Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Branched tricarboxylic acid metabolism in Plasmodium falciparum

Nature volume 466, pages 774–778 (2010)Cite this article

Subjects

A Retraction to this article was published on 24 April 2013

A Corrigendum to this article was published on 19 January 2011

Abstract

A central hub of carbon metabolism is the tricarboxylic acid cycle1, which serves to connect the processes of glycolysis, gluconeogenesis, respiration, amino acid synthesis and other biosynthetic pathways. The protozoan intracellular malaria parasites (Plasmodium spp.), however, have long been suspected of possessing a significantly streamlined carbon metabolic network in which tricarboxylic acid metabolism plays a minor role2. Blood-stage Plasmodium parasites rely almost entirely on glucose fermentation for energy and consume minimal amounts of oxygen3, yet the parasite genome encodes all of the enzymes necessary for a complete tricarboxylic acid cycle4. Here, by tracing 13C-labelled compounds using mass spectrometry5 we show that tricarboxylic acid metabolism in the human malaria parasite Plasmodium falciparum is largely disconnected from glycolysis and is organized along a fundamentally different architecture from the canonical textbook pathway. We find that this pathway is not cyclic, but rather is a branched structure in which the major carbon sources are the amino acids glutamate and glutamine. As a consequence of this branched architecture, several reactions must run in the reverse of the standard direction, thereby generating two-carbon units in the form of acetyl-coenzyme A. We further show that glutamine-derived acetyl-coenzyme A is used for histone acetylation, whereas glucose-derived acetyl-coenzyme A is used to acetylate amino sugars. Thus, the parasite has evolved two independent production mechanisms for acetyl-coenzyme A with different biological functions. These results significantly clarify our understanding of the Plasmodium metabolic network and highlight the ability of altered variants of central carbon metabolism to arise in response to unique environments.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Glutamine drives reverse flux through the TCA cycle.
Figure 2: Acetyl groups deriving from glucose and glutamine are functionally distinct.
Figure 3: Malate excretion by P. falciparum -infected red blood cell cultures.
Figure 4: An integrated model for central carbon metabolism in P. falciparum.

Similar content being viewed by others

References

  1. Krebs, H. A. & Johnson, W. A. The role of citric acid in intermediate metabolism in animal tissues. Enzymologia 4, 148–156 (1937)

    CAS  Google Scholar 

  2. van Dooren, G. G., Stimmler, L. M. & McFadden, G. I. Metabolic maps and functions of the Plasmodium mitochondrion. FEMS Microbiol. Rev. 30, 596–630 (2006)

    Article  CAS  PubMed  Google Scholar 

  3. Sherman, I. W. in Malaria, Parasite Biology, Pathogenesis and Protection (ed. Sherman, I. W.) 135–143 (ASM, 1998)

    Google Scholar 

  4. Gardner, M. J. et al. Genome sequence of the human malaria parasite Plasmodium falciparum . Nature 419, 498–511 (2002)

    Article  ADS  CAS  PubMed  Google Scholar 

  5. Munger, J. et al. Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy. Nature Biotechnol. 26, 1179–1186 (2008)

    Article  CAS  Google Scholar 

  6. Vaidya, A. B. & Mather, M. W. Mitochondrial evolution and functions in malaria parasites. Annu. Rev. Microbiol. 63, 249–267 (2009)

    Article  CAS  PubMed  Google Scholar 

  7. Painter, H. J., Morrisey, J. M., Mather, M. W. & Vaidya, A. B. Specific role of mitochondrial electron transport in blood-stage Plasmodium falciparum . Nature 446, 88–91 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  8. Bozdech, Z. et al. The transcriptome of the intraerythrocytic developmental cycle of Plasmodium falciparum . PLoS Biol. 1, e5 (2003)

    Article  PubMed  PubMed Central  Google Scholar 

  9. Tonkin, C. J. et al. Localization of organellar proteins in Plasmodium falciparum using a novel set of transfection vectors and a new immunofluorescence fixation method. Mol. Biochem. Parasitol. 137, 13–21 (2004)

    Article  CAS  PubMed  Google Scholar 

  10. Hodges, M. et al. An iron regulatory-like protein expressed in Plasmodium falciparum displays aconitase activity. Mol. Biochem. Parasitol. 143, 29–38 (2005)

    Article  CAS  PubMed  Google Scholar 

  11. Wrenger, C. & Muller, S. Isocitrate dehydrogenase of Plasmodium falciparum . Eur. J. Biochem. 270, 1775–1783 (2003)

    Article  CAS  PubMed  Google Scholar 

  12. Suraveratum, N. et al. Purification and characterization of Plasmodium falciparum succinate dehydrogenase. Mol. Biochem. Parasitol. 105, 215–222 (2000)

    Article  CAS  PubMed  Google Scholar 

  13. Olszewski, K. L. et al. Host-parasite interactions revealed by Plasmodium falciparum metabolomics. Cell Host Microbe 5, 191–199 (2009)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Foth, B. J. et al. The malaria parasite Plasmodium falciparum has only one pyruvate dehydrogenase complex, which is located in the apicoplast. Mol. Microbiol. 55, 39–53 (2005)

