158 results on '"glycolytic enzyme"'
Search Results
52. Biochemical characterisation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from the liver fluke, Fasciola hepatica.
- Author
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Zinsser, Veronika L., Hoey, Elizabeth M., Trudgett, Alan, and Timson, David J.
- Subjects
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GLYCERALDEHYDEPHOSPHATE dehydrogenase , *FASCIOLA hepatica , *LIVER flukes , *NICOTINAMIDE adenine dinucleotide phosphate , *HOMODIMERS , *GLYCOLYSIS , *ESCHERICHIA coli proteins - Abstract
Abstract: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) catalyses one of the two steps in glycolysis which generate the reduced coenzyme NADH. This reaction precedes the two ATP generating steps. Thus, inhibition of GAPDH will lead to substantially reduced energy generation. Consequently, there has been considerable interest in developing GAPDH inhibitors as anti-cancer and anti-parasitic agents. Here, we describe the biochemical characterisation of GAPDH from the common liver fluke Fasciola hepatica (FhGAPDH). The primary sequence of FhGAPDH is similar to that from other trematodes and the predicted structure shows high similarity to those from other animals including the mammalian hosts. FhGAPDH lacks a binding pocket which has been exploited in the design of novel antitrypanosomal compounds. The protein can be expressed in, and purified from Escherichia coli; the recombinant protein was active and showed no cooperativity towards glyceraldehyde 3-phosphate as a substrate. In the absence of ligands, FhGAPDH was a mixture of homodimers and tetramers, as judged by protein–protein crosslinking and analytical gel filtration. The addition of either NAD+ or glyceraldehyde 3-phosphate shifted this equilibrium towards a compact dimer. Thermal scanning fluorimetry demonstrated that this form was considerably more stable than the unliganded one. These responses to ligand binding differ from those seen in mammalian enzymes. These differences could be exploited in the discovery of reagents which selectively disrupt the function of FhGAPDH. [Copyright &y& Elsevier]
- Published
- 2014
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53. Identification of Alternative Allosteric Sites in Glycolytic Enzymes for Potential Use as Species-Specific Drug Targets
- Author
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Serkan Celiker, Fatih Ozhelvaci, E. Demet Akten, Merve Ayyildiz, Akten, Ebru Demet, and Ayyıldız, Merve
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0301 basic medicine ,Drug ,media_common.quotation_subject ,Allosteric regulation ,Dehydrogenase ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Biochemistry ,drug discovery ,03 medical and health sciences ,0302 clinical medicine ,elastic network modeling ,Molecular Biosciences ,Binding site ,Receptor ,Molecular Biology ,lcsh:QH301-705.5 ,media_common ,Original Research ,Chemistry ,Drug discovery ,glycolytic enzyme ,allosteric regulation ,species-specific ,030104 developmental biology ,lcsh:Biology (General) ,030220 oncology & carcinogenesis ,Pyruvate kinase ,Phosphofructokinase - Abstract
Three allosteric glycolytic enzymes, phosphofructokinase, glyceraldehyde-3 phosphate dehydrogenase and pyruvate kinase, associated with bacterial, parasitic and human species, were explored to identify potential allosteric sites that would be used as prime targets for species-specific drug design purposes using a newly developed approach which incorporates solvent mapping, elastic network modeling, sequence and structural alignments. The majority of binding sites detected by solvent mapping overlapped with the interface regions connecting the subunits, thus appeared as promising target sites for allosteric regulation. Each binding site was then evaluated by its ability to alter the global dynamics of the receptor defined by the percentage change in the frequencies of the lowest-frequency modes most significantly and as anticipated, the most effective ones were detected in the vicinity of the well-reported catalytic and allosteric sites. Furthermore, some of our proposed regions intersected with experimentally resolved sites which are known to be critical for activity regulation, which further validated our approach. Despite the high degree of structural conservation encountered between bacterial/parasitic and human glycolytic enzymes, the majority of the newly presented allosteric sites exhibited a low degree of sequence conservation which further increased their likelihood to be used as species-specific target regions for drug design studies. Tubitak
- Published
- 2020
54. Therapeutic targeting of cancer metabolism with triosephosphate isomerase
- Author
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Bursa Uludağ Üniversitesi/Fen-Edebiyat Fakültesi/Biyoloji Bölümü., Pekel, Gonca, and Arı, Ferda
- Subjects
Identification ,Pyruvate Kinase Deficiency of Red Cells ,Triose-Phosphate Isomerase ,Hereditary Hemolytic Anemia ,Biochemistry & molecular biology ,Carcinogenesis ,Triose-phosphate isomerase ,Proteomic analysis ,Glycolytic enzyme ,Pentose phosphate cycle ,Review ,Molecular model ,Metastasis ,Differential expression ,Chemical structure ,Neoplasms ,Antineoplastic agents ,Pathology ,Animals ,Humans ,Tumor marker ,Triosephosphate isomerase ,Breast-cancer ,Cancer ,Cell carcinoma ,Animal ,Gluconeogenesis ,Proteins ,Enzyme catalysis ,Enzyme inhibitors ,Enzyme inhibitor ,Fatty acid ,Nonhuman ,Chemistry, multidisciplinary ,Malignant neoplasm ,Chemistry ,Enzyme inhibition ,Metabolism ,Phenotype ,Crystal-structure ,Genes ,Antineoplastic agent ,Fatty acid synthesis ,Aerobic glycolysis ,Neoplasm ,Warburg effect ,Glycolysis ,Molecular structure ,Models, molecular ,Human - Abstract
The increase in glycolytic flux in cancer, known as aerobic glycolysis, is one of the most important hallmarks of cancer. Therefore, glycolytic enzymes have importance in understanding the molecular mechanism of cancer progression. Triosephosphate isomerase (TPI) is one of the key glycolytic enzymes. Furthermore, it takes a part in gluconeogenesis, pentose phosphate pathway and fatty acid biosynthesis. To date, it has been shown altered levels of TPI in various cancer types, especially in metastatic phenotype. According to other studies, TPI might be considered as a potential therapeutic target and a cancer-related biomarker in different types of cancer. However, its function in tumor formation and development has not been fully understood. Here, we reviewed the relationship between TPI and cancer for the first time.
