Potent anti-tumor effects of an active site mutant of human manganese-superoxide dismutase. Evolutionary conservation of product inhibition

J Biol Chem. 2004 Mar 26;279(13):12769-76. doi: 10.1074/jbc.M310623200. Epub 2003 Dec 18.

Abstract

Mn-SOD serves as the primary cellular defense against oxidative damage by converting superoxide radicals (O(2)(-)) to O(2) and H(2)O(2). A unique characteristic of this mitochondrial anti-oxidant enzyme is the conservation from bacteria to man of a rapidly formed product inhibited state. Using site-directed mutagenesis, we have generated an active site mutant (H30N) of human Mn-SOD, which exhibits significantly reduced product inhibition and increased enzymatic efficiency. Overexpression of the H30N enzyme causes anti-proliferative effects in vitro and anti-tumor effects in vivo. Our results provide a teleological basis for the phylogenetically invariant nature of position His-30 and the evolutionary conservation of product inhibition. These data also provide more direct intracellular evidence for the signaling role associated with H(2)O(2).

MeSH terms

  • Animals
  • Antioxidants / metabolism
  • Binding Sites
  • Blotting, Northern
  • Catalase / metabolism
  • Catalysis
  • Cell Division
  • Cell Line
  • Crystallography, X-Ray
  • DNA, Complementary / metabolism
  • Evolution, Molecular
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Histidine / chemistry
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Immunoblotting
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, SCID
  • Mitochondria / metabolism
  • Models, Chemical
  • Models, Genetic
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Mutation*
  • Oxygen / metabolism*
  • Phylogeny
  • Plasmids / metabolism
  • Retroviridae / genetics
  • Signal Transduction
  • Superoxide Dismutase / genetics*
  • Time Factors
  • Transfection

Substances

  • Antioxidants
  • DNA, Complementary
  • Luminescent Proteins
  • Green Fluorescent Proteins
  • Histidine
  • Hydrogen Peroxide
  • Catalase
  • Superoxide Dismutase
  • Oxygen