Journal of Molecular Evolution

, Volume 41, Issue 3, pp 388–396 | Cite as

Primary structure and eubacterial relationships of the pyruvate:Ferredoxin oxidoreductase of the amitochondriate eukaryoteTrichomonas vaginalis

  • Ivan Hrdý
  • Miklós Müller


In the eukaryotic unicellular organismTrichomonas vaginalis a key step of energy metabolism, the oxidative decarboxylation of pyruvate with the formation of acetyl-CoA, is catalyzed by the iron-sulfur protein pyruvate:ferredoxin oxidoreductase (PFO) and not by the almost-ubiquitous pyruvate dehydrogenase multienzyme complex. This enzyme is localized in the hydrogenosome, an organelle bounded by a double membrane. PFO and its closely related homolog, pyruvate: flavodoxin oxidoreductase, are enzymes found in a number of archaebacteria and eubacteria. The presence of these enzymes in eukaryotes is restricted, however, to a few amitochondriate groups. To gain more insight into the evolutionary relationships ofT. vaginalis PFO we determined the primary structure of its two genes (pfoA andpfoB). The deduced amino acid sequences showed 95% positional identity. Motifs implicated in related enzymes in liganding the Fe-S centers and thiamine pyrophosphate were well conserved. TheT. vaginalis PFOs were found to be homologous to eubacterial pyruvate: flavodoxin oxidoreductases and showed about 40% amino acid identity to these enzymes over their entire length. Lack of eubacterial PFO sequences precluded a comparison.pfoA andpfoB revealed a greater distance from related enzymes of Archaebacteria. The conceptual translation of the nucleotide sequences predicted an amino-terminal pentapeptide not present in the mature protein. This processed leader sequence was similar to but shorter than leader sequences noted in other hydrogenosomal proteins. These sequences are assumed to be involved in organellar targeting and import. The results underscore the unusual characteristics ofT. vaginalis metabolism and of their hydrogenosomes. They also suggest that in its energy metabolismT. vaginalis is closer to eubacteria than archaebacteria.

