5-Substituted Pyrimidine and 7-Substituted 7-Deazapurine dNTPs as Substrates for DNA Polymerases in Competitive Primer Extension in the Presence of Natural dNTPs

0469259 - ÚOCHB 2017 RIV US eng J - Journal Article
Cahová, Hana - Panattoni, Alessandro - Kielkowski, Pavel - Fanfrlík, Jindřich - Hocek, Michal
5-Substituted Pyrimidine and 7-Substituted 7-Deazapurine dNTPs as Substrates for DNA Polymerases in Competitive Primer Extension in the Presence of Natural dNTPs.
ACS Chemical Biology. Roč. 11, č. 11 (2016), s. 3165-3171. ISSN 1554-8929. E-ISSN 1554-8937
R&D Projects: GA ČR GA14-04289S
EU Projects: European Commission(XE) 642023 - ClickGene
Institutional support: RVO:61388963
Keywords : enzymatic synthesis * 2'-deoxyribonucleoside triphosphates * restriction endonucleases
OECD category: Biochemistry and molecular biology
Impact factor: 4.995, year: 2016
Method of publishing: Open access
http://pubs.acs.org/doi/full/10.1021/acschembio.6b00714

A complete series of 5-substituted uracil or cytosine, as well as 7-substituted 7-deazaadenine and 7-deazaguanine 2'-deoxyribonucleoside triphosphates (dNTPs) bearing substituents of increasing bulkiness (H, Me, vinyl, ethynyl, and phenyl) were systematically studied in competitive primer extension in the presence of their natural counterparts (nonmodified dNTPs), and their kinetic data were determined. The results show that modified dNTPs bearing, pi-electron containing substituents (vinyl, ethynyl, Ph) are typically excellent substrates for DNA polymerases comparable to or better than natural dNTPs. The kinetic studies revealed that these modified dNTPs have higher affinity to the active site of the enzyme primer template complex, and the calculations (semiempirical quantum mechanical scoring function) suggest that it is due to the cation-pi interaction of the modified dNTP with Arg629 in the active site of Bst DNA polymerase.
Permanent Link: http://hdl.handle.net/11104/0267059