Assays and Screens

该项目的目标是开发分析方法并确定将用作研究工具的化合物 结核分枝杆菌(Mtb)的研究，以及作为治疗结核病(TB)的潜在药物。它是 预计这一方法将向管道提供化验和化学投入，用于开发 用于治疗结核病感染和结核分枝杆菌研究的新的领导分子。通过以下途径鉴定的化合物 本项目的化验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。 该项目将重点放在基本Mtb膜蛋白的初始目标清单上，选择用于其 功能重要性，为本计划项目开发。具体目标是：(1)发展生物化学 检测Mtb膜蛋白并高通量筛选小分子文库，(2) 利用胶束中的Mtb蛋白进行小分子文库的核磁共振筛选，以及(3)开发和 进行双层和双分子中Mtb蛋白小分子文库的核磁共振筛选。 将开发生化分析和基于核磁共振的筛选方法。生化化验将会 促进进一步的功能鉴定，并能够筛选干扰 活动。核磁共振将被用来筛选甚至在蛋白质之前就与结核分枝杆菌蛋白质结合的小分子。 函数有完整的注释。通过这种方法识别弱粘结剂对两个方面都非常有用 项目1和项目3中的功能和结构确定过程，并有可能识别药物- 比如用于治疗开发的先导化合物。对于球状蛋白质的核磁共振筛选是非常成熟的 水溶液，但尚未被应用于胶束中的膜蛋白或 脂质双层，并开发在脂质环境中膜蛋白的核磁共振筛选是一个主要目标 这个项目。

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.