ANALYZE THE COMPLEX PROTEIN ASSEMBLY USING SAXS

使用 SAXS 分析复杂的蛋白质组装

• 批准号: 8361305
• 负责人: WEI-JEN TANG
• 金额: $ 0.59万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361305
• 关键词: Allosteric Regulation; Alzheimer' s Disease; Amyloid; Anthrax disease; Antigens; Area; Bacteria; Binding; Biological Process; Biophysics; Cell Wall; Complex; Diabetes Mellitus; Enzymes; Funding; Grant; Human; Insulin; Insulinase; Macrophage Inflammatory Protein-1; Macrophage Inflammatory Proteins; Metalloproteases; Methods; Molecular; Molecular Weight; National Center for Research Resources; Pathologic; Peptide Hydrolases; Peptides; Polymers; Principal Investigator; Property; Research; Research Infrastructure; Resources; Shapes; Signal Transduction; Solutions; Source; Structure; Toxin; United States National Institutes of Health; Virulence Factors; Work; anthrax lethal factor; anthrax toxin; base; cost; edema factor; novel; protein complex; protein protein interaction; response; retinal rods

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Our lab focuses on the molecular basis of protein-protein interaction crucial for cell signal transduction in the normal and pathologic responses of humans. Currently, we work on two major areas. First is the interaction of human protease, insulin-degrading enzyme (IDE) with its substrates. IDE, a 110 kDa metalloprotease, is involved in the clearance of insulin and amyloid ¿, peptides crucial for the progression of diabetes and Alzheimers' disease. We have used SAXS to probe the open-closed conformational switch regulated by substrate binding as well as the allosteric regulation by the oligomerization of IDE. We recently identify macrophage inflammatory protein (MIP)-1¿ and MIP-1¿ as the novel substrate of IDE. MIP-1¿ and MIP-1¿ form high molecular weight aggregates. We used SAXS to demonstrate that MIP-1¿ and MIP-1¿ form the rod-shaped polydisperse polymers in solution. I propose to continue to use SAXS to understand the structure features of IDE and its substrates and how they interact to achieve their biological functions. Second is the interactions of toxins secreted by bacteria that cause anthrax. Anthrax bacteria also secrete >200 secreted factors. This includes three major anthrax toxins, protective antigen (PA), edema factor (EF), and lethal factor (LF) and the protein complex of PA-EF and PA-LF. We use SAXS analysis as a method to probe the structural features of purified anthrax toxins and toxin complex. We have also chosen 10 of such factors including pore-forming toxins, proteases, and cell wall modifying enzymes and will apply SAXS to probe the biophysical properties of these virulence factors.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 我们的实验室专注于蛋白质-蛋白质相互作用的分子基础，这对于人类正常和病理反应中的细胞信号转导至关重要。目前，我们在两个主要领域开展工作。首先是人类蛋白酶、胰岛素降解酶（IDE）与其底物的相互作用。 IDE 是一种 110 kDa 金属蛋白酶，参与胰岛素和淀粉样蛋白的清除，这些肽对于糖尿病和阿尔茨海默病的进展至关重要。我们使用 SAXS 来探测底物结合调节的开闭构象开关以及 IDE 寡聚化的变构调节。我们最近确定巨噬细胞炎症蛋白 (MIP)-1¿ 和 MIP-1¿ 作为 IDE 的新型底物。 MIP-1¿和MIP-1¿形成高分子量聚集体。我们使用 SAXS 证明 MIP-1¿ 和 MIP-1¿ 在溶液中形成棒状多分散聚合物。我建议继续使用SAXS来了解IDE及其底物的结构特征以及它们如何相互作用以实现其生物学功能。其次是引起炭疽的细菌分泌的毒素的相互作用。炭疽细菌还分泌超过 200 种分泌因子。这包括三种主要的炭疽毒素：保护性抗原（PA）、水肿因子（EF）和致死因子（LF）以及PA-EF和PA-LF的蛋白质复合物。我们使用 SAXS 分析作为探测纯化炭疽毒素和毒素复合物结构特征的方法。我们还选择了其中的 10 个因子，包括成孔毒素、蛋白酶和细胞壁修饰酶，并将应用 SAXS 来探测这些毒力因子的生物物理特性。