M. tuberculosis Cell Wall Biogenesis; New Drugs; TB-HIV

描述（由申请人提供）： 自艾滋病开始流行以来，与艾滋病毒有关的结核病的发病率在全世界一直在增加，预计今后还会进一步增加，特别是在发展中国家。艾滋病毒感染的加速和放大影响导致结核分枝杆菌多药耐药菌株引起的疾病发病率增加。因此，开发抗结核病的新药对于控制这两种感染都很重要。由于分枝杆菌细胞壁在分枝杆菌表面形成几乎不可渗透的保护性屏障，因此分枝杆菌细胞壁是合理设计抗结核药物的有吸引力的靶点。目前用于治疗TB的一些最有效的药物影响其主链的组分-分枝杆菌氨基半乳聚糖-肽聚糖（mAGP）复合物。我们的长期目标是确定参与mAGP组装的过程和酶。最近在M.因此，这项拨款建议的具体目的是：1。通过克隆、过表达和后续生化鉴定，鉴定AG生物合成簇中参与分枝杆菌半乳聚糖生物合成的基因，并确定其生物学功能。2.通过突变体/条件突变体的制备和表型表征，确定AG生物合成簇内推定ABC转运蛋白的功能。该方法将有助于确定微生物生物化学中更复杂的途径之一，并揭示可用于药物开发的反应。该研究将主要在斯洛伐克布拉迪斯拉发的夸美纽斯大学自然科学学院与Katarina Mikusova合作进行，作为NIH资助A1-18357的延伸。

DESCRIPTION (provided by applicant): The incidence of HIV-associated tuberculosis has been increasing worldwide since the beginning of the AIDS epidemic, and is expected to rise even further in the future, especially in developing countries. The accelerating and amplifying influence of HIV infection is contributing to the increasing incidence of disease caused by multidrug-resistant strains of Mycobacterium tuberculosis. Development of new drugs against tuberculosis is thus important for control of both of the infections. Mycobacterial cell wall is an attractive target for rational drug design against tuberculosis, due to the fact that it forms a protective, almost impermeable barrier, on the surface of mycobacteria. Some of the most effective drugs currently used for the treatment of TB affect components of its backbone - mycolylarabinogalactan-peptidoglycan (mAGP) complex. Our long-term goal is to identify processes and enzymes involved in the mAGP assembly. Possible AG biosynthetic gene cluster has been recently identified in the genome of M. tuberculosis and thus the specific aims of this grant proposal are: 1. Identify the genes involved in mycobacterial galactan biosynthesis within AG biosynthetic cluster and determine their biological functions via cloning, overexpression and subsequent biochemical characterization. 2. Establish the function of the putative ABC transporter within the AG biosynthetic cluster by way of preparation and phenotypic characterization of the mutants/conditional mutants. The approach will help define one of the more complex pathways in microbial biochemistry and reveal reactions that should be exploitable for drug development. The research will be primarily carried out at Comenius University, Faculty of Natural Sciences in Bratislava, Slovakia in collaboration with Katarina Mikusova, as an extension of the NIH grant A1-18357.