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

    Renders, M. and Dumbre, S. and Abramov, M. and Kestemont, D. and Margamuljana, L. and Largy, E. and Cozens, C. and Vandenameele, J. and Pinheiro, Vitor B. and Toye, D. and Frère, J.-M. and Herdewijn, P. (2019) Kinetic analysis of N-alkylaryl carboxamide hexitol nucleotides as substrates for evolved polymerases. Nucleic Acids Research , ISSN 1362-4962.

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    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.

    Metadata

    Item Type: Article
    Keyword(s) / Subject(s): Genetics
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    SWORD Depositor: Mr Joe Tenant
    Depositing User: Mr Joe Tenant
    Date Deposited: 27 Feb 2019 14:47
    Last Modified: 26 Jul 2019 22:23
    URI: http://eprints.bbk.ac.uk/id/eprint/26137

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