Key words

Hydrogenosome Molecular phylogeny Anaerobic protist 



DNA polymerase chain reaction


pyruvate dehydrogenase


pyruvate:ferredoxin oxidoreductase


thiamine pyrophosphate


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410Google Scholar
  2. Arnold W, Rump A, Klipp W, Priefer U, Puhler A (1988) Nucleotide sequence of a 24,206-base-pair DNA fragment carrying the entire nitrogen fixiation gene cluster ofKlebsiella pneumoniae. J Mol Biol 203:715–738Google Scholar
  3. Bauer CC, Scappino L, Haselkorn R (1993) Growth of the cyanobacteriumAnabaena on molecular nitrogen:NifJ is required when iron is limited. Proc Natl Acad Sci USA 90:8812–8816Google Scholar
  4. Blarney JM, Adams MWW (1993) Purification and characterization of pyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeonPyrococcus furiosus. Biochim Biophys Acta 1161:19–27Google Scholar
  5. Cabot EL, Beckenbach AT (1989) Simultaneous editing of multiple nucleic acid and protein sequences with ESEE. Comput Appl Biosci 5:233–234Google Scholar
  6. Cammack R, Kerscher L, Oesterhelt D (1980) A stable free radical intermediate in the reaction of 2-oxoacid:ferredoxin oxidoreductases ofHalobacterium halobium. FEBS Lett 118:271–273Google Scholar
  7. Cannon M, Cannon F, Buchanan-Wollaston V, Alley D, Alley A, Beynon J (1988) The nucleotide sequence of thenifJ gene ofKlebsiella pneumoniae. Nucleic Acids Res 16:11379Google Scholar
  8. Chapman A, Cammack R, Linstead DJ, Lloyd D (1986) Respiration ofTrichomonas vaginalis. Components detected by electron paramagnetic resonance spectroscopy. Eur J Biochem 156:193–198Google Scholar
  9. Coombs GH North MJ (1983) An analysis of the proteinases ofTrichomonas vaginalis by polyacrylamide gel electrophoresis. Parasitology 86:1–6Google Scholar
  10. Docampo R, Moreno SNJ, Mason RP (1987) Free radical intermediates in the reaction of pyruvateferredoxin oxidoreductase inTritrichomonas foetus hydrogenosomes. J Biol Chem 262:12417–12420Google Scholar
  11. Dore J, Stahl DA (1991) Phylogeny of anaerobic rumen Chytridiomycetes inferred from small subunit ribosomal RNA sequence comparisons. Can J Bot 69:1964–1971Google Scholar
  12. Embley TM, Finlay BJ (1994) The use of small subunit rRNA sequences to unravel the relationships between anaerobic ciliates and their methanogen endosymbionts. Microbiology 140:225–235Google Scholar
  13. Fenchel T, Finlay BJ (1991) The biology of free-living anaerobic ciliates. Eur J Protistol 26:201–215Google Scholar
  14. Gorrell TE, Yarlett N, Müller M (1984) Isolation and characterization ofTrichomonas vaginalis ferredoxin. Carlsberg Res Commun 49:259–268Google Scholar
  15. Hawkins CF, Borges A, Perham RN (1989) A common structural motif in thiamin pyrophosphate-binding enzymes. FEBS Lett 255:77–82Google Scholar
  16. Higgins DG (1994) CLUSTAL V: multiple alignment of DNA of protein sequences. Methods Mol Biol 25:307–318Google Scholar
  17. Hrdý I, Mertens E (1993) Purification and partial characterization of malate dehydrogenase (decarboxylating) fromTritrichomonas foetus hydrogenosomes. Parasitology 107:379–385Google Scholar
  18. Hrdý I, Müller M (1995) Primary structure of the hydrogenosomal malic enzyme ofTrichomonas vaginalis and its relationship to homologous enzymes. J Eukar Microbiol (in press)Google Scholar
  19. Inui H, Miyatake K, Nakano Y, Kitaoka S (1984) Occurrence of oxygen-sensitive, NADP+-dependent pyruvate dehydrogenase in mitochondria ofEuglena gracilis. J Biochem 96:931–934Google Scholar
  20. Inui H, Ono K, Miyatake K, Nakano Y, Kitaoka S (1987) Purification and characterization of pyruvate:NADP+ oxidoreductase inEuglena gracilis. J Biol Chem 262:9130–9135Google Scholar
  21. Johnson PJ, d'Oliveira CE, Gorrell TE, Müller M (1990) Molecular analysis of the hydrogenosomal ferredoxin of the anaerobic protist,Trichomonas vaginalis. Proc Natl Acad Sci USA 87:6097–6101Google Scholar
  22. Johnson PJ, Lahti CJ, Bradley PJ (1993) Biogenesis of the hydrogenosome in the anaerobic protistTrichomonas vaginalis. J Parasitol 79:664–670Google Scholar
  23. Kerscher L, Oesterhelt D (1981) The catalytic mechanism of 2-oxoacid:ferredoxin oxidoreductases fromHalobacterium halobium. One-electron transfer at two distinct steps of the catalytic cycle. Eur J Biochem 116:595–600Google Scholar
  24. Kerscher L, Oesterhelt D (1982) Pyruvate:ferredoxin oxidoreductase new findings on an ancient enzyme. Trends Biochem Sci 7:371–374Google Scholar
  25. Kreutzer R, Dayananda S, Klingmuller W (1991) Cotranscription of the electron transport protein genesnifJ andnifF inEnterobacter agglomerans 333. J Bacteriol 173:3252–3256Google Scholar
  26. Lahti CJ, d'Oliveira CE, Johnson PJ (1992) β-Succinyl-coenzyme A synthetase fromTrichomonas vaginalis is a soluble hydrogenosomal protein with an amino-terminal sequence that resembles mitochondrial presequences. J Bacteriol 174:6822–6830Google Scholar
  27. Lahti CJ, Bradley PJ, Johnson PJ (1994) Molecular characterization of the α-subunit ofTrichomonas vaginalis hydrogenosomal succinyl CoA synthetase. Mol Biochem Parasitol 66:309–318Google Scholar
