Shan Shan, Chen Xuehui
Structural Biology Laboratory, Tsinghua University, Beijing 100084, People's Republic of China.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Jul 1;67(Pt 7):821-3. doi: 10.1107/S1744309111019567. Epub 2011 Jun 30.
The 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase (IspE) from Mycobacterium tuberculosis, an enzyme from the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, is crucial and essential for the survival of this pathogenic bacterium. IspE catalyzes the conversion of 4-diphosphocytidyl-2-C-methyl-D-erythritol (CDP-ME) to 4-diphosphocytidyl-2-C-methyl-D-erythritol 2-phosphate (CDP-ME2P) in an ATP-dependent manner. Solving the crystal structure of M. tuberculosis IspE will shed light on its structural details and mechanism of action and may provide the basis for the future design of drugs for the treatment of multidrug-resistant and extremely drug-resistant M. tuberculosis strains. Recombinant M. tuberculosis IspE was crystallized at 291 K using NaCl or Li2SO4 as a precipitant. A 2.1 Å resolution native data set was collected from a single flash-cooled crystal (100 K) belonging to space group P2(1)2(1)2(1), with unit-cell parameters a=52.5, b=72.3, c=107.3 Å. One molecule was assumed per asymmetric unit, which gives a Matthews coefficient of 3.4 Å3 Da(-1) with 63% solvent content.
结核分枝杆菌的4-二磷酸胞苷-2-C-甲基-D-赤藓醇激酶（IspE）是2-C-甲基-D-赤藓醇4-磷酸（MEP）途径中的一种酶，对这种致病细菌的存活至关重要且必不可少。IspE以ATP依赖的方式催化4-二磷酸胞苷-2-C-甲基-D-赤藓醇（CDP-ME）转化为4-二磷酸胞苷-2-C-甲基-D-赤藓醇2-磷酸（CDP-ME2P）。解析结核分枝杆菌IspE的晶体结构将揭示其结构细节和作用机制，并可能为未来设计治疗耐多药和广泛耐药结核分枝杆菌菌株的药物提供依据。使用NaCl或Li2SO4作为沉淀剂，在291 K下使重组结核分枝杆菌IspE结晶。从属于空间群P2(1)2(1)2(1)、晶胞参数a=52.5、b=72.3、c=107.3 Å的单个快速冷却晶体（100 K）收集到分辨率为2.1 Å的天然数据集。每个不对称单元假定有一个分子，这给出了马修斯系数为3.4 Å3 Da(-1)，溶剂含量为63%。
