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）：本申请是对PAS-02-031的回应，作为我们目前niaid资助的项目AI45317的补充，题为“分枝杆菌中糖基转移酶作为药物靶点”。本补充项目将涉及结核分枝杆菌半乳糖转移酶（Rv3808c）的表达、纯化和结晶。半乳糖转移酶将半乳糖呋喃糖单位添加到生长的半乳聚糖中，半乳聚糖是一种锚定菌胞糖聚糖上层结构的多糖，在人类巨噬细胞的恶劣环境中对分枝杆菌细胞壁的完整性至关重要。这项工作是对公告的回应，因为它通过应用现代结构生物学技术来获得我们当前资助下药物开发目标蛋白质的结构，从而补充了我们当前的项目。在我们申请时，这种蛋白质是不可用的，通过申请所要求的补充资金提供的信息将为我们提供蛋白质活性位点的具体图片，从而更快地开发针对这种关键分枝杆菌酶的药物。此外，这笔赠款是对PA的回应，因为它针对结核病的一个新的药物靶点，并可能对治疗耐多药结核病（MDR-TB）产生影响，这是一个全球性的严重卫生问题。结核病及其耐药形式是两个NIAID项目公告（PA-99-124和PA-01-113）的重点，被国防部认为是一种潜在的生物武器。这一应用涉及现代结构生物学，可能不像生物信息学、基因组学和微阵列技术那样具有创新性。然而，这项工作在目标的选择和结构生物学对我们资助的项目的影响方面是创新的。