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Journal of Pharmaceutical and... Jan 2021In T. cruzi, a causative agent of Chagas disease, phosphoenolpyruvate carboxykinase (TcPEPCK) is associated with carbohydrate catabolism. Due to its importance in the...
In T. cruzi, a causative agent of Chagas disease, phosphoenolpyruvate carboxykinase (TcPEPCK) is associated with carbohydrate catabolism. Due to its importance in the metabolism of the parasite, it has become a promising target for the development of new drugs against Chagas disease. Aiming to investigate different approaches for ligands screening, TcPEPCK was immobilized on amine-terminated magnetic beads (TcPEPCK-MB) and kinetically characterized by liquid chromatography tandem mass spectrometry activity assay with a K value of 10 ± 1 μM to oxaloacetate as substrate. Natural products library affords highly diverse molecular frameworks through their secondary metabolites, herein a ligand fishing TcPEPCK-MB assay is described for prospecting ligands in four ethanolic extracts of Brazilian Cerrado plants: Qualea grandiflora (Vochysiaceae), Diospyros burchellii (Ebenaceae), Anadenanthera falcata (Fabaceae) and Byrsonima coccolobifolia (Malpighiaceae). The chemical characterization of eleven identified ligands was carried out by liquid chromatography tandem high-resolution mass spectrometry experiments. Senecic acid, syneilesinolide A, phytosphingosine and vanillic acid 4-glucopyranoside are herein reported for the first time for Q. grandiflora, D. burchellii, A. falcata, respectively. In addition, the specificity of the assay was observed since only catechin was fished out from the ethanolic extract of B. coccolobifolia leaves, despite the presence of epicatechin epimer.
Topics: Brazil; Chagas Disease; Humans; Magnetic Phenomena; Phosphoenolpyruvate; Plant Extracts
PubMed: 33166842
DOI: 10.1016/j.jpba.2020.113710 -
Biotechnology Letters Feb 2017N-Acetyl-D-neuraminic acid (Neu5Ac) is often synthesized from exogenous N-acetylglucosamine (GlcNAc) and excess pyruvate. We have previously constructed a recombinant...
OBJECTIVES
N-Acetyl-D-neuraminic acid (Neu5Ac) is often synthesized from exogenous N-acetylglucosamine (GlcNAc) and excess pyruvate. We have previously constructed a recombinant Escherichia coli strain for Neu5Ac production using GlcNAc and intracellular phosphoenolpyruvate (PEP) as substrates (Zhu et al. Biotechnol Lett 38:1-9, 2016).
RESULTS
PEP synthesis-related genes, pck and ppsA, were overexpressed within different modes to construct PEP-supply modules, and their effects on Neu5Ac production were investigated. All the PEP-supply modules enhanced Neu5Ac production. For the best module, pCDF-pck-ppsA increased Neu5Ac production to 8.6 ± 0.15 g l, compared with 3.6 ± 0.15 g l of the original strain. Neu5Ac production was further increased to 15 ± 0.33 g l in a 1 l fermenter.
CONCLUSIONS
The PEP-supply module can improve the intracellular PEP supply and enhance Neu5Ac production, which benefited industrial Neu5Ac production.
Topics: Biocatalysis; Escherichia coli; Phosphoenolpyruvate; Pyruvic Acid
PubMed: 27738778
DOI: 10.1007/s10529-016-2235-8 -
Chemistry & Biology Feb 1996The diverse enzymes that use phosphoenolpyruvate as a substrate lie at the heart of cellular energy metabolism, as well as a number of critical biosynthetic pathways.... (Review)
Review
The diverse enzymes that use phosphoenolpyruvate as a substrate lie at the heart of cellular energy metabolism, as well as a number of critical biosynthetic pathways. The versatility of the enol ether linkage is reflected not only in the rich chemistry and enzymology of PEP, but also in the variety of metabolites in which the high-energy enol ether linkage is preserved.
Topics: Energy Metabolism; Enzymes; Molecular Probes; Phosphoenolpyruvate; Polysaccharides; Substrate Specificity; Vinyl Compounds
PubMed: 8807832
DOI: 10.1016/s1074-5521(96)90282-3 -
European Journal of Cancer (Oxford,... Jan 2021Metabolic enzymes have non-canonical functions and play vital roles in the regulation of various cellular activities. Phosphoenolpyruvate carboxykinase 1 (PCK1), a...
Association of phosphoenolpyruvate carboxykinase 1 protein kinase activity-dependent sterol regulatory element-binding protein 1 activation with prognosis of oesophageal carcinoma.