- Published
- 2020
55. Triose phosphate isomerase from the blood fluke Schistosoma mansoni: Biochemical characterisation of a potential drug and vaccine target.
- Author
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Zinsser, Veronika L., Farnell, Edward, Dunne, David W., and Timson, David J.
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TRIOSE phosphates , *ISOMERASES , *SCHISTOSOMA mansoni , *DRUG synergism , *TARGETED drug delivery , *TEMPERATURE effect - Abstract
Highlights: [•] Schistosoma mansoni triose phosphate isomerase (SmTPI) is a dimer. [•] A Ser-Ala-Asp motif, believed to be a helminth-specific epitope, is surface-exposed. [•] The melting temperature of SmTPI is unusually high (82.0°C). [•] K m(dihydroxyacetone phosphate)=0.51mM; K m(glyceraldehyde 3-phosphate)=1.1mM. [•] Reagents which disrupt dimerization or modify Cys-221 may be effective inhibitors. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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56. Research progress on the interaction between long non‑coding RNAs and RNA‑binding proteins to influence the reprogramming of tumor glucose metabolism (Review).
- Author
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Wu W and Wen K
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- Glucose metabolism, Glycolysis, Humans, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, Neoplasms pathology, RNA, Long Noncoding genetics, RNA, Long Noncoding metabolism
- Abstract
As epigenetic regulators, long non‑coding RNAs (lncRNAs) are involved in various important regulatory processes and typically interact with RNA‑binding proteins (RBPs) to exert their core functional effects. An increasing number of studies have demonstrated that lncRNAs can regulate the occurrence and development of cancer through a variety of complex mechanisms and can also participate in tumor glucose metabolism by directly or indirectly regulating the Warburg effect. As one of the metabolic characteristics of tumor cells, the Warburg effect provides a large amount of energy and numerous intermediate products to meet the consumption demands of tumor metabolism, providing advantages for the occurrence and development of tumors. The present review article summarizes the regulatory effects of lncRNAs on the reprogramming of glucose metabolism after interacting with RBPs in tumors. The findings discussed herein may aid in the better understanding of the pathogenesis of malignancies, and may provide novel therapeutic targets, as well as new diagnostic and prognostic markers for human cancers.
- Published
- 2022
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57. Effects of dietary energy sources on early postmortem muscle metabolism of finishing pigs
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Feng Gao, Changning Yu, Lin Zhang, Jiaolong Li, Yanjiao Li, and Guanghong Zhou
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0301 basic medicine ,medicine.medical_specialty ,Starch ,Lactate dehydrogenase A ,lcsh:Animal biochemistry ,Carbohydrate metabolism ,Creatine ,Article ,Phosphocreatine ,03 medical and health sciences ,chemistry.chemical_compound ,Animal science ,Animal Products ,Internal medicine ,medicine ,education ,lcsh:QP501-801 ,lcsh:SF1-1100 ,education.field_of_study ,biology ,0402 animal and dairy science ,Finishing Pigs ,04 agricultural and veterinary sciences ,040201 dairy & animal science ,Postmortem Metabolism ,Glycolytic Enzyme ,030104 developmental biology ,Endocrinology ,chemistry ,biology.protein ,Animal Science and Zoology ,Creatine kinase ,lcsh:Animal culture ,Pyruvic acid ,PI3K/AKT-HIF-1α Pathway ,Energy source ,Dietary Energy Source ,Food Science - Abstract
Objective This study investigated the effects of different dietary energy sources on early postmortem muscle metabolism of finishing pigs. Methods Seventy-two barrow (Duroc×Landrace×Yorkshire, DLY) pigs (65.0±2.0 kg) were allotted to three iso-energetic and iso-nitrogenous diets: A (44.1% starch, 5.9% crude fat, and 12.6% neutral detergent fibre [NDF]), B (37.6% starch, 9.5% crude fat, and 15.4% NDF) or C (30.9% starch, 14.3% crude fat, and 17.8% NDF). After the duration of 28-day feeding experiment, 24 pigs (eight per treatment) were slaughtered and the M. longissimus lumborum (LL) samples at 45 min postmortem were collected. Results Compared with diet A, diet C resulted in greater adenosine triphosphate and decreased phosphocreatine (PCr) concentrations, greater activity of creatine kinase and reduced percentage bound activities of hexokinase (HK), and pyruvate kinase (PK) in LL muscles (p
- Published
- 2017
58. Implication of Potential Differential Roles of the Two Phosphoglucomutase Isoforms in the Protozoan Parasite Cryptosporidium parvum.
