Core C: Enzymology Core

核心 C：酶学核心

• 批准号: 10513682
• 负责人: CRAIG E. CAMERON
• 金额: $ 746.26万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-16 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10513682
• 关键词: Alberta province; Alphavirus; Bacteria; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biophysics; COVID-19 assay; Cells; Collaborations; Communities; Complex; Coronavirus; Cryoelectron Microscopy; Development; Distant; Ebola virus; Enzymatic Biochemistry; Enzyme Inhibition; Enzyme Kinetics; Enzymes; Exhibits; Filovirus; Flavivirus; Generations; Goals; HIV-1; Insecta; International; Laboratories; Lead; Mammalian Cell; Measurement; Miniaturization; Monitor; Multienzyme Complexes; Multiprotein Complexes; Mutation; Nucleotides; Polymerase; Process; Protein Family; RNA Helicase; RNA-Directed DNA Polymerase; Records; Research Project Grants; Resistance; Resistance development; Reverse Transcriptase Inhibitors; SARS coronavirus; Structure; System; Technology; Testing; Translations; Universities; Validation; Viral; Virus; X-Ray Crystallography; antiviral drug development; biophysical techniques; density; drug development; drug discovery; high throughput screening; inhibitor; insight; interest; lead optimization; member; programs; protein expression; protein purification; protocol development; screening; single molecule; structural biology; three dimensional structure; tool; virology

ABSTRACT The overarching goal of Core C is to support READDI-AC discovery efforts in collaboration with Discovery Core B and MedChem Core D by validation, characterization, and optimization of compounds in enzyme assays and structural studies, and to provide the Research Projects with detailed mechanisms of action and mechanistic insights to mitigate potential problems with resistance conferring mutations. Enzymology Core C includes seven internationally recognized laboratories that cover complementary expertise at the interfaces of virology, biochemistry, biophysics and structural biology. Three specific aims that describe the central activities of Enzymology Core C: Aim 1: Develop assays to support hit discovery efforts by Discovery Core B. Key activities include the expression and purification of enzymes that the Research Projects and Discovery Core B have selected as targets. Enzymology Core C will support the development of protocols for the expression and purification of new enzyme targets and their corresponding assays to monitor activity and inhibition. The translation of enzyme assays into high density formats to facilitate high throughput screening by Discovery Core B is a second responsibility. Aim 2. Structural Biology to guide hit and lead optimization. Enzymology Core C will focus on solving 3D structures of polymerases and RNA helicases that are currently not available to the scientific community. Priority will also be given to solving the structures of flavivirus and alphavirus polymerases in complex with the nucleotide and non-nucleotide analog inhibitors to provide detailed information on the inhibitor binding sites. Aim 3. Support the Projects in detailed mechanism of action studies for lead compounds. Functional assays will be employed to assess the inhibitory activity against the biologically relevant enzyme target, such as measurements of the half maximal inhibitory concentration to determine compound potency. Promising hits and leads will also be tested against homologous enzymes within the protein family as well as distant enzymes to assess breadth and the potential for off-target activity. Enzymology Core C will provide an array of state-of-the-art biochemical/biophysical approaches to provide MedChem Core D and the Projects with information on the mechanisms associated with inhibition and resistance.

摘要 Core C的总体目标是与Discovery Core合作支持Readdi-AC发现工作 B和MedChem Core D通过酶分析中化合物的验证、表征和优化以及 结构研究，并为研究项目提供详细的作用机制和机制 对缓解导致耐药性突变的潜在问题的见解。 酶学核心C包括七个国际公认的实验室，涵盖互补的专业知识 在病毒学、生物化学、生物物理学和结构生物学的交界处。描述以下三个具体目标 酶学核心C的中心活动： 目标1：开发测试以支持Discovery Core B的热门发现工作。 主要活动包括研究项目和发现的酶的表达和纯化 核心B已被选为目标。酶学核心C将支持开发用于 新的酶靶标的表达和纯化及其活性监测方法 抑制力。将酶分析转换为高密度格式，以通过以下方式促进高通量筛选 发现核心B是第二项责任。 目的2.用结构生物学来指导命中和引导优化。 酶学核心C将专注于解决聚合酶和RNA解旋酶的3D结构，这些结构目前 科学界无法获得。还将优先解决黄病毒的结构和 在甲型病毒聚合酶与核苷酸和非核苷酸类似物的复合体中提供了详细的 关于抑制剂结合位点的信息。 目的3.支持先导化合物的详细作用机理研究项目。 功能分析将被用来评估对生物相关酶的抑制活性。 靶标，如测量半数最大抑菌浓度，以确定复合效力。 有希望的命中和线索也将与蛋白质家族中的同源酶以及 远距离酶，以评估广度和潜在的偏离目标的活动。酶学核心C将提供一种 一系列最先进的生化/生物物理方法，为MedChem Core D和项目提供 关于与抑制和抗性相关的机制的信息。