1887

Abstract

A mitochondrion-like organelle (MLO) was isolated from isotonic homogenates of . The organelle sedimented at 5000  for 10 min, and had an isopycnic density in sucrose of 1.2 g ml. Biochemical characterization enabled the demonstration of several key enzymes that allowed the construction of a metabolic pathway consisting of an incomplete Krebs cycle linked to the oxygen-sensitive enzymes pyruvate : NADP oxidoreductase (PNO), acetate : succinate CoA transferase (ASCT) and succinate thiokinase (STK), which cumulatively are responsible for recycling CoA and generating ATP. The organelle differs from typical aerobic mitochondria in possessing an oxygen-sensitive PNO that can use FAD or FMN as electron acceptor but is inactive with NAD, ferredoxin or ferredoxin. A gene with 77 % sequence similarity to the PNO mitochondrion precursor cluster from sp[Q941N5] was identified in the genome database. A second cluster with 56 % sequence similarity to the pyruvate : ferredoxin oxidoreductase (PFOR) from was also identified, which is in agreement with the concept that the PNO gene arose through the fusion of a eubacterial gene for PFOR with the gene for NADPH : cytochrome p450 reductase. Hydrogenase activity was not detected under the conditions used in this study. The oranelle therefore demonstrates significant biochemical differences from traditional mitochondria and hydrogenosomes, but possesses features of both. Based upon the results of this study, the organelle falls into the category of a MLO.

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2008-09-01
2019-10-19
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References

  1. Akhmanova, A., Voncken, F., van Alen, T., van Hoek, A., Boxma, B., Vogels, G., Veenhuis, M. & Hackstein, J. H. ( 1998; ). A hydrogenosome with a genome. Nature 396, 527–528.[CrossRef]
    [Google Scholar]
  2. Borooah, J., Leaback, D. H. & Walker, P. G. ( 1961; ). Studies on glucosaminidase. 2. Substrates for N-acetyl-β-glucosaminidase. Biochem J 78, 106–110.
    [Google Scholar]
  3. Boyer, P. D. ( 1962; ). Pyruvate kinase. In The Enzymes, pp. 95–113. Edited by P. D. Boyer, K. H. Lardy & K. Myrback. New York: Academic Press.
  4. Burri, L., Williams, B. A., Bursac, D., Lithgow, T. & Keeling, P. J. ( 2006; ). Microsporidia mitosomes retain elements of the general mitochondrial targeting system. Proc Natl Acad Sci U S A 103, 15916–15920.[CrossRef]
    [Google Scholar]
  5. Colombo, S., Grisa, M., Tortora, P. & Vanoni, M. ( 1994; ). Molecular cloning, nucleotide sequence and expression of a Sulfolobus solfataricus gene encoding a class II fumarase. FEBS Lett 337, 93–98.[CrossRef]
    [Google Scholar]
  6. Cook, R. A. & Sanwal, B. D. ( 1969; ). Isocitrate dehydrogenase (NAD-specific) from Neurospora crassa. Methods Enzymol 13, 42–47.
    [Google Scholar]
  7. Dacks, J. B., Dyall, P. L., Embley, T. M. & van der Giezen, M. ( 2006; ). Hydrogenosomal succinyl-CoA synthetase from the rumen-dwelling fungus Neocallimastix patriciarum; an energy-producing enzyme of mitochondrial origin. Gene 373, 75–82.[CrossRef]
    [Google Scholar]
  8. Duggleby, R. G. & Dennis, D. T. ( 1974; ). Nicotinamide adenine dinucleotide-specific glyceraldehyde 3-phosphate dehydrogenase from Pisum sativum. J Biol Chem 249, 167–174.
    [Google Scholar]
  9. Dunn, L. A. & Boreham, P. F. ( 1991; ). The in vitro activity of drugs against Blastocystis hominis. J Antimicrob Chemother 27, 507–516.[CrossRef]
    [Google Scholar]
  10. Edwards, D. I. ( 1993; ). Nitroimidazole drugs-action and resistance mechanisms. II. Mechanisms of resistance. J Antimicrob Chemother 31, 201–210.[CrossRef]
    [Google Scholar]
  11. Embley, T. M., van der Giezen, M., Horner, D. S., Dyal, P. S. & Foster, P. ( 2003; ). Mitochondria and hydrogenosomes are two forms of the same fundamental organelle. Philos Trans R Soc Lond B Biol Sci 358, 191–201.[CrossRef]
    [Google Scholar]
  12. Englard, S. ( 1969; ). Extramitochondrial l-malate dehydrogenase of beef heart. Methods Enzymol 13, 123–129.
    [Google Scholar]