- Author
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Nie, Jiawen, Yin, Jigang, Wang, Dongqiang, Wang, Chenchen, and Zhu, Guan
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CRYPTOSPORIDIUM parvum ,CELL membranes ,PARASITES ,PROTOZOA ,CRYPTOSPORIDIUM ,RECOMBINANT proteins ,INTRACELLULAR pathogens - Abstract
Phosphoglucomutase 1 (PGM1) catalyzes the conversion between glucose-1-phosphate and glucose-6-phosphate in the glycolysis/glucogenesis pathway. PGM1s are typically cytosolic enzymes in organisms lacking chloroplasts. However, the protozoan Cryptosporidium parasites possess two tandemly duplicated PGM1 genes evolved by a gene duplication after their split from other apicomplexans. Moreover, the downstream PGM1 isoform contains an N-terminal signal peptide, predicting a non-cytosolic location. Here we expressed recombinant proteins of the two PGM1 isoforms from the zoonotic Cryptosporidium parvum, namely CpPGM1A and CpPGM1B, and confirmed their enzyme activity. Both isoforms followed Michaelis–Menten kinetics towards glucose-1-phosphate (K
m = 0.17 and 0.13 mM, Vmax = 7.30 and 2.76 μmol/min/mg, respectively). CpPGM1A and CpPGM1B genes were expressed in oocysts, sporozoites and intracellular parasites at a similar pattern of expression, however CpPGM1A was expressed at much higher levels than CpPGM1B. Immunofluorescence assay showed that CpPGM1A was present in the cytosol of sporozoites, however this was enriched towards the plasma membranes in the intracellular parasites; whereas CpPGM1B was mainly present under sporozoite pellicle, although relocated to the parasitophorous vacuole membrane in the intracellular development. These observations indicated that CpPGM1A played a house-keeping function, while CpPGM1B played a different biological role that remains to be defined by future investigations. [ABSTRACT FROM AUTHOR]- Published
- 2022
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59. Reactions upstream of glycerate-1,3-bisphosphate drive Corynebacterium glutamicum d-lactate productivity under oxygen deprivation.
- Author
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Tsuge, Yota, Yamamoto, Shougo, Suda, Masako, Inui, Masayuki, and Yukawa, Hideaki
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CORYNEBACTERIUM , *OXYGEN , *LACTOBACILLUS delbrueckii , *BIODEGRADABLE plastics , *GENES - Abstract
We previously demonstrated the simplicity of oxygen-deprived Corynebacterium glutamicum to produce d-lactate, a primary building block of next-generation biodegradable plastics, at very high optical purity by introducing heterologous D- ldhA gene from Lactobacillus delbrueckii. Here, we independently evaluated the effects of overexpressing each of genes encoding the ten glycolytic enzymes on d-lactate production in C. glutamicum. We consequently show that while the reactions catalyzed by glucokinase (GLK), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), phosphofructokinase (PFK), triosephosphate isomerase (TPI), and bisphosphate aldolase had positive effects on d-lactate productivity by increasing 98, 39, 15, 13, and 10 %, respectively, in 10 h reactions in minimal salts medium, the reaction catalyzed by pyruvate kinase had large negative effect by decreasing 70 %. The other glycolytic enzymes did not affect d-lactate productivity when each of encoding genes was overexpressed. It is noteworthy that all reactions associated with positive effects are located upstream of glycerate-1,3-bisphosphate in the glycolytic pathway. The d-lactate yield also increased by especially overexpressing TPI encoding gene up to 94.5 %. Interestingly, overexpression of PFK encoding gene reduced the yield of succinate, one of the main by-products of d-lactate production, by 52 %, whereas overexpression of GAPDH encoding gene increased succinate yield by 26 %. Overexpression of GLK encoding gene markedly increased the yield of dihydroxyacetone and glycerol by 10- and 5.8-fold in exchange with decreasing the d-lactate yield. The effect of overexpressing glycolytic genes was also evaluated in 80 h long-term reactions. The variety of effects of overexpressing each of genes encoding the ten glycolytic enzymes on d-lactate production is discussed. [ABSTRACT FROM AUTHOR]
- Published
- 2013
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60. 3,4,5-tri- O-caffeoylquinic acid inhibits amyloid β-mediated cellular toxicity on SH-SY5Y cells through the upregulation of PGAM1 and G3PDH.
