没有结构信息。但是，很明显，IKK通过刺激特异性的紧密动态控制介导信号传导特异性，并诱导特定基因集的表达。最近的研究报道了一种紧密耦合的激活和灭活机制，该机制只能通过多态激活灭活周期来描述，该循环涉及几种具有不同功能的酶的功能，例如泛素连接酶，激酶，磷酸酶和折叠酶。通过调节这些活动，我们假设IKK周期是以刺激和细胞类型特异性的方式驱动的，并且理解动力学关系将揭示有理由靶向的药理学干预的机会，以区分疾病相关的错误调节和健康细胞的刺激反应性调节。在此建议中，我们将构建IKK循环的数学模型，以探索IKK活动的动态调节。然后，我们将将生化和生物物理研究集中在特定的控制机制上。特别是，我们测试了IKK寡聚，构象变化和上游激酶在IKK激活和抑制IKK中的作用。计算模拟将指导IKK动力学的遗传和药理学操纵。最后，我们将重点关注TNF和IL-1如何产生IKK的差异动态控制。差异剂量反应和时间控制如何确定细胞因子陷阱的功效。 公共卫生相关性：Ikappab激酶（IKK）复合物是大蛋白激酶，它专门从细胞表面受体接收信号，并将信号传输到下游效应器转录因子NF-KB。在接收信号后，IKK必须保持在细胞静止状态下的低活动状态，并在高活动状态下转变为高活动状态。这种紧密的IKK调节模式的任何改变会导致细胞异常结果，包括炎症性疾病和癌症。 IKK intrinsic和外部细胞机制如何控制IKK活性的确切机制知之甚少。该建议将整合包括数学建模，生物物理学，生物化学和遗传学的多种方法，以探测IKK调节。