Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose.
Metabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of Trypanosoma brucei, a parasite responsible for human African trypanosomiasis. Glucose enters many branches of metabolism beyond glycolysi...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-03-01
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| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1004689 |
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| author | Darren J Creek Muriel Mazet Fiona Achcar Jana Anderson Dong-Hyun Kim Ruwida Kamour Pauline Morand Yoann Millerioux Marc Biran Eduard J Kerkhoven Achuthanunni Chokkathukalam Stefan K Weidt Karl E V Burgess Rainer Breitling David G Watson Frédéric Bringaud Michael P Barrett |
| author_facet | Darren J Creek Muriel Mazet Fiona Achcar Jana Anderson Dong-Hyun Kim Ruwida Kamour Pauline Morand Yoann Millerioux Marc Biran Eduard J Kerkhoven Achuthanunni Chokkathukalam Stefan K Weidt Karl E V Burgess Rainer Breitling David G Watson Frédéric Bringaud Michael P Barrett |
| author_sort | Darren J Creek |
| collection | DOAJ |
| description | Metabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of Trypanosoma brucei, a parasite responsible for human African trypanosomiasis. Glucose enters many branches of metabolism beyond glycolysis, which has been widely held to be the sole route of glucose metabolism. Whilst pyruvate is the major end-product of glucose catabolism, its transamination product, alanine, is also produced in significant quantities. The oxidative branch of the pentose phosphate pathway is operative, although the non-oxidative branch is not. Ribose 5-phosphate generated through this pathway distributes widely into nucleotide synthesis and other branches of metabolism. Acetate, derived from glucose, is found associated with a range of acetylated amino acids and, to a lesser extent, fatty acids; while labelled glycerol is found in many glycerophospholipids. Glucose also enters inositol and several sugar nucleotides that serve as precursors to macromolecule biosynthesis. Although a Krebs cycle is not operative, malate, fumarate and succinate, primarily labelled in three carbons, were present, indicating an origin from phosphoenolpyruvate via oxaloacetate. Interestingly, the enzyme responsible for conversion of phosphoenolpyruvate to oxaloacetate, phosphoenolpyruvate carboxykinase, was shown to be essential to the bloodstream form trypanosomes, as demonstrated by the lethal phenotype induced by RNAi-mediated downregulation of its expression. In addition, glucose derivatives enter pyrimidine biosynthesis via oxaloacetate as a precursor to aspartate and orotate. |
| format | Article |
| id | doaj-art-6f5a415b91f34ce6b29b7e4fca138d7b |
| institution | OA Journals |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2015-03-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-6f5a415b91f34ce6b29b7e4fca138d7b2025-08-20T02:22:38ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742015-03-01113e100468910.1371/journal.ppat.1004689Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose.Darren J CreekMuriel MazetFiona AchcarJana AndersonDong-Hyun KimRuwida KamourPauline MorandYoann MilleriouxMarc BiranEduard J KerkhovenAchuthanunni ChokkathukalamStefan K WeidtKarl E V BurgessRainer BreitlingDavid G WatsonFrédéric BringaudMichael P BarrettMetabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of Trypanosoma brucei, a parasite responsible for human African trypanosomiasis. Glucose enters many branches of metabolism beyond glycolysis, which has been widely held to be the sole route of glucose metabolism. Whilst pyruvate is the major end-product of glucose catabolism, its transamination product, alanine, is also produced in significant quantities. The oxidative branch of the pentose phosphate pathway is operative, although the non-oxidative branch is not. Ribose 5-phosphate generated through this pathway distributes widely into nucleotide synthesis and other branches of metabolism. Acetate, derived from glucose, is found associated with a range of acetylated amino acids and, to a lesser extent, fatty acids; while labelled glycerol is found in many glycerophospholipids. Glucose also enters inositol and several sugar nucleotides that serve as precursors to macromolecule biosynthesis. Although a Krebs cycle is not operative, malate, fumarate and succinate, primarily labelled in three carbons, were present, indicating an origin from phosphoenolpyruvate via oxaloacetate. Interestingly, the enzyme responsible for conversion of phosphoenolpyruvate to oxaloacetate, phosphoenolpyruvate carboxykinase, was shown to be essential to the bloodstream form trypanosomes, as demonstrated by the lethal phenotype induced by RNAi-mediated downregulation of its expression. In addition, glucose derivatives enter pyrimidine biosynthesis via oxaloacetate as a precursor to aspartate and orotate.https://doi.org/10.1371/journal.ppat.1004689 |
| spellingShingle | Darren J Creek Muriel Mazet Fiona Achcar Jana Anderson Dong-Hyun Kim Ruwida Kamour Pauline Morand Yoann Millerioux Marc Biran Eduard J Kerkhoven Achuthanunni Chokkathukalam Stefan K Weidt Karl E V Burgess Rainer Breitling David G Watson Frédéric Bringaud Michael P Barrett Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. PLoS Pathogens |
| title | Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. |
| title_full | Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. |
| title_fullStr | Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. |
| title_full_unstemmed | Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. |
| title_short | Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose. |
| title_sort | probing the metabolic network in bloodstream form trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose |
| url | https://doi.org/10.1371/journal.ppat.1004689 |
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