    Article  CAS  PubMed  Google Scholar 

  15. Blum, J. J. & Ginsburg, H. Absence of α-ketoglutarate dehydrogenase activity and presence of CO2-fixing activity in Plasmodium falciparum grown in vitro in human erythrocytes. J. Protozool. 31, 167–169 (1984)

    Article  CAS  PubMed  Google Scholar 

  16. Elford, B. C., Haynes, J. D., Chulay, J. D. & Wilson, R. J. Selective stage-specific changes in the permeability to small hydrophilic solutes of human erythrocytes infected with Plasmodium falciparum . Mol. Biochem. Parasitol. 16, 43–60 (1985)

    Article  CAS  PubMed  Google Scholar 

  17. Liu, J. et al. Plasmodium falciparum ensures its amino acid supply with multiple acquisition pathways and redundant proteolytic enzyme systems. Proc. Natl Acad. Sci. USA 103, 8840–8845 (2006)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  18. Yoo, H., Antoniewicz, M. R., Stephanopoulos, G. & Kelleher, J. K. Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line. J. Biol. Chem. 283, 20621–20627 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Vaughan, A. M. et al. Type II fatty acid synthesis is essential only for malaria parasite late liver stage development. Cell. Microbiol. 11, 506–520 (2009)

    Article  CAS  PubMed  Google Scholar 

  20. Yu, M. et al. The fatty acid biosynthesis enzyme FabI plays a key role in the development of liver-stage malarial parasites. Cell Host Microbe 4, 567–578 (2008)

    Article  PubMed  PubMed Central  Google Scholar 

  21. Gowda, D. C. & Davidson, E. A. Protein glycosylation in the malaria parasite. Parasitol. Today 15, 147–152 (1999)

    Article  CAS  PubMed  Google Scholar 

  22. Downie, M. J., Kirk, K. & Mamoun, C. B. Purine salvage pathways in the intraerythrocytic malaria parasite Plasmodium falciparum . Eukaryot. Cell 7, 1231–1237 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Fry, M. & Beesley, J. E. Mitochondria of mammalian Plasmodium spp . Parasitology 102, 17–26 (1991)

    Article  PubMed  Google Scholar 

  24. Wellen, K. E. et al. ATP-citrate lyase links cellular metabolism to histone acetylation. Science 324, 1076–1080 (2009)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  25. Wang, Q. et al. Acetylation of metabolic enzymes coordinates carbon source utilization and metabolic flux. Science 327, 1004–1007 (2010)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  26. Zhao, S. et al. Regulation of cellular metabolism by protein lysine acetylation. Science 327, 1000–1004 (2010)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  27. Daily, J. P. et al. Distinct physiological states of Plasmodium falciparum in malaria-infected patients. Nature 450, 1091–1095 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  28. Lasonder, E. et al. Proteomic profiling of Plasmodium sporozoite maturation identifies new proteins essential for parasite development and infectivity. PLoS Pathog. 4, e1000195 (2008)

    Article  PubMed  PubMed Central  Google Scholar 

  29. Mather, M. W., Henry, K. W. & Vaidya, A. B. Mitochondrial drug targets in apicomplexan parasites. Curr. Drug Targets 8, 49–60 (2007)

    Article  CAS  PubMed  Google Scholar 

  30. Andrews, K. T., Tran, T. N., Wheatley, N. C. & Fairlie, D. P. Targeting histone deacetylase inhibitors for anti-malarial therapy. Curr. Top. Med. Chem. 9, 292–308 (2009)

    Article  CAS  PubMed  Google Scholar 

  31. Trager, W. & Jensen, J. B. Human malaria parasites in continuous culture. Science 193, 673–675 (1976)

    Article  ADS  CAS  PubMed  Google Scholar 

  32. Lambros, C. & Vanderberg, J. P. Synchronization of Plasmodium falciparum erythrocytic stages in culture. J. Parasitol. 65, 418–420 (1979)

    Article  CAS  PubMed  Google Scholar 

  33. Moore, G. E., Gerner, R. E. & Franklin, H. A. Culture of normal human leukocytes. J. Am. Med. Assoc. 199, 519–524 (1967)

    Article  CAS  Google Scholar 

  34. Lu, W., Bennett, B. D. & Rabinowitz, J. D. Analytical strategies for LC–MS-based targeted metabolomics. J. Chromatogr. B 871, 236–242 (2008)

    Article  CAS  Google Scholar 

  35. Keller, A. et al. A uniform proteomics MS/MS analysis platform utilizing open XML file formats. Mol. Syst. Biol. 1, 2005.0017 (2005)

    Article  PubMed  PubMed Central  Google Scholar 

  36. Breiman, L. Random forests. Mach. Learn. 45, 5–32 (2001)

    Article  MATH  Google Scholar 

  37. Mi-Ichi, F., Kita, K. & Mitamura, T. Intraerythrocytic Plasmodium falciparum utilize a broad range of serum-derived fatty acids with limited modification for their growth. Parasitology 133, 399–410 (2006)

