2014
DOI: 10.1074/jbc.m114.567230
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Contribution of Pyruvate Phosphate Dikinase in the Maintenance of the Glycosomal ATP/ADP Balance in the Trypanosoma brucei Procyclic Form

Abstract: Background:The role of pyruvate phosphate dikinase (PPDK), which catalyzes a reversible reaction, is unknown in many eukaryotes. Results: Deletion of the trypanosomal PPDK gene affects glycolysis. Conclusion:In trypanosomes, PPDK works in the glycolytic direction and participates in the maintenance of the glycosomal ATP/ADP balance. Significance: The glycosomal PPDK provides a metabolic flexibility by producing 2 ATP per phosphoenolpyruvate consumed.

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Cited by 39 publications
(56 citation statements)
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“…Characterization of the FRDg gene closed a long-standing debate about the existence of bona fide soluble NADH-dependent FRD in trypanosomes Turrens, 1999) and strengthened Gest's hypothesis, as glucose metabolism in the trypanosome glycosomes closely resembles his succinic fermentation model. Indeed, inclusion of the glycosomal succinate branch in the procyclic trypanosomes releases PEP molecules used to produce ATP in the cytosol through pyruvate kinase activity, which has been described as essential for the viability of the procyclic form (Morris et al, 2002;Coustou et al, 2003;Deramchia et al, 2014). The trypanosome FRD enzymes were the first soluble NADH-dependent FRD characterized, although the concept might be more widely applicable as soluble NADH-dependent FRD activity was reported in the parasite Trichomonas foetus (Müller, 2003) and the prokaryote Hydrogenobacter thermophilus (Miura et al, 2008).…”
Section: Succinic Fermentationmentioning
confidence: 99%
See 1 more Smart Citation
“…Characterization of the FRDg gene closed a long-standing debate about the existence of bona fide soluble NADH-dependent FRD in trypanosomes Turrens, 1999) and strengthened Gest's hypothesis, as glucose metabolism in the trypanosome glycosomes closely resembles his succinic fermentation model. Indeed, inclusion of the glycosomal succinate branch in the procyclic trypanosomes releases PEP molecules used to produce ATP in the cytosol through pyruvate kinase activity, which has been described as essential for the viability of the procyclic form (Morris et al, 2002;Coustou et al, 2003;Deramchia et al, 2014). The trypanosome FRD enzymes were the first soluble NADH-dependent FRD characterized, although the concept might be more widely applicable as soluble NADH-dependent FRD activity was reported in the parasite Trichomonas foetus (Müller, 2003) and the prokaryote Hydrogenobacter thermophilus (Miura et al, 2008).…”
Section: Succinic Fermentationmentioning
confidence: 99%
“…pyruvate phosphate dikinase (PPDK, step 5) working in the gluconeogenic direction (Bringaud et al, 1998), PEPCK (step 1) and malate dehydrogenase (step 2), may operate without any impact on the glycosomal ATP/ADP balance as ATP molecules consumed by PPDK are regenerated by PEPCK. Its utilization is certainly exceptional, as the procyclic cells growing in standard glucose-rich conditions preferentially use PPDK in the glycolytic direction to participate in the maintenance of this glycosomal ATP/ADP balance (Deramchia et al, 2014). The relevance of this temporarily used hypothetical cycle is its ability to produce cytosolic NADPH sustained by NADH production in the glycosomes, which may compensate for the absence of cytosolic (and mitochondrial) transhydrogenases in trypanosomatids.…”
Section: Connections Between the Acetate And Succinate Branches Provimentioning
confidence: 99%
“…In conclusion, we describe here a new mechanism for the regulation of nutrient utilization based on the competition between two enzymes (kinases) for a common substrate (ATP). The sequestration of the two kinases in the glycosomes is key to this mechanism since these peroxisome-like organelles show limited or no nucleotide exchange with the cytosol at a metabolic timescale (Deramchia et al, 2014; Haanstra et al, 2014). Hence, the ATP pool available to glycosomal kinases is limited, offering a situation where significant excess of one kinase (here GK) can abolish almost completely the flux through the other one (HK).…”
Section: Resultsmentioning
confidence: 99%
“…To investigate the biological role of TbSLP in the procyclic form of T. brucei , the TbSLP gene was deleted by gene replacement as previously described . Replacement of both TbSLP alleles by the puromycin and blasticidin selectable markers in the procyclic EATRO1125.T7T background was confirmed by PCR (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…Replacement of the TbSLP gene by the blasticidin and puromycin resistance markers via homologous recombination was performed with DNA fragments containing the resistance marker gene flanked by TbSLP UTR sequences, as described previously . Briefly, the pGEMt plasmid was used to clone a HpaI DNA fragment containing either the blasticidin or puromycin resistance marker gene preceded by the TbSLP 5'‐UTR fragment (758 bp) and followed by the TbSLP 3'‐UTR fragment (754 bp).…”
Section: Methodsmentioning
confidence: 99%