- Author
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Miyamae, Yusaku, Han, Junkyu, Sasaki, Kazunori, Terakawa, Mika, Isoda, Hiroko, and Shigemori, Hideyuki
- Abstract
Caffeoylquinic acid (CQA) is one of the phenylpropanoids found in a variety of natural resources and foods, such as sweet potatoes, propolis, and coffee. Previously, we reported that 3,5-di- O-caffeoylquinic acid (3,5-di-CQA) has a neuroprotective effect against amyloid-β (Aβ)-induced cell death through the overexpression of glycolytic enzyme. Additionally, 3,5-di-CQA administration induced the improvement of spatial learning and memory on senescence accelerated-prone mice (SAMP8). The aim of this study was to investigate whether 3,4,5-tri- O-caffeoylquinic acid (3,4,5-tri-CQA), isolated from propolis, shows a neuroprotective effect against Aβ-induced cell death on human neuroblastoma SH-SY5Y cells. To clarify the possible mechanism, we performed proteomics and real-time RT-PCR as well as a measurement of the intracellular adenosine triphosphate (ATP) level. These results showed that 3,4,5-tri-CQA attenuated the cytotoxicity and prevented Aβ-mediated apoptosis. Glycolytic enzymes, phosphoglycerate mutase 1 (PGAM1) and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) were overexpressed in co-treated cells with both 3,4,5-tri-CQA and Aβ. The mRNA expression of PGAM1, G3PDH, and phosphoglycerate kinase 1 (PGK1), and intracellular ATP level were also increased in 3,4,5-tri-CQA treated cells. Taken together the findings in our study suggests that 3,4,5-tri-CQA shows a neuroprotective effect against Aβ-induced cell death through the upregulation of glycolytic enzyme mRNA as well as ATP production activation. [ABSTRACT FROM AUTHOR]
- Published
- 2011
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61. p19 ras Represses proliferation of non-small cell lung cancer possibly through interaction with Neuron-Specific Enolase (NSE)
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Jang, Sang-Min, Kim, Jung-Woong, Kim, Chul-Hong, Kim, Daehwan, Rhee, Sangmyung, and Choi, Kyung-Hee
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SMALL cell lung cancer , *ONCOGENES , *ENOLASE , *MESSENGER RNA , *CANCER cell proliferation , *CARRIER proteins , *PROTEIN-protein interactions , *GLYCOLYSIS , *DIAGNOSIS - Abstract
Abstract: p19 ras is an alternative splicing product of the proto-oncogene c-H-ras pre-mRNA. In this study, we identified a novel p19 ras -binding protein, Neuron-Specific Enolase (NSE), using the yeast two-hybrid method. NSE is one of the enolase families that convert 2-phospho-d-glycerate (PGA) to phosphoenolpyruvate (PEP) in the glycolysis pathway. In both endogenous and over-expressed systems, we confirmed interactions between p19 ras and NSE via co-immunoprecipitation assay. We also identified the interaction region of p19 ras , which is required for binding with NSE. When full-length p19 ras and C-terminal region are bound to NSE, it inhibits the enzymatic activity of NSE. Furthermore, p19 ras interacted with Enolase α (Enoα) and repressed its enzymatic activity in vitro. p19 ras repressed lung cancer cell proliferation mostly increased by NSE in H1299 cells. Taken together, these results suggest that p19 ras is a novel regulator to suppress cell proliferation in lung cancer through the interaction with NSE. [Copyright &y& Elsevier]
- Published
- 2010
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62. Direct Binding of Glyceraldehyde 3-Phosphate Dehydrogenase to Telomeric DNA Protects Telomeres against Chemotherapy-Induced Rapid Degradation
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Demarse, Neil A., Ponnusamy, Suriyan, Spicer, Eleanor K., Apohan, Elif, Baatz, John E., Ogretmen, Besim, and Davies, Christopher
- Subjects
- *
PHOSPHATES , *DEHYDROGENASES , *DNA , *TELOMERES , *DRUG therapy , *DOXORUBICIN , *CHROMATIN - Abstract
Abstract: Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a glycolytic enzyme that displays several non-glycolytic activities, including the maintenance and/or protection of telomeres. In this study, we determined the molecular mechanism and biological role of the interaction between GAPDH and human telomeric DNA. Using gel-shift assays, we show that recombinant GAPDH binds directly with high affinity (K d =45 nM) to a single-stranded oligonucleotide comprising three telomeric DNA repeats, and that nucleotides T1, G5, and G6 of the TTAGGG repeat are essential for binding. The stoichiometry of the interaction is 2:1 (DNA:GAPDH), and GAPDH appears to form a high-molecular-weight complex when bound to the oligonucleotide. Mutation of Asp32 and Cys149, which are localized to the NAD-binding site and the active-site center of GAPDH, respectively, produced mutants that almost completely lost their telomere-binding functions both in vitro and in situ (in A549 human lung cancer cells). Treatment of A549 cells with the chemotherapeutic agents gemcitabine and doxorubicin resulted in increased nuclear localization of expressed wild-type GAPDH, where it protected telomeres against rapid degradation, concomitant with increased resistance to the growth-inhibitory effects of these drugs. The non-DNA-binding mutants of GAPDH also localized to the nucleus when expressed in A549 cells, but did not confer any significant protection of telomeres against chemotherapy-induced degradation or growth inhibition; this occurred without the involvement of caspase activation or apoptosis regulation. Overall, these data demonstrate that GAPDH binds telomeric DNA directly in vitro and may have a biological role in the protection of telomeres against rapid degradation in response to chemotherapeutic agents in A549 human lung cancer cells. [Copyright &y& Elsevier]
- Published
- 2009
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63. 3D structure of phosphofructokinase from Pichia pastoris: Localization of the novel γ-subunits
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Benjamin, Shaun, Radermacher, Michael, Kirchberger, Jürgen, Schöneberg, Torsten, Edelmann, Anke, and Ruiz, Teresa
- Subjects
- *
PICHIA pastoris , *PHOSPHOFRUCTOKINASE 1 , *ELECTRON microscopy , *GLYCOLYSIS , *METHYLOTROPHIC microorganisms , *TOMOGRAPHY , *PLANT enzymes , *IMAGE processing - Abstract
Abstract: The largest and one of the most complex ATP-dependent allosteric phosphofructokinase (Pfk) has been found in the methylotrophic yeast, Pichia pastoris. The enzyme is a hetero-oligomer (∼1MDa) composed of three distinct subunits (α, β and γ) with molecular masses of 109, 104 and 41kDa, respectively. While the α- and β-subunits show sequence similarities to other phosphofructokinase subunits, the γ-subunit does not show high homology to any known protein in the databases. We have determined the first quaternary structure of P. pastoris phosphofructokinase by 3D electron microscopy. Random conical techniques and tomography have been instrumental to ascertain the quality of the sample preparations for structural studies and to obtain a reliable 3D structure. The final reconstruction of P. pastoris Pfk resembles its yeast counterparts with four additional densities, assigned to four γ-subunits, bridging the N-terminal domains of the four pairs of α- and β-subunits. Our data has evidenced novel interactions between the γ- and the α-subunits comparable in intensity to the interactions, shown by cross-linking and limited proteolytic degradation experiments, between the γ- and β-subunits. The structural data provides clear insights into the allosteric fine-tuned regulation of the enzyme by ATP and AMP observed in this yeast species. [Copyright &y& Elsevier]
