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: 7601582
• 负责人: Chris G Dealwis
• 金额: $ 2.75万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601582
• 关键词: Address; Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Antineoplastic Agents; Antiviral Agents; Ants; Binding; Biochemical; C-terminal; Catalytic Domain; Class; Clofarabine; Complex; Computer Retrieval of Information on Scientific Projects Database; Consensus; DNA biosynthesis; DNA chemical synthesis; Data; Docking; Drug Design; Funding; Grant; Institution; Libraries; Modeling; Molecular; Mus; Pathogenesis; Peptides; Rate; 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

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.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 核糖核苷酸还原酶(RNR)通过将NDPs还原为dNDPs来催化从头合成DNA的限速步骤。虽然有许多关于真核细胞Rnr1非催化亚基的结构和生化数据，特别是小鼠Rnr2和酵母Rnr2p和Rnr4p，但到目前为止还没有关于真核细胞Rnr1结构的报道。利用BioCARS的MAD数据，我们解析了酵母Rnr1的结构。在这个提案中，我们将解决几个同源的效应器-底物复杂结构，以解决如何通过特定的dNTPs结合在效应器位置来选择底物。酵母RNR组装涉及Rnr2-Rnr4与Rnr1的结合。Rnr2和Rnr4的C端肽可以破坏这种组装，在Rnr2多肽的基础上设计了一类新的抗癌和抗病毒抑制剂。我们将解决Rnr1与模拟多肽文库和抗癌药物如氯法拉滨的复合结构。在酵母中，RNR活性受ATP(上调因子)和dATP(下调调节因子)的变构控制。我们将解决Rnr1与ATP和dATP的复合结构来解决这个问题。酵母RNR也被Sml1下调，Sml1与Rnr1结合。Sml1的C末端抑制RNR活性，但效力降低。我们将使用MAD和Rnr1与Sml1衍生的多肽和完整的Sml1复合来解决Sml1的结构。我们已经构建了大量的Rnr1突变体来研究结构与功能的关系。有几个已经结晶为天然形式和与效应器-底物络合物的络合物。虽然公认阿贝塔与阿尔茨海默病的发病机制有关，但对于它的原子结构还没有达成共识。我们将使用中和抗体(Abs)来获得这一点。这些抗体的结构可以单独用于AB模型的对接研究，或者更好的是，AB？抗体复合体可以为设计抗阿尔茨海默病药物提供分子基础？S。我们以载脂蛋白的形式结晶了一个识别AB的Fab，并将其络合成一个截短的A？肽，以及其他几个识别AB的抗体。