大肠杆菌组成型酶6-磷酸葡萄糖酸脱氢酶的体外合成
In vitro synthesis of a constitutive enzyme of Escherichia coli, 6-phosphogluconate dehydrogenase.
作者信息
Isturiz T, Wolf R E
出版信息
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4381-4. doi: 10.1073/pnas.72.11.4381.
Abstract
Enzymes of central intermediary metabolism are among the constitutive proteins of Escherichia coli. 6-Phosphogluconate dehydrogenase [6-phospho-D-gluconate: nicotinamide adenine dinucleotide phosphate 2-oxidoreductase (decarboxylating), EC 1.1.1.44-A1, an enzyme of the hexose monophosphate shunt, was synthesized in vitro in a coupled transcription-translation system directed by DNA from a specialized transducing phage carrying gnd, the structural gene. Enzyme synthesized in vitro was detected by radiochemical assay of its activity. Kinetic experiments using inhibitors of transcription and translation suggest that synthesis of the mRNA for the enzyme was initiated at the gnd promoter. Neither cyclic AMP nor cyclic GMP had any effect on the amount of enzyme synthesized in vitro.
摘要
中心中间代谢酶是大肠杆菌的组成蛋白之一。6-磷酸葡萄糖酸脱氢酶[6-磷酸-D-葡萄糖酸：烟酰胺腺嘌呤二核苷酸磷酸2-氧化还原酶（脱羧），EC 1.1.1.44-A1，一种磷酸己糖支路的酶]，是在一个由携带gnd（结构基因）的特异性转导噬菌体的DNA指导的体外偶联转录-翻译系统中合成的。通过对其活性的放射化学测定来检测体外合成的酶。使用转录和翻译抑制剂的动力学实验表明，该酶的mRNA合成起始于gnd启动子。环磷酸腺苷和环磷酸鸟苷对体外合成的酶量均无任何影响。
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