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Probing the Metabolic Network in Bloodstream-Form Trypanosoma brucei Using Untargeted Metabolomics with Stable Isotope Labelled Glucose

Identifieur interne : 002964 ( Main/Exploration ); précédent : 002963; suivant : 002965

Probing the Metabolic Network in Bloodstream-Form Trypanosoma brucei Using Untargeted Metabolomics with Stable Isotope Labelled Glucose

Auteurs : Darren J. Creek [Australie] ; Muriel Mazet [France] ; Fiona Achcar [Royaume-Uni] ; Jana Anderson [Royaume-Uni] ; Dong-Hyun Kim [Royaume-Uni] ; Ruwida Kamour [Libye] ; Pauline Morand [France] ; Yoann Millerioux [France] ; Marc Biran [France] ; Eduard J. Kerkhoven [Suède] ; Achuthanunni Chokkathukalam [Royaume-Uni] ; Stefan K. Weidt [Royaume-Uni] ; Karl E. V. Burgess [Royaume-Uni] ; Rainer Breitling [Royaume-Uni] ; David G. Watson [Royaume-Uni] ; Frédéric Bringaud [France] ; Michael P. Barrett [Royaume-Uni]

Source :

RBID : PMC:4361558

Descripteurs français

English descriptors

Abstract

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.


Url:
DOI: 10.1371/journal.ppat.1004689
PubMed: 25775470
PubMed Central: 4361558


Affiliations:


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Le document en format XML

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<country xml:lang="fr">France</country>
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<name sortKey="Millerioux, Yoann" sort="Millerioux, Yoann" uniqKey="Millerioux Y" first="Yoann" last="Millerioux">Yoann Millerioux</name>
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<addr-line>Centre de Résonance Magnétique des Systèmes Biologiques, Université de Bordeaux, CNRS UMR-5536, Bordeaux, France</addr-line>
</nlm:aff>
<country xml:lang="fr">France</country>
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<name sortKey="Biran, Marc" sort="Biran, Marc" uniqKey="Biran M" first="Marc" last="Biran">Marc Biran</name>
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<addr-line>Centre de Résonance Magnétique des Systèmes Biologiques, Université de Bordeaux, CNRS UMR-5536, Bordeaux, France</addr-line>
</nlm:aff>
<country xml:lang="fr">France</country>
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<region type="old region">Aquitaine</region>
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<settlement type="city">Bordeaux</settlement>
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<author>
<name sortKey="Kerkhoven, Eduard J" sort="Kerkhoven, Eduard J" uniqKey="Kerkhoven E" first="Eduard J." last="Kerkhoven">Eduard J. Kerkhoven</name>
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<nlm:aff id="aff007">
<addr-line>Systems and Synthetic Biology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Göteborg, Sweden</addr-line>
</nlm:aff>
<country xml:lang="fr">Suède</country>
<wicri:regionArea>Systems and Synthetic Biology, Department of Chemical and Biological Engineering, Chalmers University of Technology, Göteborg</wicri:regionArea>
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<author>
<name sortKey="Chokkathukalam, Achuthanunni" sort="Chokkathukalam, Achuthanunni" uniqKey="Chokkathukalam A" first="Achuthanunni" last="Chokkathukalam">Achuthanunni Chokkathukalam</name>
<affiliation wicri:level="4">
<nlm:aff id="aff008">
<addr-line>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow</wicri:regionArea>
<orgName type="university">Université de Glasgow</orgName>
<placeName>
<settlement type="city">Glasgow</settlement>
<region type="country">Écosse</region>
</placeName>
</affiliation>
</author>
<author>
<name sortKey="Weidt, Stefan K" sort="Weidt, Stefan K" uniqKey="Weidt S" first="Stefan K." last="Weidt">Stefan K. Weidt</name>
<affiliation wicri:level="4">
<nlm:aff id="aff008">
<addr-line>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow</wicri:regionArea>
<orgName type="university">Université de Glasgow</orgName>
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<settlement type="city">Glasgow</settlement>
<region type="country">Écosse</region>
</placeName>
</affiliation>
</author>
<author>
<name sortKey="Burgess, Karl E V" sort="Burgess, Karl E V" uniqKey="Burgess K" first="Karl E. V." last="Burgess">Karl E. V. Burgess</name>
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<nlm:aff id="aff008">
<addr-line>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow</wicri:regionArea>
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<settlement type="city">Glasgow</settlement>
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</author>
<author>
<name sortKey="Breitling, Rainer" sort="Breitling, Rainer" uniqKey="Breitling R" first="Rainer" last="Breitling">Rainer Breitling</name>
<affiliation wicri:level="4">
<nlm:aff id="aff009">
<addr-line>Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Manchester Institute of Biotechnology, Faculty of Life Sciences, University of Manchester, Manchester</wicri:regionArea>
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<settlement type="city">Manchester</settlement>
<region type="country">Angleterre</region>
<region type="région" nuts="1">Grand Manchester</region>
<settlement type="city">Manchester</settlement>
</placeName>
<orgName type="university">Université de Manchester</orgName>
</affiliation>
</author>
<author>
<name sortKey="Watson, David G" sort="Watson, David G" uniqKey="Watson D" first="David G." last="Watson">David G. Watson</name>
<affiliation wicri:level="1">
<nlm:aff id="aff010">
<addr-line>Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow</wicri:regionArea>
<wicri:noRegion>Glasgow</wicri:noRegion>
</affiliation>
</author>
<author>
<name sortKey="Bringaud, Frederic" sort="Bringaud, Frederic" uniqKey="Bringaud F" first="Frédéric" last="Bringaud">Frédéric Bringaud</name>
<affiliation wicri:level="4">
<nlm:aff id="aff002">
<addr-line>Centre de Résonance Magnétique des Systèmes Biologiques, Université de Bordeaux, CNRS UMR-5536, Bordeaux, France</addr-line>
</nlm:aff>
<country xml:lang="fr">France</country>
<wicri:regionArea>Centre de Résonance Magnétique des Systèmes Biologiques, Université de Bordeaux, CNRS UMR-5536, Bordeaux</wicri:regionArea>
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<region type="region">Nouvelle-Aquitaine</region>
<region type="old region">Aquitaine</region>
<settlement type="city">Bordeaux</settlement>
<settlement type="city">Bordeaux</settlement>
</placeName>
<orgName type="university">Université de Bordeaux</orgName>
</affiliation>
</author>
<author>
<name sortKey="Barrett, Michael P" sort="Barrett, Michael P" uniqKey="Barrett M" first="Michael P." last="Barrett">Michael P. Barrett</name>
<affiliation wicri:level="4">
<nlm:aff id="aff003">
