Eoh Hyungjin, Narayanasamy Prabagaran, Brown Amanda C, Parish Tanya, Brennan Patrick J, Crick Dean C
Mycobacteria Research Laboratories, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523, USA.
Chem Biol. 2009 Dec 24;16(12):1230-9. doi: 10.1016/j.chembiol.2009.10.014.
Many bacterial pathogens utilize the 2-C-methyl-D-erythritol 4-phosphate pathway for biosynthesizing isoprenoid precursors, a pathway that is vital for bacterial survival and absent from human cells, providing a potential source of drug targets. However, the characterization of 4-diphosphocytidyl-2-C-methyl-D-erythritol (CDP-ME) kinase (IspE) has been hindered due to a lack of enantiopure CDP-ME and difficulty in obtaining pure IspE. Here, enantiopure CDP-ME was chemically synthesized and recombinant IspE from bacterial pathogens were purified and characterized. Although gene disruption was not possible in Mycobacterium tuberculosis, IspE is essential in Mycobacterium smegmatis. The biochemical and kinetic characteristics of IspE provide the basis for development of a high throughput screen and structural characterization.
许多细菌病原体利用2-C-甲基-D-赤藓糖醇4-磷酸途径来生物合成类异戊二烯前体，该途径对细菌生存至关重要且在人类细胞中不存在，是潜在的药物靶点来源。然而，由于缺乏对映体纯的4-二磷酸胞苷-2-C-甲基-D-赤藓糖醇（CDP-ME）以及难以获得纯的IspE，4-二磷酸胞苷-2-C-甲基-D-赤藓糖醇激酶（IspE）的特性研究受到了阻碍。在此，通过化学合成获得了对映体纯的CDP-ME，并对来自细菌病原体的重组IspE进行了纯化和特性分析。虽然在结核分枝杆菌中无法进行基因破坏，但IspE在耻垢分枝杆菌中是必需的。IspE的生化和动力学特性为高通量筛选和结构表征的发展提供了基础。