- Published
- 2009
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64. Structures of S. pombe phosphofructokinase in the F6P-bound and ATP-bound states
- Author
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Benjamin, Shaun, Radermacher, Michael, Bär, Jörg, Edelmann, Anke, and Ruiz, Teresa
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PHOSPHOFRUCTOKINASE 1 , *FRUCTOSE , *ENZYMES , *MICROSCOPY - Abstract
Abstract: Phosphofructokinase (Pfk1; EC 2.7.1.11) is the third enzyme of the glycolytic pathway catalyzing the formation of fructose-1,6-bisphosphate from fructose-6-phosphate (F6P) and ATP. Schizosaccharomyces pombe Pfk1 is a homo-octameric enzyme of 800kDa molecular weight, distinct from its yeast counterparts which are mostly hetero-octameric enzymes composed of two different subunits. Having an “open” conformation and a tendency to aggregate into higher oligomeric structures, the S. pombe enzyme shows similarities to the mammalian muscle Pfk1. It has been proposed that due to the distinct N-terminal region of the S. pombe subunit, the oligomeric organization of subunits in this enzyme is different from other yeast phosphofructokinases. Electron microscopy studies were carried out to reveal the quaternary structure of the homo-octameric Pfk1 from S. pombe in the F6P-bound and in the ATP-bound state. Random conical tilt data sets have been collected from deep stain preparations of the enzyme in both states. The 0° tilt images have been separated into different classes and a 3D reconstruction has been calculated for each class from the high tilt images. Our results confirm the presence of a variety of views of the particle, most of which can be interpreted as views of the molecule rotating around its long axis. Despite the biochemical differences, the structure of phosphofructokinase from S. pombe in the presence of either F6P or ATP is similar to the hetero-octameric structure of phosphofructokinase from Saccharomyces cerevisiae. The molecule can be described as composed of two subdomains, connected by two well-defined densities. We have been able to establish a correlation between the kinetic behavior and the structural conformation of Pfk1. [Copyright &y& Elsevier]
- Published
- 2007
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65. Survey of bacterial proteins released in cheese: a proteomic approach
- Author
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Gagnaire, Valérie, Piot, Michel, Camier, Bénédicte, Vissers, Johannes P.C., Jan, Gwénaël, and Léonil, Joëlle
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BACTERIAL proteins , *CHEESE , *TASTE testing of food , *PROTEOMICS - Abstract
During the ripening of Emmental cheese, the bacterial ecosystem confers its organoleptic characteristics to the evolving curd both by the action of the living cells, and through the release of numerous proteins, including various types of enzymes into the cheese when the cells lyse. In Emmental cheese these proteins can be released from thermophilic lactic acid bacteria used as starters like Lactobacillus helveticus, Lb delbruecki subsp. lactis and Streptococcus salivarius subsp. thermophilus and ripening bacteria such as Propionibacterium freudenreichii. The aim of this study was to obtain a proteomic view of the different groups of proteins within the cheese using proteomic tools to create a reference map. A methodology was therefore developed to reduce the complexity of cheese matrix prior to 2D-PAGE analysis. The aqueous phase of cheese was prefractionated by size exclusion chromatography, bacterial and milk proteins were separated and subsequently characterised by mass spectrometry, prior to peptide mass fingerprint and sequence homology database search. Five functional groups of proteins were identified involved in: (i) proteolysis, (ii) glycolysis, (iii) stress response, (iv) DNA and RNA repair and (v) oxidoreduction. The results revealed stress responses triggered by thermophilic lactic acid bacteria and Propionibacterium strains at the end of ripening.Information was also obtained regarding the origin and nature of the peptidases released into the cheese, thus providing a greater understanding of casein degradation mechanisms during ripening. Different peptidases arose from St thermophilus and Lb helveticus, suggesting that streptococci are involved in peptide degradation in addition to the proteolytic activity of lactobacilli. [Copyright &y& Elsevier]
- Published
- 2004
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66. Localization of enolase in synaptic plasma membrane as an αγ heterodimer in rat brain
- Author
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Ueta, Hisashi, Nagasawa, Hideko, Oyabu-Manabe, Yuriko, Toida, Kazunori, Ishimura, Kazunori, and Hori, Hitoshi
- Subjects
- *
CELL membranes , *BRAIN research , *SYNAPTOSOMES , *DIMERS - Abstract
Enolase, a glycolytic enzyme, is a multifunctional protein with location diversity. We revealed the intracellular distribution of enolase isozymes, such as αα-, αγ- and γγ-enolases, in rat brain synaptic terminals by biochemical and immunoelectron microscopic analyses. Specific activity of enolase of synaptic plasma membrane fraction (SPM2) obtained from synaptosomes was
23.2±4.4×10−2 μmol/mg protein/min in the presence of 0.25% Triton X-100 and that of synaptosomal cytoplasm (LS) was67.4±12.1×10−2 μmol/mg protein/min. About half of enolase activity in synaptosomes was distributed to soluble fraction while the remaining stayed in particulate membrane fractions by ultracentrifugation. Immunoblot analysis of the fractions demonstrated both α and γ subunits were distributed in SPM. In addition, immunoelectron microscopic analysis also revealed that both subunits were immunoreactive on the SPM. Using coimmunoprecipitation assay, we confirmed that the enolase was present not only as a homodimer form but also as an αγ hybrid form associated with membrane, where both subunits were coimmunoprecipitated from lysate of SPM2 in the presence of Mg2+. These findings indicate that all forms (αα, αγ, and γγ) of enolase translocate to the plasma membrane and associate with some components in the SPM. [Copyright &y& Elsevier]- Published
- 2004
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67. BcI-2 Inhibits Tumor Necrosis Factor-α-mediated Increase of Blycolytic Enzyme Activities and Enhances Pyruvate Carboxylase Activity.
