Insights into substrate recognition by the Escherichia coli Orf135 protein through its solution structure

Biochem Biophys Res Commun. 2012 Apr 6;420(2):263-8. doi: 10.1016/j.bbrc.2012.02.146. Epub 2012 Mar 5.

Abstract

Escherichia coli Orf135 hydrolyzes oxidatively damaged nucleotides such as 2-hydroxy-dATP, 8-oxo-dGTP and 5-hydroxy-CTP, in addition to 5-methyl-dCTP, dCTP and CTP. Nucleotide pool sanitization by Orf135 is important since nucleotides are continually subjected to potential damage by reactive oxygen species produced during respiration. Orf135 is a member of the Nudix family of proteins which hydrolyze nucleoside diphosphate derivatives. Nudix hydrolases are characterized by the presence of a conserved motif, even though they recognize various substrates and possess a variety of substrate binding pockets. We investigated the tertiary structure of Orf135 and its interaction with a 2-hydroxy-dATP analog using NMR. We report on the solution structure of Orf135, which should contribute towards a structural understanding of Orf135 and its interaction with substrates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / chemistry
  • Amino Acid Motifs
  • Binding Sites
  • Escherichia coli / enzymology*
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Conformation
  • Pyrophosphatases / chemistry*
  • Substrate Specificity

Substances

  • 2-hydroxyadenosine-5'-triphosphate
  • Adenosine Triphosphate
  • Pyrophosphatases
  • nudix hydrolase Orf135