  13. Fansler, B. & Lowenstein, J. M. ( 1969; ). Aconitase from pig heart. Methods Enzymol 13, 26–30.
    [Google Scholar]
  14. Fukuda, W., Fukui, T., Atomi, H. & Imanaka, T. ( 2004; ). First characterization of an archaeal GTP-dependent phosphoenolpyruvate carboxykinase from the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. J Bacteriol 186, 4620–4627.[CrossRef]
    [Google Scholar]
  15. Genda, T., Watabe, S. & Ozaki, H. ( 2006; ). Purification and characterization from Corynebacterium glutamicum. Biosci Biotechnol Biochem 70, 1102–1109.[CrossRef]
    [Google Scholar]
  16. Gerbod, D., Edgcomb, V. P., Noël, C., Vanacova, S., Wintjens, R., Tachezy, J., Sogin, M. L. & Viscogliosi, E. ( 2001; ). Phylogenetic relationships of class II fumarase genes from trichomonads. Mol Biol Evol 18, 1574–1584.[CrossRef]
    [Google Scholar]
  17. Gorrell, T. E., Yarlett, N. & Müller, M. ( 1984; ). Isolation and characterization of Trichomonas vaginalis ferredoxin. Carlsberg Res Commun 49, 259–268.[CrossRef]
    [Google Scholar]
  18. Haresh, K., Suresh, K., Khairul Anus, A. & Saminathan, S. ( 1999; ). Isolate resistance of Blastocystis hominis to metronidazole. Trop Med Int Health 4, 274–277.[CrossRef]
    [Google Scholar]
  19. Hart, D. T. & Opperdoes, F. R. ( 1984; ). The occurrence of glycosomes (microbodies) in the promastigote stage of four major Leishmania species. Mol Biochem Parasitol 13, 159–172.[CrossRef]
    [Google Scholar]
  20. Hoffmeister, M., Piotrowski, M., Nowitzki, U. & Martin, W. ( 2005; ). Mitochondrial trans-2-enoyl-CoA reductase of wax ester fermentation from Euglena gracilis defines a new family of enzymes involved in lipid synthesis. J Biol Chem 280, 4329–4338.[CrossRef]
    [Google Scholar]
  21. Hrdy, I., Hirt, R. P., Dolezal, P., Bardonova, L., Foster, P. G., Tachezy, J. & Embley, T. M. ( 2004; ). Trichomonas hydrogenosomes contain the NADH dehydrogenase module of mitochondrial complex I. Nature 432, 618–622.[CrossRef]
    [Google Scholar]
  22. Hrdy, I., Cammack, R., Stopka, P., Kulda, J. & Tachezy, J. ( 2005; ). Alternative pathway of metronidazole activation in Trichomonas vaginalis hydrogenosomes. Antimicrob Agents Chemother 49, 5033–5036.[CrossRef]
    [Google Scholar]
  23. Inui, H., Ono, K., Miyatake, K., Nakano, Y. & Kitaoka, S. ( 1987; ). Purification and characterization of pyruvate : NADP+ oxidoreductase in Euglena gracilis. J Biol Chem 262, 9130–9135.
    [Google Scholar]
  24. Keithly, J. S., Langreth, S. G., Buttle, K. F. & Mannella, C. A. ( 2005; ). Electron tomographic and ultrastructural analysis of the Cryptosporidium parvum relict mitochondrion, its associated membranes, and organelles. J Eukaryot Microbiol 52, 132–140.[CrossRef]
    [Google Scholar]
  25. Krayl, M., Lim, J. H., Martin, F., Guiard, B. & Voos, W. ( 2007; ). A cooperative action of the ATP-dependent import motor complex and the inner membrane potential drives mitochondrial preprotein import. Mol Cell Biol 27, 411–425.[CrossRef]
    [Google Scholar]
  26. Lavier, G. ( 1952; ). Blastocystis spp. Ann Parasitol Hum Comp 27, 339–356.
    [Google Scholar]
  27. Lee, M. J. ( 1991; ). Pathogenicity of Blastocystis hominis. J Clin Microbiol 29, 2089
    [Google Scholar]
  28. Lindmark, D. G. & Müller, M. ( 1973; ). Hydrogenosome, a cytoplasmic organelle of the anaerobic flagellate Tritrichomonas foetus, and its role in pyruvate metabolism. J Biol Chem 248, 7724–7728.
    [Google Scholar]
  29. Lindmark, D. G. & Müller, M. ( 1974; ). Biochemical cytology of anaerobic flagellates. II. Subcellular distribution of oxidoreductases and hydrolases in Monocercomonas sp. J Protozool 21, 374–378.[CrossRef]
    [Google Scholar]
  30. Lindmark, D. G., Eckenrode, B. L., Halberg, L. A. & Dinbergs, I. D. ( 1989; ). Carbohydrate, energy and hydrogenosomal metabolism of Tritrichomonas foetus and Trichomonas vaginalis. J Protozool 36, 214–216.[CrossRef]
    [Google Scholar]
  31. Ma, Y.-C., Funk, M., Dunham, W. R. & Komuniecki, R. ( 1993; ). Purification and characterization of electron-transfer flavoprotein : rhodoquinone oxidoreductase from anaerobic mitochondria of the adult parasitic nematode Ascaris suum. J Biol Chem 268, 20360–20365.