- Author
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Yeon Hyang Kim and Soung Soo Kim
- Subjects
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GLUCOSE , *METABOLISM , *TUMOR necrosis factors , *CELL-mediated cytotoxicity , *ENZYMES , *CELLS , *LACTATE dehydrogenase - Abstract
To understand the effects of bcl-2 on glucose metabolism and tumor necrosis factor-α (TNF-α) mediated cytotoxicity, the activities of glycolytic enzymes (hexokinase, 6-phosphofructo-1-kinase, and pyruvate kinase), lactate dehydrogenase, pyruvate carboxylase, and phosphoenolpyruvate carboxykinase were examined with or without TNF-α treatment in TNF-α sensitive 1,929 cells and TNF-α resistant bcl-2 transfected L929 cells. In TNF-α-treated L929 cells, the activities of the glycolytic enzymes and lactate dehydrogenase greatly increased, but there was no detectable change in phosphoenolpyruvate carboxykinase. Pyruvate carboxylase activity decreased by about 25% between 6 and 12 h after TNF-α treatment. The activities of the glycol)tic enzymes and lactate dehydrogenase in bcl-2 transletted L929 cells were lower than in L929 cells upon TNF-α treatment. On the other hand, the activity of pyruvate carboxylase was 20-100% greater after 6 h of TNF-α treatment than in the L929 cells. The activity of phosphoenolpyruvate carboxykinase of bcl-2 trasfected L929 cells was louver by up to 25% than in L929 cells after 12 h. The increase of pyruvate carboxylase activity and decrease of phosphoenolpyruvate carboxykinase activity in bcl-2 transfected 1,929 cells may contribute to the protective effects of bcl-2 against TNF-α mediated cytotoxicity. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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68. The 10.8-A˚ structure of Saccharomyces cerevisiae phosphofructokinase determined by cryoelectron microscopy: localization of the putative fructose 6-phosphate binding sites
- Author
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Ruiz, Teresa, Mechin, Ingrid, Bär, Jörg, Rypniewski, Wojciech, Kopperschläger, Gerhard, and Radermacher, Michael
- Subjects
- *
PHOSPHOFRUCTOKINASE 1 , *PHOSPHORYLATION , *FRUCTOSE , *SACCHAROMYCES cerevisiae - Abstract
Phosphofructokinase plays a key role in the regulation of the glycolytic pathway and is responsible for the phosphorylation of fructose 6-phosphate to fructose 1,6-bisphosphate. Although the structure of the bacterial enzyme is well understood, the knowledge is still quite limited for higher organisms given the larger size and complexity of the eukaryotic enzymes. We have studied phosphofructokinase from Saccharomyces cerevisiae in the presence of fructose 6-phosphate by cryoelectron microscopy and image analysis of single particles and obtained the structure at 10.8 A˚ resolution. This was achieved by optimizing the illumination conditions to obtain routinely 8-A˚ data from hydrated samples in an electron microscope equipped with an LaB6 and by improving the image alignment techniques. The analysis of the structure has evidenced that the homology of the subunits at the sequence level has transcended to the structural level. By fitting the X-ray structure of the bacterial tetramer into each dimer of the yeast octamer the putative binding sites for fructose 6-phosphate were revealed. The data presented here in combination with molecular replacement techniques have served to provide the initial phases to solve the X-ray structure of the yeast phosphofructokinase. [Copyright &y& Elsevier]
- Published
- 2003
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69. Effect of herb supplement on hepatic enzyme activities in ddY mice.
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Arai, T., Horii, N., Koshirakawa, M., Nakajyo, S., and Sako, T.
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GLUCOSE ,BLOOD sugar ,LIPIDS ,TRIGLYCERIDES ,HERBAL medicine ,FATTY acids ,ENZYMES ,MICE - Abstract
Plasma glucose and lipid concentrations and hepatic enzyme activities were measured in male ddY mice supplemented with the herb, Echevaria glauca, to examine the effect of herbal treatment. In mice supplemented with the herb, plasma triglyceride (TG) and free fatty acid (FFA) concentrations decreased and hepatic glycolytic enzyme and glutathione peroxidase (GSHpx) activities increased significantly compared with those in the nontreated control mice. These increases in hepatic enzyme activities were not fully dosedependent, however the higher dose and longer duration with herb supplement induced increases in the enzyme activities. It was found that dietary herb supplement caused an acceleration of hepatic function, judged by increased activities of glycolytic enzyme and GSHpx in ddY mice. [ABSTRACT FROM AUTHOR]
- Published
- 2001
- Full Text
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70. Implication of Potential Differential Roles of the Two Phosphoglucomutase Isoforms in the Protozoan Parasite Cryptosporidium parvum .
