Abstract
The crenarchaeon Pyrobaculum aerophilum is with an optimalgrowth temperature of 100 °C one of the most thermophilic organisms knownto possess an aerobic respiratory chain. The analysis of DNA sequences fromthe Pyrobaculum genome project lead to the identification of an openreading frame potentially coding for a Rieske iron-sulfur protein. Thecomplete gene (named parR) was cloned and sequenced. The deducedamino acid sequence displays unusual amino acid exchanges and a so farunknown sequence insertion. The N-terminus shows similarities to bacterialsignal sequences. Several forms of the gene were expressed in E.coli in order to verify the classification as a Rieske protein and tofacilitate biophysical studies. Soluble, thermo-stable proteins withcorrectly inserted iron-sulfur clusters were expressed from two versions ofthe gene. The Δ1–23 truncated holo-protein is redox active. Itdisplays the typical spectroscopic properties of a Rieske protein. The redoxpotential was determined to be +215 mV at pH 6.5 and is pH dependentabove pH 7.5 revealing the influence of two protonation equilibria with pKavalues of 8.1 and 9.8. Phylogenetic analysis demonstrates that the parRprotein clusters together with the two other available archaeal Rieskesequences from Sulfolobus on a separate branch of the phylogenetictree apart from the proteins from thermophilic bacteria like Aquifexand Thermus.
This is a preview of subscription content, log in via an institution to check access.
REFERENCES
Anemüller, S., Schmidt, C. L., Schäfer, G., Bill, E., Trautwein, A. X., and Teixeira, M. (1994). Biochem. Biophys. Res. Commun. 202, 252–257.
Burggraf, S., Huber, H., and Stetter, K. O. (1997). Int. J. Syst. Bacteriol. 47, 657–660.
Carrell, C. J., Zhang, H., Cramer, W. A., and Smith, J. L. (1997). Structure 5, 1613–1625.
Castresana, J., Lübben, M., and Saraste, M. (1995). J. Mol. Biol. 250, 202–210.
Clark, M. W. (1960). Oxidation-reduction-potentials of organic systems, Williams and Wilkins, Baltimore.
Denke, E., Merbitz-Zahradnik, T., Hatzfeld, O. M., Snyder, C. H., Link, T. A., and Trumpower, B. L. (1998). J. Biol. Chem. 273, 9085–9093.
Engelman, D. M., Steitz, T. A., and Goldman, A. (1986). Ann. Rev. Biophys. Biophys. Chem. 15, 321–353.
Fitz-Gibbon, S., Choi, A. J., Miller, J. H., Stetter, K. O., Simon, M. I., Swanson, R., Kim, U.-J. (1997). Extremophiles 1, 36–51.
Gambacorta, A., Trincone, A., Nicolaus, B., Lama, L., and de Rosa, M. (1994). System. Appl. Microbiol. 16, 518–527.
Guex, N., and Peitsch, M. C. (1997). Electrophoresis 18, 2714–2723.
Iwasaki, T., Imai, T., Urushiyama, A., and Oshima, T. (1996). J. Biol. Chem. 271, 27659–27663.
Iwasaki, T., Isogai, Y., Iizuka, T., and Oshima, T. (1995). J. Bacteriol. 177, 2576–2582.
Iwata, S., Lee, J. W., Okada, K., Lee, J. K., Iwata, M., Rasmussen, B., Link, T. A., Ramaswamy, S., Jap, B. K. (1998). Science 281, 64–71.
Iwata, S., Saynovits, M., Link, T. A., and Michel, H. (1996). Structure 4, 567–579.
Kuila, D., and Fee, J. A. (1986). J. Biol. Chem. 261, 2768–2771.
Link, T. A., Hagen, W. R., Pierik, A. J., Assmann, C., and von Jagow, G. (1992). Eur. J. Biochem. 208, 685–691.
Lübben, M. (1995). Biochim. Biophys. Acta 1229, 1–22.
Lübben, M., Arnaud, S., Castresana, J., Warne, A., Albracht, S. P. J., and Saraste, M. (1994). Eur. J. Biochem. 224, 151–159.
Mason, J. R., and Cammack, R. (1992). Annu. Rev. Microbiol. 46, 277–305.
Nozawa, T., Trost, J. T., Fukada, T., Hatano, M., McManus, J. D., and Blankenship, R. E. (1987). Biochim. Biophys. Acta 894, 468–476.
Riedel, A., Kellner, E., Grodzitzki, D., Liebl, U., Hauska, G., Müller, A., Rutherford, A. W., and Nietschke, W. (1993). Biochim. Biophys. Acta 1183, 263–268.
Rost, B., and Sander, C. (1994). Proteins 19, 55–77.
Rost, B., and Sander, C. (1993). J. Mol. Biol. 232, 584–599.
Schmidt, C. L., Anemüller, S., Schäfer, G. (1996). FEBS Lett. 388, 43–46.
Schmidt, C. L., Anemüller, S., Teixeira, M., Schäfer, G. (1995). FEBS Lett. 359, 239–243.
Schmidt, C. L., Hatzfeld, O. M., Petersen, A., Link, T. A., and Schäfer, G. (1997). Biochem. Biophys. Res. Commun. 234, 283–287.
Schoepp, B., Brugna, M., Lebrun, E., and Nitschke, W. (1999). Adv. Inorg. Chem., in press.
Schröter, T., Hatzfeld, O. M., Gemeinhardt, S., Korn, M., Friedrich, T., Ludwig, B., Link, T. A. (1998). Eur. J. Biochem. 255, 100–106.
Trumpower, B. L. (1981). Biochim. Biophys. Acta 639, 129–155.
Van Doren, S. R., Yun, C.-H., Crofts, A. R., and Gennis, R. B. (1993). Biochemistry 32, 628–636.
Völkl, P., Huber, R., Drobner, E., Rachel, R., Burggraf, S., Trincone, A., and Stetter, K. O. (1993). Appl. Environ. Microbiol. 59, 2918–2926.
Zhang, H., Carrell, C. J., Huang, D., Sled, V., Ohnishi, T., Smith, J. L., and Cramer, W. A. (1996). J. Biol. Chem. 271, 31360–31366.
Zhang, Z., Huang, L., Shulmeister, V. M., Chi, Y. I., Kim, K. K., Hung, L. W., Crofts, A. R., Berry, E. A., and Kim, S. H. (1998). Nature 392, 677–684.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Henninger, T., Anemüller, S., Fitz-Gibbon, S. et al. A Novel Rieske Iron-Sulfur Protein from the Hyperthermophilic Crenarchaeon Pyrobaculum aerophilum: Sequencing of the Gene, Expression in E. coli and Characterization of the Protein. J Bioenerg Biomembr 31, 119–128 (1999). https://doi.org/10.1023/A:1005447710894
Issue Date:
DOI: https://doi.org/10.1023/A:1005447710894