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Kinetic analysis of N-alkylaryl carboxamide hexitol nucleotides as substrates for evolved polymerases.

Abstract：

: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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Renders M,Dumbre S,Abramov M,Kestemont D,Margamuljana L,Largy E,Cozens C,Vandenameele J,Pinheiro VB,Toye D,Frère JM,Herdewijn P

doi

10.1093/nar/gkz008

subject

Has Abstract

pub_date

2019-03-18 00:00:00

pages

2160-2168

issue

5

eissn

0305-1048

issn

1362-4962

pii

5304328

journal_volume

47

pub_type

杂志文章

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