Nonenzymatic cleavage of proteins by reactive oxygen species generated by dithiothreitol and iron

J Biol Chem. 1985 Dec 15;260(29):15394-7.

Abstract

Many enzymes, represented by yeast glutamine synthetase, are inactivated and degraded in the presence of dithiothreitol (DTT), oxygen, and catalytic amounts of iron salts. The roles of DTT and iron can be replaced by ascorbate and copper, respectively. Experimental data suggest that reactive oxygen species, likely hydroxyl radicals, are generated locally around irons bound at specific sites on enzymes, and these species are responsible for the inactivation and degradation. Since many biochemicals are contaminated with metal salts in quantities sufficient for some hydroxyl radical formation to occur, the possibility of oxidative modification and degradation should be considered when an enzyme is exposed to DTT.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Cattle
  • Chromatography, High Pressure Liquid
  • Dithiothreitol / pharmacology*
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli
  • Glutamate-Ammonia Ligase / metabolism*
  • Iron / pharmacology*
  • Models, Chemical
  • Oxidation-Reduction
  • Oxygen / metabolism*
  • Rabbits
  • Saccharomyces cerevisiae / enzymology
  • Time Factors

Substances

  • Amino Acids
  • Iron
  • Glutamate-Ammonia Ligase
  • Oxygen
  • Dithiothreitol