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Volume 143, Issue 6

The thioredoxin reductase system of mycoplasmas Free

Abstract

Representative species of the Mollicutes possess a thioredoxin reductase system (NTS) composed of a low-molecular-mass thioredoxin (TRX) and NADPH-binding thioredoxin reductase (NTR). The TRXs of and have molecular masses of 11.2 and 12 kDa, respectively, and are stable at 90 °C for 10 min. Both TRXs reacted with monospecific polyclonal antibodies generated against the TRX, but not with anti- TRX antisera. The and NTRs were partially purified and were found to be active with the homologous TRX, but not with the TRX of or The NTS activity had an optimal pH of 6.5-7.5 and was dependent on NADPH as an electron donor, a requirement which could not be fulfilled by NADH. The genes encoding the TRX and NTR ( and ) of were cloned and sequenced. The comparative analysis of the predicted amino acid sequence of trxA showed that the 11.2 kDa protein (102 aa) shared 26-68% sequence similarity with products of other known genes and contained the conserved active site Cys-Gly-Pro-Cys. The predicted amino acid sequence of trxB contained 315 residues with a conserved NADPH binding domain and FAD binding domains I and II. The cysteine dithiol redox active region had isoleucine rather than threonine at the active site, as compared with other NTRs. The high activity of the NTS in mycoplasmas suggests that mycoplasmas may have evolved the NTS to protect themselves from the consequences of their self-generated oxidative challenge.

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Keyword(s): Mollicutes , Mycoplasma capricolum , Mycoplasma pneumoniae , NADP-thioredoxin reductase and thioredoxin
© Society Society for General Microbiology 1997
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1997-06-01
2024-04-20
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References

