Manganese-containing superoxide dismutase and its gene from Candida albicans

Mitochondrial manganese-containing superoxide dismutase was purified around 112-fold with an overall yield of 1.1% to apparent electrophoretic homogen eity from the dimorphic pathogenic fungus, Candida albicans. The molecular mass of the native enzyme was 106 kDa and the enzyme was composed of four i dentical subunits with a molecular mass of 26 kDa. The enzyme was not sensi tive to either cyanide or hydrogen peroxide. The N-terminal amino acid sequ ence alignments (up to the 18th residue) showed that the enzyme has high si milarity 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 a nalysis of the sod2 predicted a manganese-containing superoxide dismutase t hat contains 234 amino acid residues with a molecular mass of 26 173 Da, an d displayed 57% sequence identity to the homologue of Saccharomyces cerevis iae. The deduced N-terminal 34 amino acid residues may serve as a signal pe ptide for mitochondrial translocation. Several regulatory elements such as stress responsive element and haem activator protein 2/3/4/5 complex bindin g sites were identified in the promoter region of sod2. Northern analysis w ith a probe derived from the cloned sod2 revealed a 0.94-kb band, which cor responds approximately to the expected size of mRNA deduced from sod2. (C) 1999 Elsevier Science B.V. All rights reserved.