Abstract
A manganese-containing superoxide dismutase (EC 1.15.1.1) was purified to homogeneity from a higher plant for the first time. The enzyme was isolated fromPisum sativum leaf extracts by thermal fractionation, ammonium sulfate salting out, ion-exchange and gel-filtration column chromatography, and preparative polyacrylamide gel electrophoresis. Pure manganese superoxide dismutase had a specific activity of about 3,000 U mg-1 and was purified 215-fold, with a yield of 1.2 mg enzyme per kg whole leaf. The manganese superoxide dismutase had a molecular weight of 94,000 and contained one g-atom of Mn per mol of enzyme. No iron and copper were detected. Activity reconstitution experiments with the pure enzyme ruled out the possibility of a manganese loss during the purification procedure. The stability of manganese superoxide dismutase at-20°C, 4°C, 25°C, 50°C, and 60°C was studied, and the enzyme was found more labile at high temperatures than bacterial manganese superoxide dismutases and iron superoxide dismutases from an algal and bacterial origin.
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Abbreviations
- NBT:
-
nitro blue tetrazolium
- SOD:
-
superoxide dismutase (EC 1.15.1.1)
References
Arron, G., Henry, L., Palmer, J.M., Hall, D.O. (1976) Superoxide dismutases in mitochondria fromHelianthus tuberosus andNeurospora crassa. Biochem. Soc. Trans.4, 618–620
Asada, K., Kanematsu, S., Uchida, K. (1977) Superoxide dismutases in photosynthetic organisms. Absence of the cuprozinc enzyme in eukaryotic algae. Arch. Biochem. Biophys.179, 243–256
Asada, K., Kanematsu, S., Takahashi, M., Kono, Y. (1976) Superoxide dismutases in photosynthetic organisms. In: Iron and copper proteins, pp. 551–564, Yasunobu, K.T., Mower, H.F., Hayaishi, O., eds. Plenum, New York
Asada, K., Urano, M., Takahashi, M. (1973) Subcellular location of superoxide dismutase in spinach leaves and preparation and properties of crystalline spinach superoxide dismutase. Eur. J. Biochem.36, 257–266
Beauchamp, C.O., Fridovich, I. (1973) Isozymes of superoxide dismutase from wheat germ. Biochim. Biophys. Acta317, 50–64
Beauchamp, C.O., Fridovich, I. (1971) Superoxide dismutase. Improved assays and an assay applicable to acrylamide gels. Anal. Biochem.44, 276–287
Davis, B.J. (1964) Disc gel electrophoresis. Ann. N. Y. Acad. Sci.121, 404–427
Del Río, L.A., Sevilla, F., Gómez, M., Yáñez, J., López-Gorgé, J. (1978a) Superoxide dismutase. An enzyme system for the study of micronutrient interactions in plants. Planta140, 221–225
Del Río, L.A., Gómez, M., Yáñez, J., Leal, A., López-Gorgé, J. (1978b) Iron deficiency in pea plants. Effect on catalase, peroxidase, chlorophyll and proteins of leaves. Plant Soil49, 343–353
Foyer, C.H., Hall, D.O. (1979) A rapid procedure for the preparation of light harvesting chlorophyll a/b protein complex. FEBS Lett.101, 324–328
Giannopolitis, C.N., Ries, S.K. (1977) Superoxide dismutase. I. Occurrence in higher plants. Plant Physiol.59, 309–314
Hakamata, K., Kanematsu, S., Asada, K. (1978) Superoxide dismutase in tea leaf. Study of Tea54, 6–10
Harris, J.I. (1977) Superoxide dismutase fromBacillus stearothermophilus. In: Superoxide and superoxide dismutases, pp. 151–157, Michelson, A.M., McCord, J.M., Fridovich, I., eds. Academic Press, New York
Hedrick, J.L., Smith, A.J. (1968) Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch. Biochem. Biophys.126, 155–164
Ichihara, K., Kusunose, E., Kusunose, M., Mori, T. (1977) Superoxide dismutase fromMycobacterium lepraemurium. J. Biochem.81, 1427–1433
