Structural biology of shared cytokine signaling receptors

共享细胞因子信号受体的结构生物学

• 批准号: 7644303
• 负责人: Kenan Christopher GARCIA
• 金额: $ 29.25万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7644303
• 关键词: Address; Architecture; Autoimmune Diseases; Autoimmunity; Award; Binding; Biochemistry; Biological Assay; Calorimetry; Cell membrane; Cell surface; Chemistry; Ciliary Neurotrophic Factor; Communication; Complement component C5; Complex; Coupling; Crystallization; Crystallography; Cytokine Receptors; Cytokine Signaling; Electron Microscopy; Electrons; Event; Extracellular Structure; Family; Foundations; Functional disorder; Glycoproteins; Goals; Growth Factor; Hematopoietic; Homeostasis; IL27RA gene; Image; Immune; Interleukin 2 Receptor Gamma; Interleukin-11; Interleukin-13; Interleukin-15; Interleukin-2; Interleukin-4; Interleukin-6; Investigation; Knowledge; LIF gene; Length; Ligands; Malignant Neoplasms; Measures; Mediating; Membrane Proteins; Microscopic; Molecular; Nervous system structure; Physiological Processes; Protein Binding; Proteins; Receptor Signaling; STAT protein; Shapes; Signal Transduction; Solutions; Structure; Structure-Activity Relationship; System; Techniques; Therapeutic; Titrations; base; cell growth; cross reactivity; cytokine; cytokine receptor gp130; design; extracellular; human disease; immune function; insight; interest; interleukin-13 receptor; interleukin-15 receptor; leukemia inhibitory factor; leukemia inhibitory factor receptor; receptor; receptor binding; reconstitution; research study; structural biology

DESCRIPTION (provided by applicant): Type-I four-helix bundle cytokines mediate a wide range of physiological processes through their effects on cell growth, differentiation, and proliferation. Collectively, through both pleiotropic and unique activities, they contribute to the homeostasis of the immune, hematopoietic, and nervous systems. The majority of hematopoietic and immune regulatory cytokines act through a small group of shared transmembrane signaling receptors. The two most widely used shared receptors are gp130, and the common gamma chain (3c), which engage their cytokine ligands through their extracellular regions, resulting in activation of intracellular JAK/STAT signaling cascades. Gp130 is the shared receptor for eleven cytokines (IL-6, IL-11, Ciliary Neurotrophic Factor, Oncostatin, Leukemia Inhibitory Factor, and others), and 3c is the shared receptor for 6 cytokines (IL-2, -4, -7,- 9, -15, -21). During the prior term of this award, we elucidated structural architectures of the two paradigmatic receptor complexes in each family, gp130/IL-6 and 3c/IL-2. These ternary and quaternary complexes represent blueprints for the assembly of receptor-ligand complexes used by all ligands in each shared receptor family. In this proposal, we escalate our investigations into the structural biology of these shared receptors in regards to extracellular ligand recognition, assembly of hetero-oligomeric cell surface complexes, transmembrane signaling, and intracellular activation of JAK and STAT. In order to do this, we are: 1- determining x-ray crystal structures of soluble extracellular multimeric receptor-ligand complexes, 2- measuring the cooperativity and assembly energetics of stepwise cytokine-receptor complex formation, 3- purifying the entire full-length receptor complexes, as membrane proteins, bound to both cytokine and intracellular adaptors and imaging them by electron microscopy, 4- reconstituting the intracellular signaling complexes consisting of JAK and STAT, and 5- complementing the crystallographic studies with NMR to interrogate potential allostery during signaling. In this fashion, by taking a multi-disciplinary strategy we propose to obtain a complete molecular snapshot of a shared signaling receptor from the initial engagement of ligand through the activation of intracellular signaling cascades. PROJECT NARRATIVE: Growth factors termed cytokines, and their receptors, are essential to the normal functioning of the immune, hematopoietic, and nervous systems, and dysfunction of the growth factor network underlies numerous human disease conditions such as autoimmunity and cancer. We propose to use the techniques of biochemistry and structural biology to visualize the three-dimensional shapes of growth factors bound to their cellular receptors, and understand how this binding event is communicated across the cell membrane. These studies will give us insight into basic receptor signaling mechanisms, as well as facilitate the manipulation of the cytokine receptor system for therapeutic purposes.

描述(由申请人提供)：I型四股螺旋束细胞因子通过对细胞生长、分化和增殖的影响，调节广泛的生理过程。总体而言，通过多效性和独特的活性，它们有助于免疫、造血和神经系统的动态平衡。大多数造血和免疫调节细胞因子通过一小群共享的跨膜信号受体发挥作用。两个使用最广泛的共享受体是gp130和共同的伽马链(3c)，它们通过细胞外区与细胞因子配体结合，导致细胞内JAK/STAT信号级联激活。Gp130是11种细胞因子(IL-6、IL-11、睫状神经营养因子、抑瘤素、白血病抑制因子等)的共同受体，3c是6种细胞因子(IL-2、-4、-7、-9、-15、-21)的共同受体。在本奖项的前一期，我们阐明了每个家族中两个聚集性受体复合体的结构，gp130/IL-6和3c/IL-2。这些三元和四元络合物代表了每个共享受体家族中所有配体使用的受体-配体络合物的组装蓝图。在这个提案中，我们升级了对这些共享受体的结构生物学的研究，涉及细胞外配体识别、异寡聚细胞表面复合体的组装、跨膜信号转导以及JAK和STAT的细胞内激活。为此，我们进行了以下工作：1-测定可溶胞外多聚体受体-配体复合体的X射线晶体结构，2-测量逐步形成的细胞因子-受体复合体的协作性和组装能量学，3-纯化整个全长受体复合体作为膜蛋白，与细胞因子和细胞内接头结合并通过电子显微镜成像，4-重建由JAK和STAT组成的细胞内信号复合体，以及5-用核磁共振补充结晶学研究以询问信号传递过程中的潜在变构。以这种方式，通过采取多学科策略，我们建议通过激活细胞内信号级联，从配体的初始参与获得共享信号受体的完整分子快照。项目简介：被称为细胞因子及其受体的生长因子对免疫、造血和神经系统的正常功能是必不可少的，生长因子网络的功能障碍是许多人类疾病(如自身免疫和癌症)的基础。我们建议使用生物化学和结构生物学技术来可视化生长因子与其细胞受体结合的三维形状，并了解这种结合事件是如何通过细胞膜传递的。这些研究将使我们深入了解基本的受体信号机制，以及促进细胞因子受体系统的治疗目的的操纵。