  1. Aharonowitz, Y.,, Av-Gay, Y.,, Schreiber, R., Cohen, G. 1993; Characterization of a broad-range disulfide reductase from Streptomyces clavuligerus and its possible role in beta-lactam antibiotic biosynthesis. J Bacteriol 175:623–629
     [Google Scholar]
  2. Almagor, M.,, Kahane, I., Yatziv, S. 1984; Role of superoxide anion in host cell injury induced by Mycoplasma pneumoniae infection : a study in normal and trisomy 21 cells. J Clin Invest 73:842–847
     [Google Scholar]
  3. Ansorge, W.,, Sproat, B.,, Stegemann, J.,, Schwager, C., Zenke, M. 1987; Automated DNA sequencing : ultrasensitive detection of fluorescent bands during electrophoresis. Nucleic Acids Res 15:4593–4602
     [Google Scholar]
  4. Ben-Menachem, G.,, Herrmann, R., Rottem, S. 1994; Thioredoxin and thioredoxin reductase in mollicutes. IOM Lett 3:197
     [Google Scholar]
  5. Bork, P.,, Ouzounis, C.,, Casari, G.,, Schneider, R.,, Sander, C.,, Dolan, M.,, Gilbert, W., Gillevet, P. M. 1995; Exploring the Mycoplasma capricolum genome: a minimal cell reveals its physiology.. Mol Microbiol 16:955–967
     [Google Scholar]
  6. Bradford, M. M. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.. Anal Biochem 72:248–254
     [Google Scholar]
  7. Buchanan, B. B.,, Schurmann, P.,, Decottignies, P., Lozano, R. M. 1994; Thioredoxin : a multifunctional regulatory protein with a bright future in technology and medicine.. Arch Biochem Biophys 314:257–260
     [Google Scholar]
  8. Caruthers, M. H. 1982; New methods for synthesizing deoxyoligonucleotides.. Genet Eng 4:1–16
     [Google Scholar]
  9. Chen, N. Y.,, Zhang, J. J., Paulus, H. 1989; Chromosomal location of the Bacillus subtilis aspartokinase II gene and nucleotide sequence of the adjacent genes homologous to uvrC and trx of Escherichia coli.. J Gen Microbiol 135:2931–2940
     [Google Scholar]
  10. Cheng, S.,, Fockler, C.,, Barnes, W. M., Higuchei, R. 1994; Effective amplification of long targets from cloned inserts and human genomic DNA.. Proc Natl Acad Sci USA 91:5695–5699
     [Google Scholar]
  11. Cohen, G.,, Yanko, M.,, Mislovati, M.,, Argaman, A.,, Schreiber, R.,, Av-Gay, Y., Aharonowitz, Y. 1993; Thioredoxin-thioredoxin reductase system of Streptornyces clavuligerus : sequences, expression, and organization of the genes.. J Bacteriol 175:5159–5167
     [Google Scholar]
  12. Cohen, G.,, Argaman, A.,, Schreiber, R.,, Mislovati, M.,, Aharonowitz, Y. 1994; The thioredoxin system of Penicillium chrysogenum and its possible role in penicillin biosynthesis.. J Bacteriol 176:973–984
     [Google Scholar]
  13. Derman, A. I.,, Prinz, W. A.,, Belin, D., Beckwith, J. 1993; Mutations that allow disulfide bond formation in the cytoplasm of Escherichia coli . Science 262:1744–1747
     [Google Scholar]
  14. Fernando, M. R.,, Nanri, H.,, Yoshitake, S.,, Nagata-Kuno, K., Minakami, S. 1992; Thioredoxin regenerates proteins inactivated by oxidative stress in endothelial cells.. Eur J Biochem 209:917–922
     [Google Scholar]
  15. Ferretti, L. S.,, Karnik, S.,, Khorana, H. G.,, Nassal, M., Oprian, D.D. 1992; Total synthesis of a gene for bovine rhodopsin.. Proc Natl Acad Sci USA 83:599–603
     [Google Scholar]
  16. Fleischmann, R. D.and others. 1995; Whole-genome random sequencing and assembly of Haemophilus influenzae, Rd.. Science 269:496–512
     [Google Scholar]
  17. Fraser, C. M. and others. 1995; The minimal gene complement of Mycoplasma genitalium . Science 270:397–403
     [Google Scholar]
  18. Gan, Z. R. 1991; Yeast thioredoxin genes. J Biol Chem 266:2736–2742
     [Google Scholar]
  19. Gleason, F. K., Holmgren, A. 1988; Thioredoxin and related proteins in procaryotes.. FEMS Microbiol Rev 54:271–298
     [Google Scholar]
  20. Himmelreich, R.,, Hilbert, H.,, Plagens, H.,, Pirkl, E.,, Li, B. C, Herrmann, R. 1996; Complete sequence analysis of the genome of the bacterium Mycoplasma pneumoniae.. Nucleic Acids Res 24:4420–4449
     [Google Scholar]
  21. Holmgren, A. 1977; Bovine thioredoxin system : purification of thioredoxin reductase from calf liver and thymus and studies of its function in disulfide reduction.. J Biol Chem 252:4600–4606
     [Google Scholar]
  22. Holmgren, A. 1979a; Reduction of disulfides by thioredoxin. Exceptional reactivity of insulin and suggested functions of thioredoxin in mechanisms of hormone action. J Biol Chem 254:9113–9119
     [Google Scholar]
  23. Holmgren, A. 1979b; Thioredoxin catalyzes the reduction of insulin disulfides by dithiothreitol and dihydrolipoamide. J Biol Chem 254:9627–9632
     [Google Scholar]
  24. Laemmli, U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
     [Google Scholar]
  25. Lim, C.-J.,, Geraghty, D., Fuchs, J. A. 1985; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
     [Google Scholar]
  26. Luthman, M., Holmgren, A. 1982; Rat liver thioredoxin and thioredoxin reductase : purification and characterization.. Bio-Chemistry 21:6628–6633
     [Google Scholar]
  27. Lynch, R.E., Cole, B. c. 1980; Mycoplasma Pneumoniae: a pathogen which manufactures superoxide but lacks superoxide dismutase.. Proc FEBS Symp 62:49–56
     [Google Scholar]
  28. Matthews, J. R.,, Wakasugi, N.,, Virelizier, J. L.,, Yodoi, J., Hay, R.T. 1992; Thioredoxin regulates the DNA binding activity of NFkappa B by reduction of a disulphide bond involving cysteine 62.. Nucleic Acids Res 20:3821–3830
     [Google Scholar]
  29. Pigiet, V. P., Schuster, B. J. 1986; Thioredoxin-catalyzed refolding of disulfide-containing proteins.. Proc Natl Acad Sci USA 83:7634–7647
     [Google Scholar]
  30. Razin, S., Rottem, S. 1976; Mycoplasma membranes. In Biochemical Analysis of Membranes, Edited by A. M. Maddy. London: Chapman and Hall.. pp:3–26
     [Google Scholar]
  31. Russel, M., Model, P. 1988; Sequence of thioredoxin reductase from Escherichia coli. Relationship to other flavoprotein disulfide oxidoreductases.. J Biol Chem 263:9015–9019
     [Google Scholar]
  32. Sambrook, J.,, Fritsch, E. F., Maniatis, T. 1989; Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbour Laboratory.. J Biol Chem 263:9015–9019
     [Google Scholar]
  33. Sanger, F.,, Nicklen, S., Coulson, A. R. 1977; DNA sequencing with chain-terminating inhibitors.. Proc Natl Acad Sci USA 79:5463–5467
     [Google Scholar]
  34. Schenk, H.,, Klein M.,, Erdbrugger, W.,, Droge, W., Schulze, O. K. 1994; Distinct effects of thioredoxin and antioxidants on the activation of transcription factors NF-kappa B and AP-1. Proc Natl Acad Sci USA 91:1672–1676
     [Google Scholar]
  35. Shirazi, I.,, Tarshis, M., Rottem, S. 1995; An arginine-ornithine exchange system in Spiroplasma melliferum cells.. Microbiology 141:2323–2328
     [Google Scholar]
  36. Tryon, V. V., Baseman, J. B. 1992; Pathogenic determinants and mechanisms. In Mycoplasmas : Molecular Biology and Pathogenesis, Edited by J. Maniloff, R. N. McElhaney, L. R. Finch & J . B. Baseman. Washington, DC : American Society for Microbiology.. Microbiology pp:457–471
     [Google Scholar]
  37. Wenzel, R., Herrmann, R. 1988; Physical mapping of the Mycoplasma pneumoniae genome.. Nucleic Acids Res 16:8323–8336
     [Google Scholar]
  38. Wenzel, R., Herrmann, R. 1989; Cloning of the complete Mycoplasma pneumoniae genome.. Nucleic Acids Res 17:7029–7043
     [Google Scholar]
  39. Wollman, E. E.,, D'Auriol, L.,, Rimsky, L.,, Shaw, A.,, Jacquot, J.,, Wingfield, P. P.,, Graber, P., Dessarps, F. 1988; Cloning and expression of a cDNA from human thioredoxin.. J Biol Chem 263:15506–15512
     [Google Scholar]
  40. Wong, J. H.,, Kobrhel, K., Buchanan, B. B. 1995; Thioredoxin and seed proteins.. Methods Enzymol 252:228–240
     [Google Scholar]
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The thioredoxin reductase system of mycoplasmas
Microbiology 143, 1933 (1997); https://doi.org/10.1099/00221287-143-6-1933
/content/journal/micro/10.1099/00221287-143-6-1933
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