Copper- and zinc-containing superoxide dismutase and its gene from Candida albicans

Biochim Biophys Acta. 1999 Apr 19;1427(2):245-55. doi: 10.1016/s0304-4165(99)00020-3.

Abstract

Cytosolic copper- and zinc-containing superoxide dismutase was purified 136-fold with an overall yield of 2.5% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, Candida albicans. The molecular mass of the native enzyme was 39.4 kDa and the enzyme was composed of two identical subunits with a molecular mass of 19.6 kDa. The enzyme was stable in the range of pH 4.0-9.0 and up to 55 degrees C. The ultraviolet-visible absorption spectrum of the enzyme showed the absorption band of copper- and zinc-containing superoxide dismutase at 660 nm. The atomic absorption analysis revealed that the enzyme contained 0.87 g-atom of copper and 0.79 g-atom of zinc per mole of subunit. The N-terminal amino acid sequence alignments up to the 40th residue showed that copper- and zinc-containing superoxide dismutase from C. albicans has high similarity to other eukaryotic copper- and zinc-containing superoxide dismutases. The sod1 encoding copper- and zinc-containing superoxide dismutase has been cloned using a polymerase chain reaction fragment as a probe. Sequence analysis of the sod1 predicted a copper- and zinc-containing superoxide dismutase that contains 154 amino acids with a molecular mass of 16143 Da and displayed 79%, 69%, and 57% sequence identity to the homologues of Neurospora crassa, Saccharomyces cerevisiae, and bovine, respectively. The cloned sod1 contained an intron of 245 nucleotides, which was verified by reverse transcription-polymerase chain reaction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Candida albicans / enzymology
  • Candida albicans / genetics*
  • Cloning, Molecular
  • Copper / analysis
  • Cytosol / enzymology
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Molecular Sequence Data
  • Molecular Weight
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Spectrophotometry
  • Superoxide Dismutase / chemistry
  • Superoxide Dismutase / isolation & purification*
  • Superoxide Dismutase-1
  • Temperature
  • Zinc / analysis

Substances

  • Copper
  • Superoxide Dismutase
  • Superoxide Dismutase-1
  • Zinc

Associated data

  • GENBANK/AF046872