Assays and Screens

测定和筛选

• 批准号: 8380951
• 负责人: Francesca M Marassi
• 金额: $ 38.94万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8380951
• 关键词: Address; Algorithms; Antitubercular Agents; Automation; Binding; Biochemical; Biological; Biological Assay; Biological Process; Biology; California; Chemicals; Development; Drug Delivery Systems; Engineering; Environment; Enzyme-Linked Immunosorbent Assay; Fluorescence; Goals; Image; Institutes; Label; Laboratories; Lead; Ligands; Lipid Bilayers; Lipids; Medical Research; Membrane; Membrane Proteins; Methods; Micelles; Mycobacterium tuberculosis; Organism; Pharmaceutical Preparations; Pharmacologic Substance; Preparation; Principal Investigator; Process; Production; Proteins; Protocols documentation; Research; Sampling; Screening procedure; Solutions; Structure; Technology; Transmembrane Transport; Transport Process; Tuberculosis; Universities; Validation; aqueous; assay development; base; drug development; experience; globular protein; high throughput screening; insight; instrumentation; luminescence; magnetic field; mycobacterial; programs; protein function; small molecule; small molecule libraries; solid state nuclear magnetic resonance; therapeutic development; tool; tuberculosis drugs

The goal of this Project is to develop assays and identify compounds that will be useful as research tools for studies of Mycobacterium tuberculosis (Mtb), and as potential drugs against tuberculosis (TB). It is anticipated that this approach will provide assay and chemical inputs into the pipeline for the development of new lead molecules for the treatment of TB infection, and for Mtb research. Compounds identified through the assay development and screening efforts of this Project, will be useful as research tools, providing important insights about the biological functions and structures of targeted Mtb membrane proteins, and will may be useful for therapeutic developments. This Project will focus on an Initial Target List of essential Mtb membrane proteins, selected for their functional importance, developed for this Program Project. The specific aims are (1) to develop biochemical assays for Mtb membrane proteins and perform high throughput screening of small molecule libraries, (2) to perform NMR screening of small molecule libraries with the Mtb proteins in micelles, and (3) to develop and perform NMR screening of small molecule libraries for the Mtb proteins in bilayers and bicelles. Biochemical assays and NMR-based methods for screening will be developed. The biochemical assays will facilitate further functional characterization, and enable the screening of compounds that interfere with activity. NMR will be used to screen for small molecules that bind to Mtb proteins even before a protein function is fully annotated. The identification of weak binders by this method is very useful for both the function and structure determination processes in Projects 1 and 3, and has the potential of identifying drug- like lead compounds for therapeutic development. NMR screening is well developed for globular proteins in aqueous solutions, but it has not been applied to membrane proteins in micelles or membrane proteins in lipid bilayers, and developing NMR screening for membrane proteins in lipid environments is a major goal of this Project.

该项目的目的是开发测定法和确定将作为研究工具有用的化合物 结核分枝杆菌（MTB）的研究以及针对结核病（TB）的潜在药物。这是 预计这种方法将为开发的管道提供测定和化学输入 用于治疗结核病感染和MTB研究的新铅分子。通过确定的化合物 该项目的测定开发和筛选工作将作为研究工具有用，提供 关于靶向MTB膜蛋白的生物学功能和结构的重要见解，并将 可能对治疗发展有用。 该项目将重点放在必需的MTB膜蛋白的初始目标列表上 功能重要性，为该计划项目开发。具体目的是（1）开发生化 MTB膜蛋白的测定法，对小分子文库进行高吞吐量筛选，（2）至 用胶束中的MTB蛋白对小分子库进行NMR筛选，（3）进行和（3） 对双层和双层中的MTB蛋白进行小分子库进行NMR筛选。 将开发生化化学测定和基于NMR的筛查方法。生化测定将 促进进一步的功能表征，并能够筛选干扰的化合物 活动。 NMR将用于筛选与MTB蛋白结合的小分子，甚至在蛋白质之前 功能已完全注释。通过这种方法识别弱粘合剂对这两个方法都非常有用 项目1和3中的功能和结构确定过程，并具有识别药物的潜力 就像用于治疗开发的铅化合物一样。 NMR筛选针对球状蛋白的发展良好 水溶液，但尚未应用于胶束或膜蛋白中的膜蛋白 脂质双层和在脂质环境中开发膜蛋白的NMR筛选是一个主要目标 这个项目。