Structures of KlenTaq DNA Polymerase Caught While Incorporating C5-Modified Pyrimidine and C7-Modified 7-Deazapurine Nucleoside Triphosphates

被引：69

作者：

Bergen, Konrad

Steck, Anna-Lena

Struett, Stefan

Baccaro, Anna

Welte, Wolfram

Diederichs, Kay

Marx, Andreas ^{[1
]}

机构：

[1] Univ Konstanz, Konstanz Res Sch Chem Biol, Dept Chem, D-78457 Constance, Germany

来源：

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY | 2012年 / 134卷 / 29期

关键词：

ENZYMATIC-SYNTHESIS; DIRECTED EVOLUTION; CRYSTAL-STRUCTURES; NUCLEIC-ACIDS; NUCLEOTIDE INCORPORATION; REVERSIBLE TERMINATOR; LARGE FRAGMENT; LABELED DNA; PROBES; REPLICATION;

D O I：

10.1021/ja3017889

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The capability of DNA polymerases to accept chemically modified nucleotides is of paramount importance for many biotechnological applications. Although these analogues are widely used, the structural basis for the acceptance of the unnatural nucleotide surrogates has been only sparsely explored. Here we present in total six crystal structures of modified 2'-deoxynucleoside-5'-O-triphosphates (dNTPs) carrying modifications at the CS positions of pyrimidines or C7 positions of 7-deazapurines in complex with a DNA polymerase and a primer/template complex. The modified dNTPs are in positions poised for catalysis leading to incorporation. These structural data provide insight into the mechanism of incorporation and acceptance of modified dNTPs. Our results open the door for rational design of modified nucleotides, which should offer great opportunities for future applications.

引用

页码：11840 / 11843

页数：4

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