<addr-line>Wellcome Trust Centre of Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Wellcome Trust Centre of Molecular Parasitology, Institute of Infection, Immunity and Inflammation, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow</wicri:regionArea>
<orgName type="university">Université de Glasgow</orgName>
<placeName>
<settlement type="city">Glasgow</settlement>
<region type="country">Écosse</region>
</placeName>
</affiliation>
<affiliation wicri:level="4">
<nlm:aff id="aff008">
<addr-line>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow, United Kingdom</addr-line>
</nlm:aff>
<country xml:lang="fr">Royaume-Uni</country>
<wicri:regionArea>Glasgow Polyomics, Wolfson Wohl Cancer Research Centre, Garscube Campus, College of Medical Veterinary & Life Sciences, University of Glasgow, Glasgow</wicri:regionArea>
<orgName type="university">Université de Glasgow</orgName>
<placeName>
<settlement type="city">Glasgow</settlement>
<region type="country">Écosse</region>
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</author>
</analytic>
<series>
<title level="j">PLoS Pathogens</title>
<idno type="ISSN">1553-7366</idno>
<idno type="eISSN">1553-7374</idno>
<imprint>
<date when="2015">2015</date>
</imprint>
</series>
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<textClass>
<keywords scheme="KwdEn" xml:lang="en">
<term>Animals</term>
<term>Cells, Cultured</term>
<term>Glucose (metabolism)</term>
<term>Glycerol (metabolism)</term>
<term>Metabolic Networks and Pathways (physiology)</term>
<term>Metabolomics (methods)</term>
<term>Oxidation-Reduction</term>
<term>Pentose Phosphate Pathway (physiology)</term>
<term>Succinic Acid (metabolism)</term>
<term>Trypanosoma brucei brucei (metabolism)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr">
<term>Acide succinique (métabolisme)</term>
<term>Animaux</term>
<term>Cellules cultivées</term>
<term>Glucose (métabolisme)</term>
<term>Glycérol (métabolisme)</term>
<term>Métabolomique ()</term>
<term>Oxydoréduction</term>
<term>Trypanosoma brucei brucei (métabolisme)</term>
<term>Voie des pentoses phosphates (physiologie)</term>
<term>Voies et réseaux métaboliques (physiologie)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en">
<term>Glucose</term>
<term>Glycerol</term>
<term>Succinic Acid</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en">
<term>Trypanosoma brucei brucei</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en">
<term>Metabolomics</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr">
<term>Acide succinique</term>
<term>Glucose</term>
<term>Glycérol</term>
<term>Trypanosoma brucei brucei</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr">
<term>Voie des pentoses phosphates</term>
<term>Voies et réseaux métaboliques</term>
</keywords>
<keywords scheme="MESH" qualifier="physiology" xml:lang="en">
<term>Metabolic Networks and Pathways</term>
<term>Pentose Phosphate Pathway</term>
</keywords>
<keywords scheme="MESH" xml:lang="en">
<term>Animals</term>
<term>Cells, Cultured</term>
<term>Oxidation-Reduction</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr">
<term>Animaux</term>
<term>Cellules cultivées</term>
<term>Métabolomique</term>
<term>Oxydoréduction</term>
</keywords>
</textClass>
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</teiHeader>
<front>
<div type="abstract" xml:lang="en">
<p>Metabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of
<italic>Trypanosoma brucei</italic>
, 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.</p>
</div>
</front>
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<name sortKey="Miranda Saavedra, D" uniqKey="Miranda Saavedra D">D Miranda-Saavedra</name>
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<name sortKey="Lamont, D" uniqKey="Lamont D">D Lamont</name>
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<name sortKey="Scheltema, R" uniqKey="Scheltema R">R Scheltema</name>
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<author>
<name sortKey="Breitling, R" uniqKey="Breitling R">R Breitling</name>
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<author>
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<name sortKey="Barrett, Mp" uniqKey="Barrett M">MP Barrett</name>
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<li>Suède</li>
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<li>Angleterre</li>
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<name sortKey="Achcar, Fiona" sort="Achcar, Fiona" uniqKey="Achcar F" first="Fiona" last="Achcar">Fiona Achcar</name>
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<name sortKey="Anderson, Jana" sort="Anderson, Jana" uniqKey="Anderson J" first="Jana" last="Anderson">Jana Anderson</name>
<name sortKey="Barrett, Michael P" sort="Barrett, Michael P" uniqKey="Barrett M" first="Michael P." last="Barrett">Michael P. Barrett</name>
<name sortKey="Barrett, Michael P" sort="Barrett, Michael P" uniqKey="Barrett M" first="Michael P." last="Barrett">Michael P. Barrett</name>
<name sortKey="Breitling, Rainer" sort="Breitling, Rainer" uniqKey="Breitling R" first="Rainer" last="Breitling">Rainer Breitling</name>
<name sortKey="Burgess, Karl E V" sort="Burgess, Karl E V" uniqKey="Burgess K" first="Karl E. V." last="Burgess">Karl E. V. Burgess</name>
<name sortKey="Chokkathukalam, Achuthanunni" sort="Chokkathukalam, Achuthanunni" uniqKey="Chokkathukalam A" first="Achuthanunni" last="Chokkathukalam">Achuthanunni Chokkathukalam</name>
<name sortKey="Kim, Dong Hyun" sort="Kim, Dong Hyun" uniqKey="Kim D" first="Dong-Hyun" last="Kim">Dong-Hyun Kim</name>
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</record>

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