    Article  CAS  PubMed  Google Scholar 

  38. Miao, J., Fan, Q., Cui, L. & Li, J. The malaria parasite Plasmodium falciparum histones: organization, expression, and acetylation. Gene 369, 53–65 (2006)

    Article  CAS  PubMed  Google Scholar 

  39. Shechter, D., Dormann, H. L., Allis, C. D. & Hake, S. B. Extraction, purification and analysis of histones. Nature Protocols 2, 1445–1457 (2007)

    Article  CAS  PubMed  Google Scholar 

  40. Garcia, B. A. et al. Chemical derivatization of histones for facilitated analysis by mass spectrometry. Nature Protocols 2, 933–938 (2007)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Takashima, E. et al. Isolation of mitochondria from Plasmodium falciparum showing dihydroorotate dependent respiration. Parasitol. Int. 50, 273–278 (2001)

    Article  CAS  PubMed  Google Scholar 

  42. Kornberg, A. & Pricer, W. E., Jr Di- and triphosphopyridine nucleotide isocitric dehydrogenases in yeast. J. Biol. Chem. 189, 123–136 (1951)

    CAS  PubMed  Google Scholar 

  43. Ma, Z., Chu, C. H. & Cheng, D. A novel direct homogeneous assay for ATP citrate lyase. J. Lipid Res. 50, 2131–2135 (2009)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Bergmeyer, H. U., Gawehn, K. & Grassl, M. in Methods of Enzymatic Analysis Vol. 1 (ed. Bergmeyer, H. U.) 442–443 (Academic, 1974)

    Google Scholar 

  45. Kadekoppala, M., Kline, K., Akompong, T. & Haldar, K. Stable expression of a new chimeric fluorescent reporter in the human malaria parasite Plasmodium falciparum . Infect. Immun. 68, 2328–2332 (2000)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. O'Donnell, R. A. et al. A genetic screen for improved plasmid segregation reveals a role for Rep20 in the interaction of Plasmodium falciparum chromosomes. EMBO J. 21, 1231–1239 (2002)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Fidock, D. A. & Wellems, T. E. Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil. Proc. Natl Acad. Sci. USA 94, 10931–10936 (1997)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  48. Forer, A. & Pickett-Heaps, J. D. Cytochalasin D and latrunculin affect chromosome behaviour during meiosis in crane-fly spermatocytes. Chromosome Res. 6, 533–549 (1998)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank G. McFadden and I. Sherman for discussions and scrutiny of the manuscript; B. Bennett, T. Campbell, E. De Silva, J. O’Hara, and H. Painter for reading of the manuscript; I. Ying for assistance with histone extraction; T. Spurck and C. Tonkin for the modified erythrocyte immobilization procedure for microscopy; M. Clasquin and W. Lu for developing the LC–MS methodology; E. Melamud for LC–MS data extraction and analysis; and J. Groves and H. Cooper for GC–MS analysis. M.L. is funded by the Burroughs Wellcome Fund and an NIH Director’s New Innovators award (1DP2OD001315-01). J.D.R. is funded by a Beckman Young Investigators award, an NSF CAREER award and NIH R01 AI078063. M.L and J.D.R. receive support from the Center for Quantitative Biology (P50 GM071508). B.A.G. receives support from NSF grant CBET-0941143. K.L.O. is funded by an NSF Graduate Research Fellowship. J.M.M., M.W.M. and A.B.V. are supported by grant AI028398 from NIAID, NIH.

Author information

Authors and Affiliations

  1. Department of Molecular Biology and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, 08544, New Jersey, USA

    Kellen L. Olszewski & Manuel Llinás

  2. Center for Molecular Parasitology, Drexel University College of Medicine, Philadelphia, 19129, Pennsylvania, USA

    Michael W. Mather, Joanne M. Morrisey & Akhil B. Vaidya

  3. Department of Molecular Biology, Princeton University, Princeton, 08544, New Jersey, USA

    Benjamin A. Garcia

  4. Department of Chemistry and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, 08544, New Jersey, USA

    Joshua D. Rabinowitz

Authors
  1. Kellen L. Olszewski
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael W. Mather
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joanne M. Morrisey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Benjamin A. Garcia
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Akhil B. Vaidya
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Joshua D. Rabinowitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Manuel Llinás
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

K.L.O. cultured the parasites, and collected and analysed all LC–MS and GC–MS data; B.A.G. performed mass spectrometric analysis of histones. M.W.M. and J.M.M. carried out IDH localization studies. M.W.M. purified mitochondria and K.L.O. did biochemical assays. K.L.O., M.L., J.D.R., M.W.M., A.B.V. and B.A.G. designed the study; J.D.R. provided the metabolomic technology. M.L. and K.L.O. wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Manuel Llinás.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-8 with legends, a Supplementary Discussion and References. Supplementary Fig. 3 was corrected on 20 January 2011. (PDF 4015 kb)

PowerPoint slides

Rights and permissions

Reprints and permissions

About this article

Cite this article

Olszewski, K., Mather, M., Morrisey, J. et al. Branched tricarboxylic acid metabolism in Plasmodium falciparum. Nature 466, 774–778 (2010). https://doi.org/10.1038/nature09301

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature09301

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing