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.

抽象的 核心C的总体目标是支持与Discovery Core合作的ReadDi-AC发现工作 B和Medchem核心D通过酶测定中化合物的验证，表征和优化和优化 结构研究，并为研究项目提供详细的作用机制和机械机制 洞察力减轻抗性赋予突变的潜在问题。 酶学核心C包括七个涵盖互补专业知识的国际认可的实验室 在病毒学，生物化学，生物物理学和结构生物学的界面上。描述的三个特定目标 酶学核心C的中心活动C： 目的1：开发测定法，以支持Discovery Core B的HIT发现工作。 关键活动包括研究项目和发现的酶的表达和纯化 核心B已选择为目标。酶学核心C将支持制定协议 新酶靶标及其相应测定的表达和纯化，以监测活动和 抑制。将酶测定转换为高密度格式，以促进高吞吐量筛选 发现核心B是第二个责任。 目标2。指导命中和铅优化的结构生物学。 酶学核心C将重点求解目前是聚合酶和RNA解旋酶的3D结构 科学界无法使用。还将优先考虑解决黄体病毒的结构和 与核苷酸和非核苷酸类似物抑制剂复合物中的α病毒聚合酶可提供详细的 有关抑制剂结合位点的信息。 目标3。支持铅化合物的详细行动研究机理。 功能测定将用于评估针对生物学相关酶的抑制活性 靶标，例如测量半最大抑制浓度以确定复合效力。 有希望的命中和铅也将针对蛋白质家族中的同源酶进行测试 遥远的酶评估广度和脱靶活动的潜力。酶学核心C将提供 一系列最先进的生化/生物物理方法，以提供Medchem Core D和项目 有关与抑制和抗性相关的机制的信息。