- Author
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Nie J, Yin J, Wang D, Wang C, and Zhu G
- Abstract
Phosphoglucomutase 1 (PGM1) catalyzes the conversion between glucose-1-phosphate and glucose-6-phosphate in the glycolysis/glucogenesis pathway. PGM1s are typically cytosolic enzymes in organisms lacking chloroplasts. However, the protozoan Cryptosporidium parasites possess two tandemly duplicated PGM1 genes evolved by a gene duplication after their split from other apicomplexans. Moreover, the downstream PGM1 isoform contains an N-terminal signal peptide, predicting a non-cytosolic location. Here we expressed recombinant proteins of the two PGM1 isoforms from the zoonotic Cryptosporidium parvum , namely CpPGM1A and CpPGM1B, and confirmed their enzyme activity. Both isoforms followed Michaelis-Menten kinetics towards glucose-1-phosphate ( K
m = 0.17 and 0.13 mM, Vmax = 7.30 and 2.76 μmol/min/mg, respectively). CpPGM1A and CpPGM1B genes were expressed in oocysts, sporozoites and intracellular parasites at a similar pattern of expression, however CpPGM1A was expressed at much higher levels than CpPGM1B . Immunofluorescence assay showed that CpPGM1A was present in the cytosol of sporozoites, however this was enriched towards the plasma membranes in the intracellular parasites; whereas CpPGM1B was mainly present under sporozoite pellicle, although relocated to the parasitophorous vacuole membrane in the intracellular development. These observations indicated that CpPGM1A played a house-keeping function, while CpPGM1B played a different biological role that remains to be defined by future investigations.- Published
- 2021
- Full Text
- View/download PDF
71. Cotransport of Glyceraldehyde-3-Phosphate Dehydrogenase and Actin in Axons of Chicken Motoneurons.
- Author
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Yuan, Aidong, Mills, Roland, Bamburg, James, and Bray, John
- Abstract
1.To study proteins transported with actin in axons, we pulse-labeled motoneurons in the chicken sciatic nerve with [
35 S]methionine and, 1–20 days later, isolated actin and its binding proteins by affinity chromatography of Triton soluble nerve extracts on DNase I–Sepharose. The DNase I-purified proteins were electrophoresed on two-dimensional gels and the specific activity of the radioactively labeled protein spots was estimated by fluorography. 2.In addition to actin, which binds specifically to DNase I, a small number of other proteins were labeled, including established actin monomer binding proteins and a protein of 36 kDa and p I 8.5. On the basis of its molecular mass, p I, amino acid composition, and immunostaining, the unrecognized protein was identified as the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). 3.The high-affinity binding of GAPDH to actin was confirmed by incubation of Triton-soluble nerve extracts with either mouse anti-GAPDH (or antiactin) and indirect immunomagnetic separation with Dynabeads covalently linked to sheep anti-mouse antibody. Analysis by one-dimensional gel electrophoresis and immunoblotting showed that actin and GAPDH were the main proteins isolated by these methods. 4.Analysis of labeled nerves at 12 and 20 days after pulse labeling showed that GAPDH and actin were transported at the same rate, i.e., 3–5 mm/day, which corresponds to slow component b of axonal transport. These proteins were not associated with rapidly transported proteins that accumulated proximal to a ligation 7 cm from the spinal cord 9 hr after injection of radioactivity. 5.Our results indicate that GAPDH and actin are transported as a complex in axons and raise the possibility that GAPDH could act as a chaperone for monomeric actin, translocating it to intraaxonal sites for exchange with or assembly into actin filaments. Alternatively, actin could be involved in translocating and anchoring GAPDH to specialized sites in axons and nerve terminals that require a source of ATP by glycolysis. [ABSTRACT FROM AUTHOR]- Published
- 1999
- Full Text
- View/download PDF
72. Ambiquitous behavior of rabbit liver lactate dehydrogenase.
- Author
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Sanz, M. and Lluis, C.
- Abstract
Rabbit liver mitochondrial fraction shows lactate dehydrogenase activity. The enzyme can be released from particles by increasing the pH and the ionic strength of the medium. There is a narrow range of pH (6.8-7.4) and ionic strength (20-50 mM NaCl) in which the solubilization sharply increases. It has been shown that divalent anions (SO) and cations (Mg, Ca) are highly effective specific solubilizing agents. NADH (1.5 mM) and ATP (1.0 mM) were effective in solubilizing 50% of the enzyme bound, whereas the same concentrations of the analogs NAD and ADP had little effect. Cytosolic lactate dehydrogenase bound to the mitochondrial fraction and a saturation of particles by enzyme was observed in all experiments performed. The in vitro binding requires a short period of incubation between the enzyme and particles and the binding is independent of the temperature in the 0-37°C range. Binding was prevented by 0.15 M NaCl. The bound enzyme is approximately 20% less active than the soluble one. The results described give support to the proposal that rabbit liver lactate dehydrogenase has an ambiquitous behavior, like other glycolytic enzymes, which have not a fixed intracellular localization. [ABSTRACT FROM AUTHOR]
- Published
- 1988
- Full Text
- View/download PDF
73. Sperm-Specific Glycolysis Enzyme Glyceraldehyde-3-Phosphate Dehydrogenase Regulated by Transcription Factor SOX10 to Promote Uveal Melanoma Tumorigenesis.
