Crystallization of the Galactosyltransferase from Mtb

Mtb 半乳糖基转移酶的结晶

• 批准号: 6571603
• 负责人: Robert C Reynolds
• 金额: $ 21.32万
• 依托单位: SOUTHERN RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-15 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6571603
• 关键词: Escherichia coli; Mycobacterium tuberculosis; active sites; antitubercular agents; carbohydrate metabolism; crystallization; drug design /synthesis /production; galactans; galactosyltransferases; gene expression; monosaccharides; multidrug resistance; protein folding; protein purification; protein structure; recombinant proteins

DESCRIPTION (provided by applicant): This application is in response to PAS-02-031 for a supplement to our current NIAID-funded program AI45317 entitled Glycosyltransferases as Drug Targets in Mycobacteria. This supplemental program will involve the expression, purification, and crystallization of the galactosyltransferase (Rv3808c) from Mycobacterium tuberculosis. The galactosyltransferase adds galactofuranose units to the growing galactan, a polysaccharide that anchors the mycolylarabinan superstructure and is critical to mycobacterial cell wall integrity in the harsh environment of the human macrophage. This effort is responsive to the announcement in that it supplements our current program by applying modern structural biology techniques to obtain a structure of a protein targeted for drug development under our current grant. This protein was not available at the time of our application, and the information provided through the application of the requested supplemental funds will give us a specific picture of the protein active site allowing more rapid development of drugs to target this critical mycobacterial enzyme. As well, this grant is responsive to the PA in that it addresses a new drug target in tuberculosis, and may have implications in the treatment of multi-drug resistant tuberculosis (MDR-TB), a critical health problem worldwide. Tuberculosis and its drug resistant forms are the focus of two active NIAID Program Announcements (PA-99-124 & PA-01-113) and is considered a potential biological weapon by the Department of Defense. This application involves modern structural biology that may not be considered as innovative as recent techniques in bioinformatics, genomics, and microarray technology. This effort, however, is innovative in the choice of target and the impact that structural biology can have on our funded program.

描述（由申请人提供）：此申请是针对PAS-02-031的响应，以补充我们当前的NIAID资助计划AI45317标题为糖基转移酶是分枝杆菌中的药物靶标。该补充程序将涉及结核分枝杆菌的半乳糖基转移酶（RV3808C）的表达，纯化和结晶。半乳糖基转移酶为生长的甲乳转糖糖添加了半乳核素单元，这是一种锚定霉菌甲拉比诺式上层结构的多糖，对人类巨噬细胞的恶劣环境中的分枝杆菌细胞壁完整性至关重要。这项努力对公告的反应迅速，因为它通过应用现代结构生物学技术来补充我们当前的计划，以获得针对药物开发的蛋白质结构，该结构是我们当前赠款的。该蛋白在我们应用时无法获得，并且通过应用所需的补充资金提供的信息将为我们提供蛋白质活性位点的特定图片，从而使药物的开发更快地针对这种关键的分枝杆菌酶。同样，该赠款对PA的响应敏感，因为它解决了结核病的新药物靶标，并且可能对全世界关键健康问题的多药耐药性结核病（MDR-TB）有影响。结核病及其耐药性形式是两个活跃的NIAID计划公告（PA-99-124＆PA-01-113）的重点，并被国防部视为潜在的生物武器。该应用涉及现代结构生物学，这些生物学可能不像生物信息学，基因组学和微阵列技术中的最新技术一样具有创新性。但是，这项努力在选择目标以及结构生物学对我们资助的计划的影响方面具有创新性。