SAXS OF THE COMPLEX OF ANTHRAX TOXINS AND HUMAN INSULIN DEGRADING ENZYME

炭疽毒素与人胰岛素降解酶复合物的SAXS

• 批准号: 8168652
• 负责人: WEI-JEN TANG
• 金额: $ 0.36万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168652
• 关键词: Address; Adenylate Cyclase; Amyloid beta-Protein; Anthrax disease; Bacillus anthracis; Bacteria; Bacterial Toxins; Binding; Binding Sites; Biological Process; Bordetella pertussis; Calmodulin; Cell Surface Receptors; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Diabetes Mellitus; Disease; Enzymes; Funding; Grant; Human; Institution; Insulin; Insulinase; Lipase; Metalloproteases; Mitochondria; Peptide Hydrolases; Pertussis; Play; Process; Proteins; Research; Research Personnel; Resources; Role; Set protein; Solutions; Source; Structure; Toxin; United States National Institutes of Health; anthrax edema factor; anthrax protective factor; anthrax toxin; edema factor; insight; protein protein interaction

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Protein-protein interaction is a process that controls the diverse biological functions. We will use SAXS to address the complex protein-protein interactions in solution by studying human proteases and bacterial toxins. First, we will study two human metalloproteases, insulin degrading enzyme (IDE) and presequence peptidase (PreP). IDE plays a key role in the clearance of insulin and amyloid beta, peptides vital for the progression of diabetes and Alzheimer¿s disease, respectively. PreP degrades mitochondrial presequences and amyloid beta so that it plays a key role to maintain integrity of mitochondria. We propose to use SAXS on study the oligomerization, the conformational switch, and substrate binding of these enzymes. Second set of proteins are bacterial toxins secreted by Bacillus anthracis, and Bordetella pertussis, the etiologic agent for anthrax and whooping cough. Our lab has solved the crystal structures of anthrax edema factor (EF), anthrolysin O (ALO), and pertussis CyaA. EF is an adenylyl cyclase toxin that binds anthrax protective antigen for its cell entrance and binds calmodulin for its catalytic activation. Pertussis CyaA also requires the binding of calmodulin for its catalytic activation. However CyaA has the calmodulin-binding site distinct from that of EF. For ALO, it needs to oligomerize for its pore-forming activity as well as binds to cell surface receptor. In addition, anthrax bacteria secrete approximately 400 proteins and of those, we have chosen several lipases and proteases to study their functions. SAXS will be used to gain the new insights for the interaction of these toxins with their partner proteins.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 蛋白质-蛋白质相互作用是控制多种生物功能的过程。我们将使用 SAXS 通过研究人类蛋白酶和细菌毒素来解决溶液中复杂的蛋白质-蛋白质相互作用。首先，我们将研究两种人类金属蛋白酶：胰岛素降解酶（IDE）和前序列肽酶（PreP）。 IDE 在胰岛素和β淀粉样蛋白的清除中发挥着关键作用，这两种肽分别对糖尿病和阿尔茨海默病的进展至关重要。 PreP 会降解线粒体前序列和 β 淀粉样蛋白，因此在维持线粒体完整性方面发挥着关键作用。我们建议使用 SAXS 来研究这些酶的寡聚、构象转换和底物结合。第二组蛋白质是炭疽杆菌和百日咳博德特氏菌（炭疽和百日咳的病原体）分泌的细菌毒素。我们实验室已经解析了炭疽水肿因子（EF）、炭疽溶血素O（ALO）和百日咳CyaA的晶体结构。 EF 是一种腺苷酸环化酶毒素，可结合炭疽保护性抗原进入细胞，并结合钙调蛋白催化激活。百日咳 CyaA 还需要结合钙调蛋白来催化激活。然而，CyaA 的钙调蛋白结合位点与 EF 不同。对于 ALO，它需要寡聚化以发挥其成孔活性并与细胞表面受体结合。此外，炭疽菌分泌大约 400 种蛋白质，其中，我们选择了几种脂肪酶和蛋白酶来研究它们的功能。 SAXS 将用于获得这些毒素与其伙伴蛋白相互作用的新见解。