Kinetic analysis of N-alkylaryl carboxamide hexitol nucleotides as substrates for evolved polymerases
| Autor: | Shrinivas G. Dumbre, Dominique Toye, Piet Herdewijn, Christopher Cozens, Vitor B. Pinheiro, Julie Vandenameele, Jean-Marie Frère, Marleen Renders, Mikhail Abramov, Donaat Kestemont, Eric Largy, Lia Margamuljana |
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| Rok vydání: | 2019 |
| Předmět: |
Specificity constant
Stereochemistry medicine.drug_class Carboxamide DNA-Directed DNA Polymerase Biology bcs Protein Engineering Substrate Specificity 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Sugar Alcohols Chemical Biology and Nucleic Acid Chemistry Genetics medicine Nucleotide A-DNA Polymerase 030304 developmental biology chemistry.chemical_classification 0303 health sciences Nucleotides Kinetics chemistry biology.protein Linker Uracil nucleotide 030217 neurology & neurosurgery DNA |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 |
| Popis: | 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. ispartof: NUCLEIC ACIDS RESEARCH vol:47 issue:5 pages:2160-2168 ispartof: location:England status: published |
| Databáze: | OpenAIRE |
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