Molecular Recognition Studies of the Interleukin-7 Pathway

IL-7 通路的分子识别研究

• 批准号: 8080545
• 负责人: SCOTT T WALSH
• 金额: $ 21.4万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-15 至 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8080545
• 关键词: Affinity; Agonist; Allergic; Asparagine; Autoimmune Process; B-Lymphocytes; Binding; Biological; Biological Assay; Calorimetry; Cardiovascular Diseases; Cell Energetics; Cell Proliferation; Cell Surface Receptors; Cells; Complex; Coronary Arteriosclerosis; Crystallography; Deficiency Diseases; Development; Disease; Engineering; Epitopes; Event; Extracellular Domain; Extracellular Matrix; Family; Generations; Genetic Transcription; Goals; Heterodimerization; Homeostasis; Human; IL7R gene; Interleukin 2 Receptor Gamma; Interleukin 7 Receptor; Interleukin-7; Kinetics; Knowledge; Length; Link; Malignant Neoplasms; Membrane; Methods; Modeling; Molecular; Molecular Conformation; Mutagenesis; Mutation; NMR Spectroscopy; Pathogenesis; Pathway interactions; Peptides; Phage Display; Play; Positioning Attribute; Property; Proteins; Relative (related person); Research; Research Proposals; Resolution; Role; Rotation; Scanning; Severe Combined Immunodeficiency; Signal Pathway; Signal Transduction; Site; Specificity; Structure; Surface Plasmon Resonance; Techniques; Thermodynamics; Titrations; Variant; cytokine; design; directed evolution; disease phenotype; driving force; engineering design; glycosylation; human TSLP protein; leukemia/lymphoma; molecular recognition; novel; novel strategies; novel therapeutics; public health relevance; receptor; response; three dimensional structure

DESCRIPTION (provided by applicant): Interleukin-7 (IL-7) is an essential pleiotrophic cytokine regulating numerous immunological events. Normal IL-7 signaling responses trigger the development, proliferation, and homeostasis of T and B cells as well as remodeling of the extracellular matrix. IL-7 activates its signaling cascade by interacting with its own cell surface receptor, IL-7R alpha, and the common gamma chain receptor through a cytokine-induced receptor heterodimerization mechanism. Under- and over-stimulation of the IL-7 pathway has been implicated in the pathogenesis of a form of severe combined immunodeficiency, autoimmune conditions, coronary artery disease, and several cancers. Structural, energetic, and cell biological studies of IL-7-induced receptor heterodimerization require a careful analysis of two distinct events: the association of IL-7 to IL-7R alpha and of IL-7:IL-7R alpha complex to the common gamma chain receptor. This research proposal will focus on the initiation step, the binding interaction of IL-7 with its alpha receptor. Results show that glycosylation of the IL- 7R alpha confers optimal binding affinity to IL-7. The importance of glycosylation of the IL-7R alpha appears to be a novel binding recognition mechanism for the common gamma chain receptor family. The thermodynamic and kinetic properties of the wild-type IL-7:IL-7R alpha interaction will be ascertained using a variety of biophysical methods including isothermal titration calorimetry and surface plasmon resonance. The functional epitopes on both IL-7 and IL-7R alpha will be determined using site-directed scanning mutagenesis, biophysical methods, and cell proliferation assays. Three-dimensional structures of free IL-7, free unglycosylated and glycosylated IL-7R alpha, and complexes of IL-7 bound to unglycosylated and glycosylated IL-7R alpha will be determined using x-ray crystallography or NMR spectroscopy. Our understanding of the binding determinants for the IL-7-receptor interaction will be used to engineer novel peptide and protein antagonists using rational protein design and phage display techniques. The long-term goal of this research involves development of a structural energetic model to explain the normal and aberrant IL-7 signaling mechanisms and the generation of new therapeutic peptides/proteins to treat the many deficiencies and diseases associated with impaired IL-7 signaling. Public Health Relevance Statement: This research proposal will focus on the structural, energetic, and cell biological studies of the interaction between human interleukin-7 and its receptor, interleukin-7 receptor alpha. Our understanding of the binding determinants for the IL-7-receptor interaction will be used to engineer novel peptide and protein antagonists using rational protein design and phage display techniques. The long-term goal of this research involves development of a structural energetic model to explain the normal and aberrant IL-7 signaling mechanisms and the generation of new therapeutic peptides/proteins to treat the many deficiencies and diseases associated with impaired human IL-7 signaling.

描述（由申请人提供）：白细胞介素-7 （IL-7）是一种重要的多营养细胞因子，可调节许多免疫事件。正常的IL-7信号反应触发T细胞和B细胞的发育、增殖和稳态，以及细胞外基质的重塑。IL-7通过细胞因子诱导的受体异二聚化机制，与自身细胞表面受体IL-7R α和常见γ链受体相互作用，激活其信号级联。IL-7通路的不足和过度刺激与一种严重的联合免疫缺陷、自身免疫性疾病、冠状动脉疾病和几种癌症的发病机制有关。IL-7诱导受体异二聚化的结构、能量和细胞生物学研究需要仔细分析两个不同的事件：IL-7与IL-7R α的关联以及IL-7:IL-7R α复合物与常见γ链受体的关联。本研究计划将重点关注启动步骤，即IL-7与其α受体的结合相互作用。结果表明，IL- 7R α的糖基化使其与IL-7具有最佳的结合亲和力。IL-7R α糖基化的重要性似乎是常见γ链受体家族的一种新的结合识别机制。野生型IL-7:IL-7R α相互作用的热力学和动力学性质将通过各种生物物理方法确定，包括等温滴定量热法和表面等离子体共振。IL-7和IL-7R α的功能表位将通过定点扫描诱变、生物物理方法和细胞增殖试验来确定。游离IL-7、游离未糖基化和糖基化IL-7R α以及IL-7与未糖基化和糖基化IL-7R α结合的复合物的三维结构将使用x射线晶体学或核磁共振波谱学来确定。我们对il -7受体相互作用的结合决定因素的理解将用于利用合理的蛋白质设计和噬菌体展示技术来设计新的肽和蛋白质拮抗剂。本研究的长期目标包括建立一个结构能量模型来解释正常和异常的IL-7信号传导机制，并产生新的治疗肽/蛋白来治疗与IL-7信号传导受损相关的许多缺陷和疾病。