Superoxide dismutase from Mycobacterium tuberculosis.
1. A superoxide dismutase [EC 1.15.1.1] was purified about 275-fold with a yield of 34% from Mycobacterium tuberculosis, strain H37Ra (attenuated strain), grown on a Sauton medium for two months. The purified enzyme was homogeneous as judged by polyacrylamide gel electrophoresis, and by analytical ultracentrifugation and sedimentation equilibrium studies. 2. The molecular weight of the enzyme was estimated to be approximately 88,000 by sedimentation equilibrium analysis. Since the molecular weight of the subunit was 21,000 as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, the enzyme appears to be composed of four subunits of equal size. 3. Electron spin resonance (ESR) spectra showed that the enzyme contained ferric iron, and metal analysis showed that the enzyme contained ferric iron, and metal analysis showed that approximately 3.7 atoms of iron were present per mole of the enzyme, indicating the occurrence of 1 atom of iron per subunit. 4. The amino acid composition was apparently similar to those of the iron-containing superoxide dismutases from Escherichia coli, luminous bacteria, Pseudomonas ovalis, and blue-green alga. 5. Antibodies against the enzyme were raised in rabbits and immunological studies were performed. The enzyme from M. tuberculosis, strain H37Rv (virulent strain), was found to have antigenic structures identical with those of the H37Ra enzyme. On the other hand, the manganese-containing superoxide dismutases from other species of mycobacteria, i.e., Mycobacterium species, strain Takeo, M. phlei and M. lepraemurium, showed only partial immunological identity with the H37Ra enzyme. 6. During the growth of M. tuberculosis, strain H37Ra, the enzyme was found to be secreted into the culture medium.
D000596:Amino Acids / Q000032:analysis; D000918:Antibody Specificity; D007501:Iron; D046911:Macromolecular Substances; D008970:Molecular Weight; D009161:Mycobacterium / Q000201:enzymology; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D011485:Protein Binding; D013482:Superoxide Dismutase / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature
D000596:Amino Acids; D046911:Macromolecular Substances; D007501:Iron; D013482:Superoxide Dismutase