  28. Länge S, Rozario C, Müller M (1994) Primary structure of the hydrogenosomal adenylate kinase ofTrichomonas vaginalis and its phylogenetic relationships. Mol Biochem Parasitol 66:297–308Google Scholar
  29. Leipe DD, Gunderson JH, Nerad TA, Sogin ML (1993) Small subunit ribosomal RNA+ ofHexamita infata and the quest for the first branch in the eukaryotic tree. Mol Biochem Parasitol 59:41–48Google Scholar
  30. Lindmark DG, Müller M (1973) Hydrogenosome, a cytoplasmic organelle of the anaerobic flagellate,Tritrichomonas foetus, and its role in pyruvate metabolism. J Biol Chem 248:7724–7728Google Scholar
  31. Lindmark DG, Müller M, Shio H (1975) Hydrogenosomes inTrichomonas vaginalis. J Parasitol 61:552–554Google Scholar
  32. Lindmark DG (1980) Energy metabolism of the anaerobic protozoonGiardia lamblia. Mol Biochem Parasitol 1:1–12Google Scholar
  33. Mai X, Adams MWW (1994) Indolepyruvate ferredoxin oxidoreductase from the hyperthermophilic archaeonPyrococcus furiosus. A new enzyme involved in peptide fermentation. J Biol Chem 269:16726–16732Google Scholar
  34. Markoš A, Miretsky A, Müller M (1993) A glyceraldehyde 3-phosphate dehydrogenase with eubacterial features in the amitochondriate eukaryote,Trichomonas vaginalis. J Mol Evol 37:631–643Google Scholar
  35. Mendis AHW, Schofield PJ (1994) Discussants report: giardia biochemistry. In: Thompson RCA, Reynoldson JA, Lymbery AJ (eds) Giardia: from molecules to disease. CAB International, Wallingford, UK, pp 205–213Google Scholar
  36. Müller M (1988) Energy metabolism of protozoa without mitochondria. Annu Rev Microbiol 42:465–488Google Scholar
  37. Müller M (1993) The hydrogenosome. J Gen Microbiol 139:2879–2889Google Scholar
  38. Payne MJ, Chapman A, Cammack R (1993) Evidence for an [Fe]-type hydrogenase in the parasitic protozoanTrichomonas vaginalis. FEBS Lett 317:101–104Google Scholar
  39. Philippe H (1993) MUST, a computer package of management utilities for sequences and trees. Nucleic Acids Res 21:5264–5272Google Scholar
  40. Plaga W, Lottspeich F, Oesterhelt D (1992) Improved purification, crystallization and primary structure of pyruvate:ferredoxin oxidoreductase fromHalobacterium halobium. Eur J Biochem 205:391–397Google Scholar
  41. Quon DVK, Delgadillo MG, Khachi A, Smale ST, Johnson PJ (1994) Similarity between a ubiquitous promoter element in an ancient eukaryote and mammalian initiator elements. Proc Natl Acad Sci USA 91:4579–4583Google Scholar
  42. Reed LJ (1974) Multienzyme complexes. Acc Chem Res 7:40–46Google Scholar
  43. Reeves RE, Warren LG, Susskind B, Lo H-S (1977) An energy conserving pyruvate-to-acetate pathway inEntamoeba histolytica. Pyruvate synthase and a new acetate thiokinase. J Biol Chem 252:726–731Google Scholar
  44. Reeves RE (1984) Metabolism ofEntamoeba histolytica Schaudinn, 1903. Adv Parasitol 23:105–142Google Scholar
  45. Smith ET, Blarney JM, Adams MWW (1994) Pyruvate ferredoxin oxidoreductases of the hyperthermophilic archaeon,Pyrococcus furiosus, and thehyperthermophilic bacterium,Thermotoga maritima, have different catalytic mechanisms. Biochemistry 33:1008–1016Google Scholar
  46. Sogin ML (1991) Early evolution and the origin of eukaryotes. Curr Opin Gen Dev 1:457–463Google Scholar
  47. Thauer RK, Jungermann K, Decker K (1977) Energy conservation in chemotrophic anaerobic bacteria. Bacteriol Rev 41:100–180Google Scholar
  48. Van Hejine G, Steppuhn J, Herrmann RG (1989) Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem 180:535–545Google Scholar
  49. Viscogliosi E, Philippe H, Baroin A, Perasso R, Brugerolle G (1993) Phylogeny of trichomonads based on partial sequences of large subunit rRNA and on cladistic analysis of morphological data. J Eukar Microbiol 40:411–421Google Scholar
  50. Wahl RC, Orme-Johnson WH (1987) Clostridial pyruvate oxidoreductase and the pyruvate-oxidizing enzyme specific for nitrogen fixation inKlebsiella pneumoniae are similar enzymes. J Biol Chem 262:10489–10496Google Scholar
  51. Whatley JM, John P, Whatley FR (1979) From extracellular to intracellular: the establishment of mitochondria and chloroplasts. Proc R Soc Lond [Biol] 204:165–187Google Scholar
  52. Wieland OH (1983) The mammalian pyruvate dehydrogenase complex: structure and regulation. Rev Biochem Physiol Pharmacol 96:123–170Google Scholar
  53. Williams K, Lowe PN, Leadlay PF (1987) Purification and characterization of pyruvate:ferredoxin oxidoreductase from the anaerobic protozoonTrichomonas vaginalis. Biochem J 246:529–536Google Scholar
  54. Yarlett N, Hann AC, Lloyd D, Williams A (1981) Hydrogenosomes in the rumen protozoonDasytricha ruminantium Schuberg. Biochem J 200:365–372Google Scholar
  55. Yarlett N, Orpin CG, Munn EA, Yarlett NC, Greenwood CA (1986) Hydrogenosomes in the rumen fungusNeocallimastix patriciarum. Biochem J 236:729–739Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Ivan Hrdý
    • 1
  • Miklós Müller
    • 1
  1. 1.The Rockefeller UniversityNew YorkUSA

Personalised recommendations