The purification, crystallization and structural elucidation of dTDP-D-glucose 4,6-dehydratase (RmIB), the second enzyme of the dTDP-L-rhamnose synthesis pathway from Salmonella enterica serovar Typhimurium
The purification, crystallization and structural elucidation of dTDP-D-glucose 4,6-dehydratase (RmIB), the second enzyme of the dTDP-L-rhamnose synthesis pathway from Salmonella enterica serovar Typhimurium
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DOI：
10.1107/s0907444999016200
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发表时间：
2000-02-01
影响因子：
2.2
通讯作者：
Naismith, JH
中科院分区：
文献类型：
--
作者：
Allard, STM;Giraud, MF;Naismith, JH
dTDP-D-葡萄糖4，6-脱氢酶（RmlB）是参与dTDP-L-鼠李糖途径的四种酶中的第二种，并催化dTDP-D-葡萄糖脱水为dTDP-4-酮-6-脱氧-D-葡萄糖。该途径的最终产物dTDP-L-鼠李糖是L-鼠李糖的前体，L-鼠李糖是许多病原菌细胞壁的关键成分。来自沙门氏菌鼠伤寒血清型的RmlB已被过表达和纯化，并且酶的晶体已使用以硫酸锂作为沉淀剂的坐滴蒸气扩散技术生长。在一个空间群为P2（1）的单晶Rm 1 B上获得了分辨率为2.8埃的衍射数据，晶胞参数a = 111.85，B = 87.77，c = 145.66埃，β = 131.53度。不对称单元含有四种单体，呈两种RmlB二聚体形式，溶剂含量为62%。已经获得了分子置换解决方案，目前正在完善模型。
dTDP-D-glucose 4,6-dehydratase (RmlB) is the second of four enzymes involved in the dTDP-L-rhamnose pathway and catalyzes the dehydration of dTDP-D-glucose to dTDP-4-keto-6-deoxy-D-glucose. The ultimate product of the pathway, dTDP-L-rhamnose, is the precursor of L-rhamnose, which is a key component of the cell wall of many pathogenic bacteria. RmlB from Salmonella enterica serovar Typhimurium has been overexpressed and purified, and crystals of the enzyme have been grown using the sitting-drop vapour-diffusion technique with lithium sulfate as precipitant. Diffraction data have been obtained to a resolution of 2.8 Angstrom on a single frozen RmlB crystal which belongs to space group P2(1), with unit-cell parameters a = 111.85, b = 87.77, c = 145.66 Angstrom, beta = 131.53 degrees. The asymmetric unit contains four monomers in the form of two RmlB dimers with a solvent content of 62%. A molecular-replacement solution has been obtained and the model is currently being refined.