Journal
NUCLEIC ACIDS RESEARCH
Volume 47, Issue 5, Pages 2160-2168Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz008
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Funding
- European Research Council under the European Union's Seventh Framework Program (FP7/2007-2013)/ERC [ERC-2012-ADG 20120216/320683]
- FWO Flanders Research Foundation [1247114N]
- Walloon Region (SPW, DGO6, Belgium) [1318159]
- AU-TOBMP2 project [SPW, DGO6] [16100518]
- BBSRC [BB/N01023X/1] Funding Source: UKRI
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Six 1,5-anhydrohexitol uridine triphosphates were synthesized with aromatic substitutions appended via a carboxamide linker to the 5-position of their bases. An improved method for obtaining such 5-substituted hexitol nucleosides and nucleotides is described. The incorporation profile of the nucleotide analogues into a DNA duplex overhang using recently evolved XNA polymerases is compared. Long, mixed HNA sequences featuring the base modifications are generated. The apparent binding affinity of four of the nucleotides to the enzyme, the rate of the chemical step and of product release, plus the specificity constant for the incorporation of these modified nucleotides into a DNA duplex overhang using the HNA polymerase T6G12_I521L are determined via pre-steady-state kinetics. HNA polymers displaying aromatic functional groups could have significant impact on the isolation of stable and high-affinity binders and catalysts, or on the design of nanomaterials.
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