Manganese-containing superoxide dismutase and its gene from Candida albicans

• Published in: Biochimica et biophysica acta Vol. 1426; no. 3; pp. 409 - 419
• Main Authors: Rhie, Gi-eun, Hwang, Cheol-Sang, Brady, MartinJ, Kim, Seong-Tae, Kim, Yeon-Ran, Huh, Won-Ki, Baek, Yong-Un, Lee, Byung-Hoon, Lee, Jung-Sin, Kang, Sa-Ouk
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 02.02.1999
• Subjects: Amino Acid Sequence; Base Sequence; Candida albicans; Candida albicans - enzymology; Candida albicans - genetics; Cloning, Molecular; genbank/af031478; genes; Genes, Fungal; Manganese - analysis; Manganese-containing superoxide dismutase; Molecular Sequence Data; Molecular Weight; Nucleotide sequence; nucleotide sequences; Open reading frame; Sequence Alignment; Sod2; superoxide dismutase; Superoxide Dismutase - chemistry; Superoxide Dismutase - genetics; Superoxide Dismutase - isolation & purification; Candida albicans; Manganese-containing superoxide dismutase; Nucleotide sequence; Sod2; Open reading frame
• ISSN: 0304-4165; 0006-3002; 1872-8006; 1878-2434
• DOI: 10.1016/S0304-4165(98)00161-5

Mitochondrial manganese-containing superoxide dismutase was purified around 112-fold with an overall yield of 1.1% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, Candida albicans. The molecular mass of the native enzyme was 106 kDa and the enzyme was composed of four identical subunits with a molecular mass of 26 kDa. The enzyme was not sensitive to either cyanide or hydrogen peroxide. The N-terminal amino acid sequence alignments (up to the 18th residue) showed that the enzyme has high similarity to the other eukaryotic manganese-containing superoxide dismutases. The gene sod2 encoding manganese-containing superoxide dismutase has been cloned using a product obtained from polymerase chain reaction. Sequence analysis of the sod2 predicted a manganese-containing superoxide dismutase that contains 234 amino acid residues with a molecular mass of 26 173 Da, and displayed 57% sequence identity to the homologue of Saccharomyces cerevisiae. The deduced N-terminal 34 amino acid residues may serve as a signal peptide for mitochondrial translocation. Several regulatory elements such as stress responsive element and haem activator protein 2/3/4/5 complex binding sites were identified in the promoter region of sod2. Northern analysis with a probe derived from the cloned sod2 revealed a 0.94-kb band, which corresponds approximately to the expected size of mRNA deduced from sod2.