Crystallization of the Galactosyltransferase from Mtb

描述(申请人提供)：本申请是为了响应PAS-02-031对我们目前由NIAID资助的名为糖基转移酶作为分枝杆菌药物靶标的项目AI45317的补充。这个补充计划将涉及结核分枝杆菌半乳糖基转移酶(Rv3808c)的表达、纯化和结晶。半乳糖基转移酶将呋喃半乳糖单位添加到正在生长的半乳聚糖中，半乳聚糖是一种锚定分枝杆菌超微结构的多糖，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。这一努力是对这一宣布的回应，因为它通过应用现代结构生物学技术来获得我们目前拨款下用于药物开发的蛋白质的结构，从而补充了我们目前的计划。在我们申请时，这种蛋白质还无法获得，通过申请补充资金提供的信息将使我们对蛋白质活性部位有一个具体的了解，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的响应，因为它针对的是结核病的一个新的药物靶标，并可能对耐多药结核病(MDR-TB)的治疗产生影响，MDR-TB是世界范围内的一个关键健康问题。结核病及其耐药形式是NIAID两个现行计划公告(PA-99-124和PA-01-113)的重点，并被国防部视为潜在的生物武器。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中的最新技术那样具有创新性。然而，这一努力在目标的选择和结构生物学可能对我们资助的项目产生的影响方面是创新的。

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.