BACKGROUND
Metabolic enzymes have non-canonical functions and play vital roles in the regulation of various cellular activities. Phosphoenolpyruvate carboxykinase 1 (PCK1), a gluconeogenic enzyme, was recently identified as an AKT-dependent protein kinase and promoted sterol regulatory element-binding protein 1 (SREBP1)-dependent lipogenesis. However, association of this protein kinase activity of PCK1 with progression of oesophageal squamous cell carcinoma (ESCC) is unclear.
METHODS
We examined 200 ESCC patient samples and prognosis using immunohistochemistry, multivariate Cox regression and Kaplan-Meier Plot analyses.
RESULTS
We show that the expression levels of AKT pS473, AKT-regulated PCK1 pS90, PCK1-mediated INSIG1 pS207/INSIG2 pS151 and nuclear SREBP1 were higher in analysed 200 human ESCC specimens than in their adjacent non-tumour tissues; the expression levels of these proteins were significantly and positively correlated with each other in tumour specimens. In addition, the expression levels of PCK1 pS90, INSIG1 pS207/INSIG2 pS151 and SREBP1 were associated with the tumour, node and metastasis stage and progression in ESCC. Importantly, levels of PCK1 pS90 or INSIG1 pS207/INSIG2 pS151 or nuclear SREBP1 were positively correlated with poor prognosis in patients with ESCC, and the combined expression values of PCK1 pS90, INSIG1 pS207/INSIG2 pS151 and nuclear SREBP1 had a better prognostic value than that of each individual protein expression value and was an independent prognostic marker for ESCC.
CONCLUSION
These findings reveal the role of PCK1 protein kinase activity-dependent SREBP1 activation in ESCC progression. The regulation of SREBP1 by AKT activation-dependent PCK1 protein kinase activity may provide the potential for the diagnosis and treatment of human ESCC.
Topics: Esophageal Neoplasms; Female; Humans; Male; Middle Aged; Phosphoenolpyruvate; Prognosis; Retrospective Studies; Sterol Regulatory Element Binding Protein 1
PubMed: 33278777
DOI: 10.1016/j.ejca.2020.09.040 -
The New Phytologist Aug 2012
Topics: Carbon; Cell Respiration; Mitochondria; Oxidative Stress; Ozone; Phosphoenolpyruvate; Phosphoenolpyruvate Carboxylase; Plant Leaves; Plants
PubMed: 22686461
DOI: 10.1111/j.1469-8137.2012.04211.x -
The structure of Synechococcus elongatus enolase reveals key aspects of phosphoenolpyruvate binding.Acta Crystallographica. Section F,... Apr 2022A structure-function characterization of Synechococcus elongatus enolase (SeEN) is presented, representing the first structural report on a cyanobacterial enolase. X-ray...
A structure-function characterization of Synechococcus elongatus enolase (SeEN) is presented, representing the first structural report on a cyanobacterial enolase. X-ray crystal structures of SeEN in its apoenzyme form and in complex with phosphoenolpyruvate are reported at 2.05 and 2.30 Å resolution, respectively. SeEN displays the typical fold of enolases, with a conformationally flexible loop that closes the active site upon substrate binding, assisted by two metal ions that stabilize the negatively charged groups. The enzyme exhibits a catalytic efficiency of 1.2 × 10 M s for the dehydration of 2-phospho-D-glycerate, which is comparable to the kinetic parameters of related enzymes. These results expand the understanding of the biophysical features of these enzymes, broadening the toolbox for metabolic engineering applications.
Topics: Crystallography, X-Ray; Phosphoenolpyruvate; Phosphopyruvate Hydratase; Synechococcus
PubMed: 35400670
DOI: 10.1107/S2053230X22003612 -
Biochimica Et Biophysica Acta Apr 2007We report here initial studies on phosphoenolpyruvate metabolism in coupled mitochondria isolated from Jerusalem artichoke tubers. It was found that: (1)...
