J. Bacteriol., Dec 1996, 7152-7158, Vol 178, No. 24
Copyright © 1996, American Society for Microbiology

Rubrerythrin from Clostridium perfringens: cloning of the gene, purification of the protein, and characterization of its superoxide dismutase function [published erratum appears in J Bacteriol 1997 Mar;179(5):1836]

Y Lehmann, L Meile and M Teuber
Laboratorium fur Lebensmittel-Mikrobiologie, Institut fur Lebensmittelwissenschaften, Eidgenossische Technische Hochschule, ETH- Zentrum, Zurich, Switzerland.

The food-borne pathogen Clostridium perfringens, which is an obligate anaerobe, showed growth under conditions of oxidative stress. In protein extracts we looked for superoxide dismutase (SOD) activities which might scavenge highly toxic superoxide radicals evolving under such stress conditions. Using the classical assay to detect SOD activity on gels after electrophoresis of C. perfringens proteins, we obtained a pattern of three major bands indicating SOD activity. The protein representing the brightest band was purified by three chromatographic steps. On the basis of 20 amino acids determined from the N terminus of the protein, we designed a degenerate oligonucleotide probe to isolate the corresponding gene. We finally sequenced an open reading frame of 195 amino acids (molecular mass, 21,159 Da) with a strong homology to the Desulfovibrio vulgaris rubrerythrin; therefore, we assumed to have cloned a rubrerythrin gene from C. perfringens, and we named it rbr. The C-terminal region of the newly detected rubrerythrin from C. perfringens contains a characteristic non-heme, non-sulfur iron-binding site -Cys-X-X-Cys-(X)12-Cys-X-X-Cys- similar to that found in rubrerythrin from D. vulgaris. In addition, three -Glu-X- X-His- sequences could represent diiron binding domains. We observed SOD activity in extracts of Escherichia coli strains containing the recombinant rbr gene from C. perfringens. A biological function of rubrerythrin as SOD was confirmed with the functional complementation by the rbr gene of an E. coli mutant strain lacking SOD activity. We therefore suppose that rubrerythrin plays a role as a scavenger of oxygen radicals.

This article has been cited by other articles:

• Mukhopadhyay, A., Redding, A. M., Joachimiak, M. P., Arkin, A. P., Borglin, S. E., Dehal, P. S., Chakraborty, R., Geller, J. T., Hazen, T. C., He, Q., Joyner, D. C., Martin, V. J. J., Wall, J. D., Yang, Z. K., Zhou, J., Keasling, J. D. (2007). Cell-Wide Responses to Low-Oxygen Exposure in Desulfovibrio vulgaris Hildenborough. J. Bacteriol. 189: 5996-6010 [Abstract] [Full Text]  
• Hindupur, A., Liu, D., Zhao, Y., Bellamy, H. D., White, M. A., Fox, R. O. (2006). The crystal structure of the E. coli stress protein YciF. Protein Sci. 15: 2605-2611 [Abstract] [Full Text]  
• Kawasaki, S., Watamura, Y., Ono, M., Watanabe, T., Takeda, K., Niimura, Y. (2005). Adaptive Responses to Oxygen Stress in Obligatory Anaerobes Clostridium acetobutylicum and Clostridium aminovalericum. Appl. Environ. Microbiol. 71: 8442-8450 [Abstract] [Full Text]  
• Weinberg, M. V., Jenney, F. E. Jr., Cui, X., Adams, M. W. W. (2004). Rubrerythrin from the Hyperthermophilic Archaeon Pyrococcus furiosus Is a Rubredoxin-Dependent, Iron-Containing Peroxidase. J. Bacteriol. 186: 7888-7895 [Abstract] [Full Text]  
• Jean, D., Briolat, V., Reysset, G. (2004). Oxidative stress response in Clostridium perfringens. Microbiology 150: 1649-1659 [Abstract] [Full Text]  
• Seshadri, R., Myers, G. S. A., Tettelin, H., Eisen, J. A., Heidelberg, J. F., Dodson, R. J., Davidsen, T. M., DeBoy, R. T., Fouts, D. E., Haft, D. H., Selengut, J., Ren, Q., Brinkac, L. M., Madupu, R., Kolonay, J., Durkin, S. A., Daugherty, S. C., Shetty, J., Shvartsbeyn, A., Gebregeorgis, E., Geer, K., Tsegaye, G., Malek, J., Ayodeji, B., Shatsman, S., McLeod, M. P., Smajs, D., Howell, J. K., Pal, S., Amin, A., Vashisth, P., McNeill, T. Z., Xiang, Q., Sodergren, E., Baca, E., Weinstock, G. M., Norris, S. J., Fraser, C. M., Paulsen, I. T. (2004). Comparison of the genome of the oral pathogen Treponema denticola with other spirochete genomes. Proc. Natl. Acad. Sci. USA 101: 5646-5651 [Abstract] [Full Text]  
• Fournier, M., Zhang, Y., Wildschut, J. D., Dolla, A., Voordouw, J. K., Schriemer, D. C., Voordouw, G. (2003). Function of Oxygen Resistance Proteins in the Anaerobic, Sulfate-Reducing Bacterium Desulfovibrio vulgaris Hildenborough. J. Bacteriol. 185: 71-79 [Abstract] [Full Text]  
• Briolat, V., Reysset, G. (2002). Identification of the Clostridium perfringens Genes Involved in the Adaptive Response to Oxidative Stress. J. Bacteriol. 184: 2333-2343 [Abstract] [Full Text]  
• Meile, L., Rohr, L. M., Geissmann, T. A., Herensperger, M., Teuber, M. (2001). Characterization of the D-Xylulose 5-Phosphate/D-Fructose 6-Phosphate Phosphoketolase Gene (xfp) from Bifidobacterium lactis. J. Bacteriol. 183: 2929-2936 [Abstract] [Full Text]  
• Das, A., Coulter, E. D., Kurtz, D. M. Jr., Ljungdahl, L. G. (2001). Five-Gene Cluster in Clostridium thermoaceticum Consisting of Two Divergent Operons Encoding Rubredoxin Oxidoreductase- Rubredoxin and Rubrerythrin-Type A Flavoprotein- High-Molecular-Weight Rubredoxin. J. Bacteriol. 183: 1560-1567 [Abstract] [Full Text]  
• Lumppio, H. L., Shenvi, N. V., Summers, A. O., Voordouw, G., Kurtz, D. M. Jr. (2001). Rubrerythrin and Rubredoxin Oxidoreductase in Desulfovibrio vulgaris: a Novel Oxidative Stress Protection System. J. Bacteriol. 183: 101-108 [Abstract] [Full Text]  
• Ratnayake, D. B., Wai, S. N., Shi, Y., Amako, K., Nakayama, H., Nakayama, K. (2000). Ferritin from the obligate anaerobe Porphyromonas gingivalis: purification, gene cloning and mutant studies. Microbiology 146: 1119-1127 [Abstract] [Full Text]  
• GUTTERIDGE, J. M.C., HALLIWELL, B. (2000). Free Radicals and Antioxidants in the Year 2000: A Historical Look to the Future. Ann. N. Y. Acad. Sci. 899: 136-147 [Abstract] [Full Text]  
• Geissmann, T. A., Teuber, M., Meile, L. (1999). Transcriptional Analysis of the Rubrerythrin and Superoxide Dismutase Genes of Clostridium perfringens. J. Bacteriol. 181: 7136-7139 [Abstract] [Full Text]  
• Jenney Jr., F. E., Verhagen, M. F., Cui, X., Adams, M. W. (1999). Anaerobic Microbes: Oxygen Detoxification Without Superoxide Dismutase. Science 286: 306-309 [Abstract] [Full Text]  
• Voordouw, J. K., Voordouw, G. (1998). Deletion of the rbo Gene Increases the Oxygen Sensitivity of the Sulfate-Reducing Bacterium Desulfovibrio vulgaris Hildenborough. Appl. Environ. Microbiol. 64: 2882-2887 [Abstract] [Full Text]  
• Liochev, S. I., Fridovich, I. (1997). A Mechanism for Complementation of the sodA sodB Defect in Escherichia coli by Overproduction of the rbo Gene Product (Desulfoferrodoxin) from Desulfoarculus baarsii. J. Biol. Chem. 272: 25573-25575 [Abstract] [Full Text]