STRUCTURAL STUDIES OF YEAST RIBONUCLEOTIDE REDUCTASE, AMYLOID-RECOGNIZING ANT

酵母核糖核苷酸还原酶、淀粉样蛋白识别蚂蚁的结构研究

• 批准号: 7956850
• 负责人: Chris G Dealwis
• 金额: $ 2.83万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7956850
• 关键词: Address; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Antineoplastic Agents; Antiviral Agents; Ants; Binding; Biochemical; C-terminal; Catalytic Domain; Clofarabine; Complex; Computer Retrieval of Information on Scientific Projects Database; Consensus; DNA biosynthesis; Data; Docking; Drug Design; Funding; Grant; Institution; Libraries; Modeling; Molecular; Mus; Pathogenesis; Peptides; Reporting; Research; Research Personnel; Resources; Ribonucleotide Reductase; Site; Source; Structure; Structure-Activity Relationship; United States National Institutes of Health; Yeasts; base; design; inhibitor/antagonist; mutant; neutralizing antibody; peptidomimetics; structural biology

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. Ribonucleotide reductase (RNR) catalyzes the rate-limiting step of de novo DNA synthesis by reducing NDPs to dNDPs. Though much structural and biochemical data on non-catalytic subunits of eukaryotic RNR exist, specifically mouse Rnr2, and yeast Rnr2p?Rnr4p, until now there is no reported structure for any eukaryotic Rnr1. Using MAD data from BioCARS, we solved the yeast Rnr1 structure. During this proposal we will solve several cognate effector-substrate complex structures to address how substrates are selected by specific dNTPs binding at the effector site. The yeast RNR assembly involves association of Rnr2-Rnr4 with Rnr1. C-terminal peptides of Rnr2 and Rnr4 can disrupt this assembly, and a new class of anticancer and antiviral inhibitors is designed based on Rnr2 peptides. We will solve structures of Rnr1 complexed with peptidomimetic libraries and anticancer agents like clofarabine. In yeast RNR activity is controlled allosterically by ATP (upregulator) and dATP (downregulator). We will solve the structures of Rnr1 complexed with ATP and dATP to address this. Yeast RNR is also downregulated by Sml1, which binds Rnr1. The C-termini of Sml1 inhibits RNR activity with reduced potency. We will solve the Sml1 structure using MAD and Rnr1 complexed with Sml1-derived peptides and intact Sml1. We have constructed numerous mutants of Rnr1to study structure-function relationships. Several have been crystallized both in the native form and in complex with effector-substrate complexes. Though accepted that ABeta is involved in the pathogenesis of Alzheimer?s, there is no consensus on its atomic structure. We will obtain this using neutralizing antibodies (Abs). The structures of the Abs alone can be used with models of AB for docking studies, or better, AB?Ab complexes can provide a molecular basis for designing drugs against Alzheimer?s. We have crystallized an AB-recognizing Fab in apo form and complexed to a truncated A¿ peptide, and several other AB-recognizing Abs.

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。 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以出现在其他 CRISP 条目中 列出的机构是。 对于中心来说，它不一定是研究者的机构。 核糖核苷酸还原酶 (RNR) 通过将 NDP 还原为 dNDP 来催化 DNA 从头合成的限速步骤，尽管迄今为止存在大量关于真核 RNR 非催化亚基的结构和生化数据，特别是小鼠 Rnr2 和酵母 Rnr2p?Rnr4p。目前还没有任何真核 Rnr1 结构的报道，我们使用 BioCARS 的 MAD 数据解析了酵母。 Rnr1 结构。在本提案中，我们将解决几个同源效应子-底物复合物结构，以解决如何通过在效应子位点结合的特定 dNTP 选择底物。酵母 RNR 组装涉及 Rnr2-Rnr4 与 Rnr1 的 C 端肽的关联。 Rnr4可以破坏这种组装，基于Rnr2肽设计一类新的抗癌和抗病毒抑制剂我们将解决。 Rnr1 与拟肽文库和抗癌药物（如氯法拉滨）复合的结构 在酵母中，RNR 活性由 ATP（上调因子）和 dATP（下调因子）进行变构控制。我们将解决 Rnr1 与 ATP 和 dATP 复合的结构，以解决这一问题。也被 Sml1 下调，Sml1 结合 Rnr1，抑制 RNR 活性并降低。我们将使用 MAD 和 Rnr1 与 Sml1 衍生肽和完整的 Sml1 复合来解析 Sml1 结构。我们构建了许多 Rnr1 突变体，以研究结构-功能关系。尽管人们普遍认为 Aβ 与阿尔茨海默病的发病机制有关，但我们将使用中和抗体 (Abs) 来获得其原子结构。单独的 Ab 可以与 AB 模型一起用于对接研究，或者更好的是，AB？Ab 复合物可以为设计抗阿尔茨海默病药物提供分子基础。我们已经以 apo 形式结晶了 AB 识别 Fab，并与截短的 A 复合。 ¿肽和其他几种 AB 识别抗体。