IKK:Biophysical basis of dynamic regulation

IKK：动态调节的生物物理基础

基本信息

批准号：
7714251
负责人：
GOURISANKAR GHOSH
金额：
$ 54.46万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/7714251
关键词：
Accounting Adverse effects Affect Analytical Centrifugation Architecture Arthritis Atherosclerosis Binding 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 Metastatic Squamous Cell Carcinoma 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 Stimulus Stream Structure Structure-Activity Relationship 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 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

项目摘要

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传导中心。它是一种酶复合物，接收来自大量细胞受体的信号，调节 NF-kB 转录因子家族的活性。 IKK 的失调与许多慢性疾病有关，例如慢性炎症病理（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 在生理学和病理学许多功能中的关键和核心作用，但其作为药物靶点的潜力尚未实现。一直缺乏机制和生物物理学研究，我们既没有对 IKK 调控的动力学理解，也没有结构信息。然而，显而易见的是，IKK 通过刺激特异性的严格动态控制来介导信号特异性，并诱导特定基因组的表达。最近的研究报告了紧密耦合的激活和失活机制，该机制只能通过多状态激活-失活循环来描述，该循环涉及多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。通过对这些活动的调节，我们假设 IKK 循环以刺激和细胞类型特异性的方式驱动，并且了解动力学关系将揭示合理有针对性的药物干预的机会，从而区分疾病相关的失调和刺激响应调节在健康细胞中。在本提案中，我们将构建IKK循环的数学模型，以探索IKK活动的动态调控。然后我们将重点研究特定控制机制的生化和生物物理研究。特别是，我们测试了 IKK 寡聚化、构象变化和上游激酶在 IKK 激活和抑制中的作用。计算模拟将指导 IKK 动力学的遗传和药理学操作。最后，我们将重点研究TNF和IL-1如何对IKK产生差异动态控制；不同的剂量反应和时间控制如何决定细胞因子陷阱的功效。公共健康相关性：IkappaB 激酶 (IKK) 复合物是一种大型蛋白激酶，专门接收来自细胞表面受体的信号并将信号传输至下游效应转录因子 NF-kB。 IKK在细胞静息状态下必须保持在低活性状态，并在接收到信号后转变为高活性状态。 IKK 这种严格调节模式的任何改变都会导致异常的细胞结果，包括炎症性疾病和癌症。 IKK 内在和外在细胞机制如何控制 IKK 活性的精确机制尚不清楚。该提案将整合数学模型、生物物理学、生物化学和遗传学等多种方法来探索 IKK 调控。