Structure-function studies on a key signaling module from interleukin 17 receptor

白细胞介素17受体关键信号模块的结构-功能研究

• 批准号: 8771995
• 负责人: Junpeng Deng
• 金额: $ 17.94万
• 依托单位: OKLAHOMA STATE UNIVERSITY STILLWATER
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8771995
• 关键词: Adaptor Signaling Protein; Adopted; Affinity; Autoimmune Diseases; Binding; Biological Assay; C-terminal; Cells; Complement Factor B; Gene Expression; IL17 gene; In Vitro; Inflammatory; Interleukin-17; Knowledge; Ligands; Maps; Mediating; Molecular; Mutagenesis; Nuclear; Peptides; Play; Proteins; Receptor Signaling; Recruitment Activity; Signal Transduction; Structure; Surface Plasmon Resonance; Tail; base; design; human disease; inhibitor/antagonist; novel strategies; novel therapeutics; protein aminoacid sequence; public health relevance; receptor; structural biology

DESCRIPTION (provided by applicant): Interleukin 17 (IL17) plays a key role in mediating inflammatory and autoimmune diseases. A unique intracellular signaling domain termed 'SEFIR' was identified within all IL17 receptors (IL17Rs) and the adaptor protein, nuclear factor ?B (NF-?B) activator 1 (Act1). A key step in IL17 signaling is the recruitment of Act1 to IL17Rs via SEFIR mediated associations. As the largest among all IL17Rs, IL17RA is a key receptor involved in essentially all IL17 signaling and requires a unique structural module (SEFEX) that is composed of both SEFIR and a short flanking C-terminal extension. The structure and mechanism of SEFEX-mediated IL17RA signaling are not known. In this proposal, we aim for providing molecular details of SEFEX domain from IL17RA, and revealing the molecular principles by which SEFEX recruits Act1 for IL17RA intracellular signaling. Aim 1. Structural and functional studies on the SEFEX domain of IL17RA. Hypothesis: The intracellular signaling module of IL17RA is unique among all IL17 receptors, and contains the SEFIR and a C-terminal flanking fragment of about 100 a.a. The IL17RA SEFIR domain itself adopts a unique structure, compared to that from other IL17Rs and Act1. I) We will determine the crystal structure of IL17RA-SEFEX. II) We will map the functional residues on IL17RA SEFEX that are critical for interaction with Act1 and IL17-stimulated signaling. We will use mutagenesis, CO-IP, surface plasmon resonance, and cell-based IL17 signaling assays. Aim 2. Determine the mechanism by which IL17RA recruits Act1 by structural and functional studies. Hypothesis: Helix ?C in the SEFIR domains is a conserved structural motif involved in heterotypic SEFIR- SEFIR interactions. Therefore peptides derived from the ?C helix of Act1, IL17RB or IL17RA will be inhibitory for IL17/IL25 signaling. I) We will determine the molecular mechanism by which IL17RA recruits Act1 by structural biology. II) We will develop cell-permeable inhibitory peptides for inhibition of IL17 signaling. These studies offer an opportunity for uncovering novel approaches for modulating IL17 and IL25 signaling. Our objectives are the detailed characterizations of the unique signaling domain from IL17RA, which is the most commonly shared receptor for nearly all IL17 ligands signaling. The expected results will provide detailed structural information for advancing our understanding on the fundamental principles of IL17 receptor signaling. Our studies will provide a platform for designing selective inhibitors that may be further developed into new therapeutics against a number of inflammatory and autoimmune diseases.

描述（由申请人提供）：白细胞介素17（IL17）在介导炎症和自身免疫性疾病中发挥关键作用。在所有 IL17 受体 (IL17R) 和接头蛋白、核因子 κB (NF-κB) 激活剂 1 (Act1) 中均发现了一个名为“SEFIR”的独特细胞内信号传导结构域。 IL17 信号转导的关键步骤是通过 SEFIR 介导的关联将 Act1 招募到 IL17R。作为所有 IL17R 中最大的受体，IL17RA 是基本上参与所有 IL17 信号传导的关键受体，并且需要一个由 SEFIR 和短侧翼 C 端延伸组成的独特结构模块 (SEFEX)。 SEFEX 介导的 IL17RA 信号传导的结构和机制尚不清楚。在本提案中，我们的目标是提供 IL17RA SEFEX 结构域的分子细节，并揭示 SEFEX 招募 Act1 进行 IL17RA 细胞内信号传导的分子原理。 目的 1. IL17RA SEFEX 结构域的结构和功能研究。假设：IL17RA 的细胞内信号传导模块在所有 IL17 受体中是独特的，并且包含 SEFIR 和约 100 个氨基酸的 C 端侧翼片段。与其他 IL17R 和 Act1 相比，IL17RA SEFIR 结构域本身采用独特的结构。 I) 我们将确定 IL17RA-SEFEX 的晶体结构。 II) 我们将绘制 IL17RA SEFEX 上的功能残基图谱，这些残基对于与 Act1 和 IL17 刺激的信号传导相互作用至关重要。我们将使用诱变、CO-IP、表面等离子共振和基于细胞的 IL17 信号转导测定。目标 2. 通过结构和功能研究确定 IL17RA 招募 Act1 的机制。假设：SEFIR 结构域中的 Helix ?C 是参与异型 SEFIR-SEFIR 相互作用的保守结构基序。因此，源自 Act1、IL17RB 或 IL17RA 的 ?C 螺旋的肽将抑制 IL17/IL25 信号传导。 I) 我们将通过结构生物学确定IL17RA招募Act1的分子机制。 II) 我们将开发细胞渗透性抑制肽来抑制 IL17 信号传导。这些研究为揭示调节 IL17 和 IL25 信号传导的新方法提供了机会。我们的目标是详细表征 IL17RA 的独特信号传导结构域，IL17RA 是几乎所有 IL17 配体信号传导最常见的受体。预期结果将为加深我们对 IL17 受体信号传导基本原理的理解提供详细的结构信息。我们的研究将为设计选择性抑制剂提供一个平台 进一步开发成针对多种炎症和自身免疫性疾病的新疗法。