Doublié S, Tabor S, Long A M, Richardson C C, Ellenberger T
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Nature. 1998 Jan 15;391(6664):251-8. doi: 10.1038/34593.
DNA polymerases change their specificity for nucleotide substrates with each catalytic cycle, while achieving error frequencies in the range of 10(-5) to 10(-6). Here we present a 2.2 A crystal structure of the replicative DNA polymerase from bacteriophage T7 complexed with a primer-template and a nucleoside triphosphate in the polymerase active site. The structure illustrates how nucleotides are selected in a template-directed manner, and provides a structural basis for a metal-assisted mechanism of phosphoryl transfer by a large group of related polymerases.
DNA聚合酶在每个催化循环中都会改变其对核苷酸底物的特异性,同时实现10^(-5)至10^(-6)范围内的错误频率。在此,我们展示了噬菌体T7复制性DNA聚合酶与引物模板以及位于聚合酶活性位点的三磷酸核苷复合的2.2埃晶体结构。该结构阐明了核苷酸如何以模板导向的方式被选择,并为一大类相关聚合酶的磷酸转移金属辅助机制提供了结构基础。