LIPIDOMIC ANALYSIS OF MYCOBACTERIUM TUBERCULOSIS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 结核分枝杆菌(Mtb)对主要药物产生抗药性，这加剧了结核病在全球的传播。迫切需要新的结核病疗法，但幸运的是，最近的基因组序列、遗传和蛋白质特征研究帮助确定了新的结核分枝杆菌药物靶点和战略干预的关键生化途径。在这方面，编码脂质代谢的基因是细菌基因组中非常重要的一部分，8%的基因组参与了这一活动。目前特别感兴趣的是结核分枝杆菌基因组中编码的多个细胞色素P450(P450)，其生物学作用尚不清楚。到目前为止，已经提出了细胞色素P125、细胞色素P142和细胞色素P51在甾醇代谢中的生理作用，以及细胞色素P132在脂肪酸代谢中的生理作用，但这些作用还没有确定。在这个项目中，我们打算通过对正常Mtb和单个P450酶被敲除的菌株的全局脂谱的比较分析来阐明Mtb P450酶的功能(S)，包括细胞色素P125、细胞色素P130和细胞色素P141。这种比较将使用傅里叶变换离子回旋共振质谱仪(FT-ICR MS)进行。这些P450的基因敲除菌株的可用性使得直接比较不同生长和标记条件下的脂肪体谱成为可能。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The global spread of tuberculosis (TB) has been aggravated by the development of strains of the causative bacterium Mycobacterium tuberculosis (Mtb) that are resistant to the leading drugs. New TB therapies are urgently needed, but fortunately recent genome sequence, genetic and protein characterization studies have helped identify novel Mtb drug targets and key biochemical pathways for strategic intervention. In this regard, genes that code for lipid metabolism are a very important part of the bacterial genome, and 8% of the genome is involved in this activity. Of particular interest in the present context are the multiple cytochromes P450 (P450) encoded in the Mtb genome, whose biological roles are not yet understood. To date, physiological roles have been proposed for CYP125 CYP142 and CYP51 in sterol metabolism and for CYP132 in fatty acid metabolism, but none of these roles has been established. In this project, we intend to elucidate the function(s) of Mtb P450 enzymes, including CYP125, CYP130 and CYP141, by comparative analyses of the global lipid profiles of normal Mtb and strains in which the individual P450 enzymes have been knocked out. This comparison will be carried out using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). The availability of knockout strains for these P450s makes possible a direct comparison of the lipidomic profiles under different growth and labeling conditions.