We report here initial studies on phosphoenolpyruvate metabolism in coupled mitochondria isolated from Jerusalem artichoke tubers. It was found that: (1) phosphoenolpyruvate can be metabolized by Jerusalem artichoke mitochondria by virtue of the presence of the mitochondrial pyruvate kinase, shown both immunologically and functionally, located in the inner mitochondrial compartments and distinct from the cytosolic pyruvate kinase as shown by the different pH and inhibition profiles. (2) Jerusalem artichoke mitochondria can take up externally added phosphoenolpyruvate in a proton compensated manner, in a carrier-mediated process which was investigated by measuring fluorimetrically the oxidation of intramitochondrial pyridine nucleotide which occurs as a result of phosphoenolpyruvate uptake and alternative oxidase activation. (3) The addition of phosphoenolpyruvate causes pyruvate and ATP production, as monitored via HPLC, with their efflux into the extramitochondrial phase investigated fluorimetrically. Such an efflux occurs via the putative phosphoenolpyruvate/pyruvate and phosphoenolpyruvate/ATP antiporters, which differ from each other and from the pyruvate and the adenine nucleotide carriers, in the light of the different sensitivity to non-penetrant compounds. These carriers were shown to regulate the rate of efflux of both pyruvate and ATP. The appearance of citrate and oxaloacetate outside mitochondria was also found as a result of phosphoenolpyruvate addition.
Topics: Adenosine Triphosphate; Biological Transport; Helianthus; Mitochondria; Phosphoenolpyruvate; Pyruvate Kinase; Pyruvic Acid
PubMed: 17418088
DOI: 10.1016/j.bbabio.2007.02.010 -
Sheng Wu Gong Cheng Xue Bao = Chinese... Dec 2011Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is an important ubiquitous cytosol enzyme that fixes HCO3 together with phosphoenolpyruvate (PEP) and yields... (Review)
Review
Phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) is an important ubiquitous cytosol enzyme that fixes HCO3 together with phosphoenolpyruvate (PEP) and yields oxaloacetate that can be converted to intermediates of the citric acid cycle. In plant cells, PEPC participates in CO2 assimilation and other important metabolic pathways, and it has broad functions in different plant tissues. PEPC is also involved in the regulation of storage product synthesis and metabolism in seeds, such as affecting the metabolic fluxes from sugars/starch towards the synthesis of fatty acids or amino acids and proteins. In this review, we introduced the progress in classification, structure and regulation of PEPC in plant tissues. We discussed the potential applications of plant PEPCs in genetic engineering. The researches in functions and regulation mechanism of plant PEPCs will provide beneficial approaches to applications of plant PEPCs in high-yield crops breeding, energy crop and microbe genetic engineering.
Topics: Bicarbonates; Genetic Engineering; Oxaloacetic Acid; Phosphoenolpyruvate; Phosphoenolpyruvate Carboxylase; Plants
PubMed: 22506410
DOI: No ID Found -
Science China. Life Sciences Mar 2023
Topics: Nitrogen Fixation; Phosphoenolpyruvate; Root Nodules, Plant; Symbiosis; Nitrogen
PubMed: 36515862
DOI: 10.1007/s11427-022-2256-2 -
Biochimica Et Biophysica Acta Dec 1981(1) The relationship between phosphoenolpyruvate formation and its accumulation in kidney cortex mitochondria of rabbit was studied in the presence of glutamate as...
(1) The relationship between phosphoenolpyruvate formation and its accumulation in kidney cortex mitochondria of rabbit was studied in the presence of glutamate as substrate. (2) In mitochondria incubated in either State 4 or under uncoupled conditions, both 1,2,3-benzenetricarboxylate and atractyloside resulted in a marked elevation of the intramitochondrial phosphoenolpyruvate accompanied by a 2-4-fold decline in production of this compound. The same effect was induced by n-butylmalonate in uncoupled mitochondria, while both phosphoenolpyruvate efflux and its production were inhibited to a smaller extent in mitochondria incubated with 1,2,3-benzenetricarboxylate in State 3. (3) Citrate, malate or 2-phosphoglycerate caused a fast displacement of phosphoenolpyruvate from atractyloside-inhibited mitochondria to the reaction medium. In contrast, on the addition of ATP to mitochondria incubated with 1,2,3-benzenetricarboxylate, the rate of phosphoenolpyruvate efflux was lower than that induced by either malate or citrate. (4) Despite the presence of both 1,2,3-benzenetricarboxylate and atractyloside, arsenite and rotenone plus antimycin resulted in a leakage of phosphoenolpyruvate from the mitochondria, probably via a carrier-independent mechanism. (5) Based on the present results it seems that depending on the metabolic condition, the tricarboxylate carrier and the adenine nucleotide translocase are functioning to different extents in the efflux of phosphoenolpyruvate from rabbit renal mitochondria to the surrounding medium.
Topics: Animals; Atractyloside; Benzene Derivatives; Kidney Cortex; Kinetics; Male; Mitochondria; Phosphoenolpyruvate; Rabbits; Rotenone; Tricarboxylic Acids
PubMed: 7317387
DOI: 10.1016/0005-2728(81)90234-6