Studies on the nonmevalonate pathway to terpenes: The role of the GcpE (IspG) protein
Studies on the nonmevalonate pathway to terpenes: The role of the GcpE (IspG) protein
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DOI：
10.1073/pnas.201399298
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发表时间：
2001-12-18
影响因子：
11.1
通讯作者：
Rohdich, F
中科院分区：
文献类型：
--
作者：
Hecht, S;Eisenreich, W;Rohdich, F
为表达 xyIB 基因以及非甲羟戊酸途径基因而设计的重组大肠杆菌细胞提供有 C-13 标记的 1-脱氧-D-木酮糖。细胞提取物直接通过核磁共振波谱分析。在表达 xyIB、ispC、ispD、ispE 和 ispF 的细胞中检测到高水平的 C-13 标记的 2C-甲基-D-赤藓糖醇 2,4-环二磷酸。 gcpE 基因的额外表达提供了 1-羟基-2-甲基-2-(E)-丁烯基 4-二磷酸作为非甲羟戊酸途径的中间体。提出了涉及保守半胱氨酸残基的假设机制，用于在 GcpE 蛋白的催化下将 2C-甲基-D-赤藓糖醇 2,4-环二磷酸酶促转化为 1-羟基-2-甲基-2-(E)-丁烯基 4-二磷酸。
Recombinant Escherichia coli cells engineered for the expression of the xyIB gene in conjunction with genes of the nonmevalonate pathway were supplied with C-13-labeled 1-deoxy-D-xylulose. Cell extracts were analyzed directly by NMR spectroscopy. C-13-labeled 2C-methyl-D-erythritol 2,4-cyclodiphosphate was detected at high levels in cells expressing xyIB, ispC, ispD, ispE, and ispF. The additional expression of the gcpE gene afforded 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate as an intermediate of the nonmevalonate pathway. Hypothetical mechanisms involving conserved cysteine residues are proposed for the enzymatic conversion of 2C-methyl-D-erythritol 2,4-cyclodiphosphate into 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate catalyzed by the GcpE protein.