The glycosomal ATP-dependent phosphofructokinase of Trypanosoma brucei operates also in the gluconeogenic direction.

IF 9.8 1区 生物学 Q1 Agricultural and Biological Sciences
PLoS Biology Pub Date : 2025-05-16 eCollection Date: 2025-05-01 DOI:10.1371/journal.pbio.3002938
Nicolas Plazolles, Hanna Kulyk, Edern Cahoreau, Marc Biran, Marion Wargnies, Erika Pineda, Mohammad El Kadri, Aline Rimoldi, Perrine Hervé, Corinne Asencio, Loïc Rivière, Paul A M Michels, Daniel Inaoka, Emmanuel Tetaud, Jean-Charles Portais, Frédéric Bringaud
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引用次数: 0

Abstract

In the glucose-free environment of the midgut of the tsetse fly vector, the procyclic forms of Trypanosoma brucei primarily consume proline to feed its central carbon and energy metabolism. In this context, the parasite produces through gluconeogenesis, glucose 6-phosphate (G6P), the precursor of essential metabolic pathways, from proline catabolism. We show here that the parasite uses three different enzymes to perform the key gluconeogenic reaction producing fructose 6-phosphate (F6P) from fructose 1,6-bisphosphate, (i) fructose-1,6-bisphosphatase (FBPase), the canonical enzyme performing this reaction, (ii) sedoheptulose-1,7-bisphosphatase (SBPase), and (iii) more surprisingly ATP-dependent phosphofructokinase (PFK), an enzyme considered to irreversibly catalyze the opposite reaction involved in glycolysis. These three enzymes, as well as six other glycolytic/gluconeogenic enzymes, are located in peroxisome-related organelles, named glycosomes. Incorporation of 13C-enriched glycerol (a more effective alternative to proline for monitoring gluconeogenic activity) into F6P and G6P was more affected in the PFK null mutant than in the FBPase null mutant, suggesting the PFK contributes at least as much as FBPase to gluconeogenesis. We also showed that glucose deprivation did not affect the quantities of PFK substrates and products, whereas an approximately 500-fold increase in the substrate/product ratio was expected for PFK to carry out the gluconeogenic reaction. In conclusion, we show for the first time that ATP-dependent PFK can function in vivo in the gluconeogenic direction, even in the presence of FBPase activity. This particular feature, which precludes loss of ATP through a futile cycle involving PFK and FBPase working simultaneously in the glycolytic and gluconeogenic directions, respectively, is possibly due to the supramolecular organization of the metabolic pathway within glycosomes to overcome thermodynamic barriers through metabolic channeling.

布鲁氏锥虫的糖体atp依赖性磷酸果糖激酶也在糖异生方向起作用。
在采采蝇载体中肠的无糖环境中,布氏锥虫的原环型主要消耗脯氨酸来喂养其中心碳和能量代谢。在这种情况下,寄生虫通过糖异生产生葡萄糖6-磷酸(G6P),这是脯氨酸分解代谢的基本代谢途径的前体。我们在这里表明,寄生虫使用三种不同的酶来完成关键的糖异生反应,从果糖1,6-二磷酸产生果糖6-磷酸(F6P), (i)果糖-1,6-二磷酸酶(FBPase),执行该反应的典型酶,(ii) sedoheptulose-1,7-二磷酸酶(SBPase),以及(iii)更令人惊讶的atp依赖性磷酸果糖激酶(PFK),一种酶被认为是不可逆地催化糖酵解的相反反应。这三种酶,以及其他六种糖酵解/糖异生酶,位于过氧化物酶体相关的细胞器中,称为糖体。在F6P和G6P中掺入富含13c的甘油(一种比脯氨酸更有效的监测糖异生活性的替代品)在PFK无突变体中比在FBPase无突变体中受到更大的影响,这表明PFK对糖异生的贡献至少与FBPase一样多。我们还发现,葡萄糖剥夺不会影响PFK底物和产物的数量,而预计PFK进行糖异生反应的底物/产物比增加约500倍。总之,我们首次证明atp依赖性PFK在体内即使存在FBPase活性也能在糖异生方向上起作用。PFK和FBPase分别在糖酵解和糖异生方向上同时工作,这一特殊的特征阻止了ATP通过无效循环的损失,这可能是由于糖体内代谢途径的超分子组织通过代谢通道克服热力学障碍。
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来源期刊
PLoS Biology
PLoS Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-BIOLOGY
CiteScore
15.40
自引率
2.00%
发文量
359
审稿时长
3-8 weeks
期刊介绍: PLOS Biology is the flagship journal of the Public Library of Science (PLOS) and focuses on publishing groundbreaking and relevant research in all areas of biological science. The journal features works at various scales, ranging from molecules to ecosystems, and also encourages interdisciplinary studies. PLOS Biology publishes articles that demonstrate exceptional significance, originality, and relevance, with a high standard of scientific rigor in methodology, reporting, and conclusions. The journal aims to advance science and serve the research community by transforming research communication to align with the research process. It offers evolving article types and policies that empower authors to share the complete story behind their scientific findings with a diverse global audience of researchers, educators, policymakers, patient advocacy groups, and the general public. PLOS Biology, along with other PLOS journals, is widely indexed by major services such as Crossref, Dimensions, DOAJ, Google Scholar, PubMed, PubMed Central, Scopus, and Web of Science. Additionally, PLOS Biology is indexed by various other services including AGRICOLA, Biological Abstracts, BIOSYS Previews, CABI CAB Abstracts, CABI Global Health, CAPES, CAS, CNKI, Embase, Journal Guide, MEDLINE, and Zoological Record, ensuring that the research content is easily accessible and discoverable by a wide range of audiences.
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