LIPIDOMIC ANALYSIS OF MYCOBACTERIUM TUBERCULOSIS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 结核病（TB）的全球传播由于对主要药物具有耐药性的致病细菌结核分枝杆菌（Mtb）菌株的发展而加剧。迫切需要新的结核病疗法，但幸运的是，最近的基因组序列，遗传和蛋白质表征研究有助于确定新的结核病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。在本上下文中特别感兴趣的是Mtb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。迄今为止，已经提出了CYP 125、CYP 142和CYP 51在固醇代谢中的生理作用，以及CYP 132在脂肪酸代谢中的生理作用，但这些作用都尚未确定。在这个项目中，我们打算阐明结核分枝杆菌P450酶，包括CYP 125，CYP 130和CYP 141的功能，通过比较分析正常结核分枝杆菌和菌株的整体脂质谱，其中个别P450酶已被敲除。将使用傅里叶变换离子回旋共振质谱法（FT-ICR MS）进行比较。这些P450的敲除菌株的可用性使得在不同生长和标记条件下直接比较脂质组学特征成为可能。这项工作得到了NIH资助AI 074824的支持。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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. This work is supported by NIH grant AI074824.