Jackson, C., Dench, J., Moore, A.L., Halliwell, B., Foyer, C.H., Hall, D.O. (1978) Subcellular localisation and identification of superoxide dismutase in the leaves of higher plants. Eur. J. Biochem.91, 339–344
Kanematsu, S., Asada, K. (1978) Superoxide dismutase from an anaerobic photosynthetic bacterium,Chromatium vinosum. Arch. Biochem. Biophys.185, 473–482
Kanematsu, S., Asada, K. (1979) Ferric and manganic superoxide dismutases inEuglena gracilis. Arch. Biochem. Biophys.195, 535–545
Keele, B.B., McCord, J.M., Fridovich, I. (1970) Superoxide dismutase fromEscherichia coli B: a new manganese-containing enzyme. J. Biol. Chem.245, 6176–6181
Kono, Y., Takahashi, M., Asada, K. (1979) Superoxide dismutases from kidney bean leaves. Plant Cell Physiol.20, 1229–1235
Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem.193, 265–275
Lumsden, J., Hall, D.O. (1974) Soluble and membrane-bound superoxide dismutases in a blue-green alga (Spirulina) and spinach. Biochem. Biophys. Res. Commun.58, 35–41
Lumsden, J., Hall, D.O. (1975) Chloroplast manganese and superoxide. Biochem. Biophys. Res. Commun.64, 595–602
Lumsden, J., Cammack, R., Hall, D.O. (1976) Purification and physicochemical properties of superoxide dismutase from two photosynthetic microorganisms. Biochim. Biophys. Acta438, 380–392
McCord, J.M., Fridovich, I. (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J. Biol. Chem.244, 6049–6055
Misra, H.P., Fridovich, I. (1977) Purification and properties of superoxide dismutase from a red alga,Porphyridium cruentum. J. Biol. Chem.252, 6421–6423
Ose, D.E., Fridovich, I. (1976) Superoxide dismutase. Reversible removal of manganese and its substitution by cobalt, nickel, or zinc. J. Biol. Chem.251, 1217–1218
Potty, V.H. (1969) Determination of proteins in the presence of phenols and pectins. Anal. Biochem.29, 535–539
Puget, K., Michelson, A.M. (1974) Iron-containing superoxide dismutases from luminous bacteria. Biochimie56, 1255–1267
Salin, M.L., Day, E.D., Jr., Crapo, J.D. (1978) Isolation and characterization of a manganese-containing superoxide dismutase from rat liver. Arch. Biochem. Biophys.187, 223–228
Salin, M.L. (1980) Isolation and characterisation of an iron-containing superoxide dismutase fromBrassica campestris. In: The significance of superoxide and superoxide dismutase, vol. I, Bannister, J.V., Hill, H.A.O., eds. Elsevier, Amsterdam
Sato, S., Harris, J.I. (1977) Superoxide dismutase fromThermus aquaticus. Isolation and characterization of manganese and apo enzymes. Eur. J. Biochem.73, 373–381
Sawada, Y., Ohyama, T., Yamazaki, I. (1972) Preparation and physicochemical properties of green pea superoxide dismutase. Biochim. Biophys. Acta268, 305–312
Vance, P.T., Keele, B.B., Jr., Rajagopalan, K.V. (1972) Superoxide dismutase fromStreptococcus mutans. Isolation and characterization of two forms of the enzyme. J. Biol. Chem.247, 4782–4786
Von Kameke, E., Wegmann, K. (1977) Isolation of manganese-containing subchloroplast fractions fromDunaliella. Plant Sci. Lett.8, 1–5
Weisiger, R.A., Fridovich, I. (1973) Superoxide dismutase. Organelle specificity. J. Biol. Chem.248, 3582–3592
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Sevilla, F., López-Gorgé, J., Gómez, M. et al. Manganese superoxide dismutase from a higher plant. Planta 150, 153–157 (1980). https://doi.org/10.1007/BF00582359
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DOI: https://doi.org/10.1007/BF00582359