    [Google Scholar]
  32. Marti, M., Regos, A., Li, Y., Schraner, E. M., Wild, P., Muller, N., Knopf, L. G. & Hehl, A. B. ( 2003; ). An ancestral secretory apparatus in the protozoan parasite Giardia intestinalis. J Biol Chem 278, 24837–24848.[CrossRef]
    [Google Scholar]
  33. Martin, W. ( 1999; ). A briefly argued case that mitochondria and plastids are descendents of endosymbionts, but that the nuclear compartment is not. Proc Biol Sci 266, 1387–1395.[CrossRef]
    [Google Scholar]
  34. Nanba, H., Takaoka, Y. & Hasegawa, J. ( 2003; ). Purification and characterization of formate dehydrogenase from Ancylobacter aquaticus strain KNK607M, and cloning of the gene. Biosci Biotechnol Biochem 67, 720–728.[CrossRef]
    [Google Scholar]
  35. Nasirudeen, A. M. & Tan, K. S. ( 2004; ). Isolation and characterization of the mitochondrion-like organelle from Blastocystis hominis. J Microbiol Methods 58, 101–109.[CrossRef]
    [Google Scholar]
  36. Noël, C., Dufernez, F., Gerbod, D., Edgcomb, V. P., Delgado-Viscogliosi, P., Ho, L. C., Singh, M., Wintjens, R., Sogin, M. L. & other authors ( 2005; ). Molecular phylogenies of Blastocystis isolates from different hosts: implications for genetic diversity, identification of species, and zoonosis. J Clin Microbiol 43, 348–355.[CrossRef]
    [Google Scholar]
  37. Parvin, R. ( 1969; ). Citrate synthase from yeast. Methods Enzymol 13, 16–19.
    [Google Scholar]
  38. Prasad, K. N., Nag, V. L., Dhole, T. N. & Ayyagari, A. ( 2000; ). Identification of enteric pathogens in HIV-positive patients with diarrhea in northern India. J Health Popul Nutr 18, 23–26.
    [Google Scholar]
  39. Prence, E. M. & Natowicz, M. R. ( 1992; ). Diagnosis of alpha mannosidosis by measuring alpha mannosidase in plasma. Clin Chem 38, 501–503.
    [Google Scholar]
  40. Puthia, M. K., Sio, S. W., Lu, J. & Tan, K. S. ( 2006; ). Blastocystis ratti induces contact-independent apoptosis, F-actin rearrangement, and barrier function disruption in IEC-6 cells. Infect Immun 74, 4114–4123.[CrossRef]
    [Google Scholar]
  41. Rotte, C., Stejskal, F., Zhu, G., Keithly, J. S. & Martin, W. ( 2001; ). Pyruvate : NADP+ oxidoreductase from the mitochondrion of Euglena gracilis and from the apicomplexan Cryptosporidium parvum: a biochemical relic linking pyruvate metabolism in mitochondriate and amitochondriate protists. Mol Biol Evol 18, 710–720.[CrossRef]
    [Google Scholar]
  42. Russo, A. R., Stone, S. L., Taplin, M. E., Snapper, H. J. & Doern, G. V. ( 1988; ). Presumptive evidence for Blastocystis hominis as a cause of colitis. Arch Intern Med 148, 1064 [CrossRef]
    [Google Scholar]
  43. Sanadi, D. R. ( 1969; ). α-Ketoglutarate dehydrogenase from pig heart. Methods Enzymol 13, 52–55.
    [Google Scholar]
  44. Singer, T. P. & Kearney, E. B. ( 1963; ). Succinate dehydrogenase. In The Enzymes, pp. 383–445. Edited by P. D. Boyer, K. H. Lardy & K. Myrback. New York: Academic Press.
  45. Steinbuchel, A. & Müller, M. ( 1986; ). Anaerobic pyruvate metabolism of Tritrichomonas foetus and Trichomonas vaginalis hydrogenosomes. Mol Biochem Parasitol 20, 57–65.[CrossRef]
    [Google Scholar]
  46. Stensvold, C. R., Suresh, G. K., Tan, K. S., Thompson, R. C., Traub, R. J., Viscogliosi, E., Yoshikawa, H. & Clark, C. G. ( 2007; ). Terminology for Blastocystis subtypes – a consensus. Trends Parasitol 23, 93–96.[CrossRef]
    [Google Scholar]
  47. Suzuki, T., Sato, M., Yoshida, T. & Tuboi, S. ( 1989; ). Rat liver mitochondrial and cytosolic fumarases with identical amino acid sequences are encoded from a single gene. J Biol Chem 264, 2581–2586.
