大肠杆菌DNA依赖的RNA聚合酶催化聚合反应的立体化学：用非对映异构ATP类似物进行的研究。
Stereochemistry of polymerization by DNA-dependent RNA-polymerase from Escherichia coli: an investigation with a diastereomeric ATP-analogue.
作者信息
Eckstein F, Armstrong V W, Sternbach H
出版信息
Proc Natl Acad Sci U S A. 1976 Sep;73(9):2987-90. doi: 10.1073/pnas.73.9.2987.
Abstract
The phosphodiester bond formation by DNA-dependent RNA-polymerase (RNA nucleotidyltransferase, nucleosidetriphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) can in principle result in retention, inversion, or racemization of configuration at the alpha-phosphorus of the nucleoside 5'-triphosphate being polymerized. As a first step in elucidating the stereochemistry of this reaction, one diastereomer (A) of adenosine 5'-O-(1-thiotriphosphate) (ATPalphaS) was polymerized with UTP in the presence of poly(dA-dT)-poly(dA-dT). The resulting polymer was enzymatically cleaved to uridine 2',3'-cyclic phosphorothioate which was determined to be the endo-isomer by comparison with an authentic sample. This shows that no reacemization had occurred and that isomer A of ATPalphaS gives a phosphorothioate diester bond with the R-configuration. Whether this represents inversion of retention of configuration awaits elucidation of the absolute configuration of isomer A for ATPalphaS.
摘要
依赖DNA的RNA聚合酶（RNA核苷酸转移酶，核苷三磷酸：RNA核苷酸转移酶，EC 2.7.7.6）形成磷酸二酯键原则上会导致正在聚合的核苷5'-三磷酸的α-磷处构型的保留、翻转或外消旋化。作为阐明该反应立体化学的第一步，在聚（dA-dT）-聚（dA-dT）存在下，将腺苷5'-O-（1-硫代三磷酸）（ATPαS）的一种非对映异构体（A）与UTP聚合。将所得聚合物酶切为尿苷2'，3'-环硫代磷酸酯，通过与真实样品比较确定其为内型异构体。这表明未发生外消旋化，并且ATPαS的异构体A形成具有R构型的硫代磷酸二酯键。这是代表构型翻转还是保留有待阐明ATPαS异构体A的绝对构型。
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