大肠杆菌DNA聚合酶I大片段（克列诺片段）空转反应的机制。
Mechanism of the idling-turnover reaction of the large (Klenow) fragment of Escherichia coli DNA polymerase I.
作者信息
Mizrahi V, Benkovic P A, Benkovic S J
出版信息
Proc Natl Acad Sci U S A. 1986 Jan;83(2):231-5. doi: 10.1073/pnas.83.2.231.
Abstract
The mechanism of the idling-turnover reaction catalyzed by the large (Klenow) fragment of Escherichia coli DNA polymerase I has been investigated. The reaction cycle involved is one of excision/incorporation, in which the 3' deoxynucleotide residue of the primer DNA strand is partitioned into its 5'-mono- and 5'-triphosphate derivatives, respectively. Mechanistic studies suggest the 5'-monophosphate product is formed in the first step by simple 3'----5' exonucleolytic cleavage. Rapid polymerization follows with the concomitant release of inorganic pyrophosphate. In the second step, the 5'-triphosphate product is generated by a pyrophosphorolysis reaction, which, despite the low concentration of pyrophosphate that has accumulated, occurs at a rate that is comparable with that of the parallel 3'----5' hydrolysis reaction.
摘要
对大肠杆菌DNA聚合酶I大片段（克列诺片段）催化的空转反应机制进行了研究。所涉及的反应循环是切除/掺入循环之一，其中引物DNA链的3'脱氧核苷酸残基分别被分割成其5'-单磷酸和5'-三磷酸衍生物。机理研究表明，5'-单磷酸产物在第一步通过简单的3'→5'核酸外切酶切割形成。随后快速聚合，同时释放无机焦磷酸。在第二步中，5'-三磷酸产物通过焦磷酸解反应生成，尽管积累的焦磷酸浓度很低，但该反应的发生速率与平行的3'→5'水解反应相当。
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