    [Google Scholar]
  48. Tachezy, J., Sanchez, L. B. & Müller, M. ( 2001; ). Mitochondrial type iron-sulfur cluster assembly in the amitochondriate eukaryotes Trichomonas vaginalis and Giardia intestinalis, as indicated by the phylogeny of IscS. Mol Biol Evol 18, 1919–1928.[CrossRef]
    [Google Scholar]
  49. Tovar, J., Fischer, A. & Clark, C. G. ( 1999; ). The mitosome, a novel organelle related to mitochondria in the amitochondrial parasite Entamoeba histolytica. Mol Microbiol 32, 1013–1021.[CrossRef]
    [Google Scholar]
  50. van Grinsven, K. W., Rosnowsky, S., van Weelden, S. W., Pütz, S., van der Giezen, M., Martin, W., van Hellemond, J. J., Tielens, A. G. & Henze, K. ( 2008; ). Acetate : succinate CoA-transferase in the hydrogenosomes of Trichomonas vaginalis: identification and characterization. J Biol Chem 283, 1411–1418.[CrossRef]
    [Google Scholar]
  51. Van Hellemond, J. J., Opperdoes, F. R. & Tielens, A. G. ( 1998; ). Trypanosomatidae produce acetate via a mitochondrial acetate : succinate CoA transferase. Proc Natl Acad Sci U S A 95, 3036–3041.[CrossRef]
    [Google Scholar]
  52. Van Hellemond, J. J., Opperdoes, F. R. & Tielens, A. G. ( 2005; ). The extraordinary mitochondrion and unusual citric acid cycle in Trypanosoma brucei. Biochem Soc Trans 33, 967–971.[CrossRef]
    [Google Scholar]
  53. van Weelden, S. W. H., van Hellemond, J. J., Opperdoes, F. R. & Tielens, A. G. M. ( 2005; ). New functions for parts of the krebs cycle in procyclic Trypanosoma brucei, a cycle not operating as a cycle. J Biol Chem 280, 12451–12460.[CrossRef]
    [Google Scholar]
  54. Vannatta, J. B., Adamson, D. & Mullican, K. ( 1985; ). Blastocystis hominis infection presenting as recurrent diarrhea. Ann Intern Med 102, 495–496.[CrossRef]
    [Google Scholar]
  55. Walderich, B., Bernauer, S., Renner, M., Knobloch, J. & Burchard, G. D. ( 1998; ). Cytopathic effects of Blastocystis hominis on Chinese hamster ovary (CHO) and adenocarcinoma HT29 cell cultures. Trop Med Int Health 3, 385–390.[CrossRef]
    [Google Scholar]
  56. Weg, A. L., Soave, R. & Jacobson, I. M. ( 1987; ). The significance of intestinal Blastocystis hominis infection. Gastroenterology 92, 1688
    [Google Scholar]
  57. Woods, S. A., Schwartzbach, S. D. & Guest, J. D. ( 1988; ). Two biochemically distinct classes of fumarase in Escherichia coli. Biochim Biophys Acta 954, 14–26.[CrossRef]
    [Google Scholar]
  58. Yarlett, N., Hann, A. C., Lloyd, D. & Williams, A. G. ( 1981; ). Hydrogenosomes in the rumen protozoan Dasytricha ruminantium Schuberg. Biochem J 200, 365–372.
    [Google Scholar]
  59. Yarlett, N., Orpin, C. G., Munn, E. A., Yarlett, N. C. & Greenwood, C. A. ( 1986; ). Hydrogenosomes in the rumen fungus Neocallimastix patriciarum. Biochem J 236, 729–739.
    [Google Scholar]
  60. Zierdt, C. H. ( 1986; ). Cytochrome free mitochondria of an anaerobic protozoan – Blastocystis hominis. J Protozool 33, 67–69.[CrossRef]
    [Google Scholar]
  61. Zierdt, C. H., Swan, J. C. & Hosseini, J. ( 1983; ). In vitro response of Blastocystis hominis to antiprotozoal drugs. J Protozool 30, 332–334.[CrossRef]
    [Google Scholar]
  62. Zierdt, C. H., Donnolley, C. T., Muller, J. & Constantopoulos, G. ( 1988; ). Biochemical and ultrastructural study of Blastocystis hominis. J Clin Microbiol 26, 965–970.
    [Google Scholar]
  63. Zuckerman, M. J., Watts, M. T., Ho, H. & Meriano, F. V. ( 1994; ). Blastocystis hominis infection and intestinal injury. Am J Med Sci 308, 96–101.[CrossRef]
    [Google Scholar]
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