Spectroelectrochemical insights into structural and redox properties of immobilized endonuclease III and its catalytically inactive mutant

Elin Moe, Filipe Rollo, Célia M. Silveira, Murat Sezer, Peter Hildebrandt, Smilja Todorovic

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Endonuclease III is a Fe-S containing bifunctional DNA glycosylase which is involved in the repair of oxidation damaged DNA. Here we employ surface enhanced IR spectroelectrochemistry and electrochemistry to study the enzyme from the highly radiation- and desiccation-resistant bacterium Deinococcus radiodurans (DrEndoIII2). The experiments are designed to shed more light onto specific parameters that are currently proposed to govern damage search and recognition by endonucleases III. We demonstrate that electrostatic interactions required for the redox activation of DrEndoIII2 may result in high electric fields that alter its structural and thermodynamic properties. Analysis of inactive DrEndoIII2 (K132A/D150A double mutant) interacting with undamaged DNA, and the active enzyme interacting with damaged DNA also indicate that the electron transfer is modulated by subtle differences in the protein-DNA complex.

Original languageEnglish
Pages (from-to)149-154
Number of pages6
JournalSpectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy
Volume188
DOIs
Publication statusPublished - 5 Jan 2018

Keywords

  • 4Fe-4S cluster
  • Cyclic voltammetry
  • DNA repair
  • Endonuclease III
  • Resonance Raman spectroscopy
  • Surface enhanced IR absorption

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