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: 7956842
• 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/7956842
• 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.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 核糖核苷酸还原（RNR）通过将NDPS还原为DNDP来催化从头DNA合成的速率限制步骤。尽管存在许多关于真核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，但到目前为止，尚无任何真核RNR1的结构。使用来自biocar的疯狂数据，我们解决了酵母RNR1结构。在此提案中，我们将解决几个同源效应子基底复合物结构，以解决效应点上特定的DNTP结合选择底物的方式。酵母RNR组件涉及RNR2-RNR4与RNR1的关联。 RNR2和RNR4的C末端肽会破坏该组件，并且基于RNR2 Pepperides设计了新的抗癌和抗病毒抑制剂。我们将求解与肽拟合文库和抗癌剂（如克叶曲滨）复合的RNR1结构。在酵母中，RNR活动由ATP（上调）和DatP（下调器）通过变构控制。我们将解决与ATP和DATP复合的RNR1的结构来解决此问题。酵母RNR也被SML1下调，该SML1结合RNR1。 SML1的C末端以降低的效力抑制RNR活性。我们将使用与SML1衍生的肽和完整SML1复合的MAD和RNR1解决SML1结构。我们已经构建了RNR1的许多突变体研究结构 - 功能关系。有几个以天然形式和效应 - 底物复合物的复杂形式结晶。尽管接受Abeta参与了阿尔茨海默氏症的发病机理，但其原子结构尚无共识。我们将使用中和抗体（ABS）获得此功能。仅ABS的结构可以与AB模型一起用于对接研究，或者更好的是，AB？AB复合物可以为针对阿尔茨海默氏病设计药物提供分子基础。我们已经以Apo形式结晶了AB识别的Fab，并将其复合到截短的A肽和其他几种AB识别的ABS中。