Structural and functional studies of the IkappaB kinase (IKK) complex

IkappaB 激酶 (IKK) 复合物的结构和功能研究

• 批准号: 8278697
• 负责人: Hao Wu
• 金额: $ 3.55万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-25 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8278697
• 关键词: Attention; Baculoviruses; Binding; Biochemical; Biological; C-terminal; Calorimetry; Cell Nucleus; Cells; Complex; Crystallization; Data; Disease; Electron Microscopy; Enzyme Kinetics; Family; Genetic Transcription; HTATIP2 gene; Heart; Helix-Loop-Helix Motifs; Hereditary Disease; Human; Human Herpesvirus 8; IkappaB kinase; Immune response; Inflammatory; Inflammatory Response; Insecta; Interleukin-1 Receptors; Length; Leucine Zippers; Ligation; Link; Malignant Neoplasms; Mediating; Molecular; Molecular Conformation; Mutation; N-terminal; Nuclear; Oncogene Proteins; Paper; Pathway interactions; Phosphorylation; Phosphotransferases; Polyubiquitin; Polyubiquitination; Proteins; Publishing; Receptors, Antigen, B-Cell; Reporting; Series; Signal Transduction; Structure; Surface Plasmon Resonance; System; T-Cell Receptor; Titrations; Toll-like receptors; Tumor Necrosis Factor Receptor; Tumor Necrosis Factor-alpha; Ubiquitin; Virus Diseases; Zinc Fingers; base; cytokine; design; dimer; genetic regulatory protein; human disease; image reconstruction; inhibitor/antagonist; kinase inhibitor; light scattering; member; microbial; molecular mass; multicatalytic endopeptidase complex; mutant; protein protein interaction; receptor; response; therapeutic target; transcription factor

ABSTRACT Transcription factors in the nuclear factor ¿B (NF-¿B) family are evolutionarily conserved master regulators of immune and inflammatory responses. They are activated in response to ligation of many receptors including T-cell receptors, B-cell receptors, members of the tumor necrosis factor (TNF) receptor superfamily and the Toll-like receptor/interleukin-1 receptor (TLR/IL-1R) superfamily. The I¿B kinase (IKK), comprising IKK¿ and IKK¿, is at the heart of NF-¿B activation and mediates two NF-¿B activation pathways. The canonical NF-¿B pathway is triggered by microbial and viral infections and pro-inflammatory cytokines and is dependent on IKK¿ phosphorylation and activation. The alternative pathway is triggered by certain members of the TNF cytokine family and selectively activates IKK¿. Activated IKK phosphorylates I¿Bs, leading to their polyubiquitination and subsequent degradation by the proteasome. The freed NF-¿B dimers translocate to the nucleus to mediate transcription. Because of its importance in NF-¿B activation, IKK, especially IKK¿, has become a potential therapeutic target for many human diseases. The regulatory protein NEMO (also known as IKK¿ or FIP-3) interacts with IKK¿ and/or IKK¿ to form the IKK¿, IKK¿ or IKK¿/¿ holo-complex. The intact IKK¿ holo-complex is approximately 700-900kD in molecular mass containing multiple copies of IKK¿ and NEMO. IKK¿ and IKK¿ both contain the following conserved recognizable domains: a kinase domain (KD), a leucine zipper domain (LZ), a helix loop helix domain (HLH) and a C-terminal NEMO-binding domain (NBD). NEMO contains an N-terminal kinase-binding domain (KBD), a minimal oligomerization domain (MOD) that is also the ubiquitin binding domain (UBD) and a C-terminal zinc finger domain (ZF). IKK and NF-¿B signaling has attracted tremendous attention with more than 30,000 papers published on the subject. Despite the biological importance, not a single successful structure determination has been reported on IKK, an indication on the difficulty of the project. To elucidate the molecular basis of IKK function and to assist the discovery of IKK inhibitors, we propose a series of structural and functional studies on IKK, in particular, IKK¿ and its regulatory protein NEMO.

抽象的 核因子 ¿B (NF-¿B) 家族中的转录因子是进化上保守的主宰 免疫和炎症反应的调节剂。它们响应于连接而被激活 许多受体包括T细胞受体、B细胞受体、肿瘤坏死成员 因子 (TNF) 受体超家族和 Toll 样受体/白细胞介素 1 受体 (TLR/IL-1R) 超家族。 IKB 激酶（IKK）由 IKKK 和 IKKK 组成，是 NF-KB 激活的核心， 介导两个 NF-¿B 激活途径。典型的 NF-¿B 通路是由 微生物和病毒感染以及促炎细胞因子，并且依赖于 IKK¿ 磷酸化和激活。替代途径是由某些成员触发的 TNF 细胞因子家族并选择性激活 IKK¿。活化的 IKK 磷酸化 I¿B，导致 它们的多泛素化和随后被蛋白酶体降解。释放的 NF-¿B 二聚体易位至细胞核以介导转录。由于其在 NF-¿B 中的重要性 激活，IKK，特别是 IKK¿，已成为许多人类的潜在治疗靶点 疾病。 调节蛋白 NEMO（也称为 IKK¿ 或 FIP-3）与 IKK¿ 和/或 IKK¿ 相互作用， 形成 IKK¿、IKK¿ 或 IKK¿/¿ 全息复合体。完整的 IKK¿ 全息复合体大约是 分子量为 700-900kD，含有 IKK¿ 和 NEMO 的多个拷贝。 IKK¿ 和 IKK¿ 两者都包含以下保守的可识别结构域：激酶结构域 (KD)、亮氨酸 拉链结构域 (LZ)、螺旋环螺旋结构域 (HLH) 和 C 端 NEMO 结合结构域 （NBD）。 NEMO 包含一个 N 端激酶结合域 (KBD)，一个最小的寡聚体 结构域 (MOD)，也是泛素结合结构域 (UBD) 和 C 端锌指 域（ZF）。 IKK 和 NF-¿B 信号传导引起了极大关注，论文发表超过 30,000 篇 发表了关于该主题的文章。尽管具有生物学重要性，但没有一个成功的结构 IKK 已报告了该决定，这表明了该项目的难度。到 为了阐明 IKK 功能的分子基础并协助发现 IKK 抑制剂，我们 提出了一系列关于 IKK 的结构和功能研究，特别是 IKK¿ 及其调控 蛋白质尼莫。