- Author
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Ding X, Wang L, Chen M, Wu Y, Ge S, Li J, Fan X, and Lin M
- Abstract
Melanoma cells exhibit increased aerobic glycolysis, which represents a major biochemical alteration associated with malignant transformation; thus, glycolytic enzymes could be exploited to selectively target cancer cells in cancer therapy. Sperm-specific glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) switches glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate by coupling with the reduction of NAD+ to NADH. Here, we demonstrated that GAPDHS displays significantly higher expression in uveal melanoma (UM) than in normal controls. Functionally, the knockdown of GAPDHS in UM cell lines hindered glycolysis by decreasing glucose uptake, lactate production, adenosine triphosphate (ATP) generation, cell growth and proliferation; conversely, overexpression of GAPDHS promoted glycolysis, cell growth and proliferation. Furthermore, we identified that SOX10 knockdown reduced the activation of GAPDHS, leading to an attenuated malignant phenotype, and that SOX10 overexpression promoted the activation of GAPDHS, leading to an enhanced malignant phenotype. Mechanistically, SOX10 exerted its function by binding to the promoter of GAPDHS to regulate its expression. Importantly, SOX10 abrogation suppressed in vivo tumor growth and proliferation. Collectively, the results reveal that GAPDHS, which is regulated by SOX10, controls glycolysis and contributes to UM tumorigenesis, highlighting its potential as a therapeutic target., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ding, Wang, Chen, Wu, Ge, Li, Fan and Lin.)
- Published
- 2021
- Full Text
- View/download PDF
74. Comparison of activity, expression and S-nitrosylation of glycolytic enzymes between pale, soft and exudative and red, firm and non-exudative pork during post-mortem aging.
- Author
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Wang, Yingying, Liu, Rui, Hou, Qin, Tian, Xiaona, Fan, Xiaoquan, Zhang, Wangang, and Zhou, Guanghong
- Subjects
- *
NITRIC-oxide synthases , *GLYCOGEN phosphorylase , *CALPAIN , *PORK , *PYRUVATE kinase , *ENZYMES , *LACTIC acid - Abstract
• The method has significant advantage in identifying S-nitrosylated protein. • S-nitrosylation of those enzymes for postmortem meat is still unknown. • Protein S-nitrosylation may exert effect on glycolysis. • It is a new insight to explain PSE meat in postmortem muscle. The activity, expression and S-nitrosylation of glycogen phosphorylase (GP), phosphofructokinase (PFK) and pyruvate kinase (PK) was compared between pale, soft and exudative (PSE) and red, firm and non-exudative (RFN) pork. The nitric oxide synthase (NOS) activity of RFN pork was higher than PSE pork (P < 0.05). Glycogen and lactic acid content were significantly different between PSE and RFN samples at 1 h postmortem (P < 0.05). Compared to PSE pork, RFN pork had lower activities and higher S-nitrosylation levels of GP, PFK and PK (P < 0.05). Moreover, GP expression in RFN pork was lower (P < 0.05) while no significant differences of PFK and PK expression were observed between these two groups. These data suggest that protein S-nitrosylation can presumably regulate glycolysis by modulating glycolytic enzymes activities and then regulate the development of PSE pork. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
75. Triose phosphate isomerase from the blood flukeSchistosoma mansoni: Biochemical characterisation of a potential drug and vaccine target
- Author
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David J. Timson, David W. Dunne, Edward J. Farnell, and Veronika L Zinsser
- Subjects
Models, Molecular ,Vaccine target ,Proteolysis ,030231 tropical medicine ,Biophysics ,Glycolytic enzyme ,Blood fluke ,Bilharzia ,Glyceraldehyde 3-Phosphate ,Biochemistry ,Triosephosphate isomerase ,Epitopes ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Structural Biology ,Glyceraldehyde ,Genetics ,medicine ,Animals ,Humans ,Schistosomiasis ,Glycolysis ,Amino Acid Sequence ,Enzyme kinetics ,Molecular Biology ,030304 developmental biology ,Dihydroxyacetone phosphate ,chemistry.chemical_classification ,0303 health sciences ,medicine.diagnostic_test ,biology ,Schistosoma mansoni ,Cell Biology ,biology.organism_classification ,Molecular biology ,3. Good health ,Kinetics ,Enzyme ,chemistry ,Dihydroxyacetone Phosphate ,Antigens, Helminth ,Triose-Phosphate Isomerase - Abstract
The glycolytic enzyme triose phosphate isomerase from Schistosoma mansoni is a potential target for drugs and vaccines. Molecular modelling of the enzyme predicted that a Ser-Ala-Asp motif which is believed to be a helminth-specific epitope is exposed. The enzyme is dimeric (as judged by gel filtration and cross-linking), resistant to proteolysis and highly stable to thermal denaturation (melting temperature of 82.0 °C). The steady-state kinetic parameters are high (Km for dihydroxyacetone phosphate is 0.51 mM; Km for glyceraldehyde 3-phosphate is 1.1 mM; kcat for dihydroxyacetone phosphate is 7800 s(-1) and kcat for glyceraldehyde 3-phosphate is 6.9s(-1)).
- Published
- 2013
76. Cytoplasmic Organisation and the Properties of Cell Water: Speculations on Animal Cell Cryopreservation
- Author
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Clegg, J. S., Pegg, David E., editor, and Karow, Armand M., Jr., editor
- Published
- 1987
- Full Text
- View/download PDF
77. Implications of Metabolic Compartmentation in Prokaryotic Cells
- Author
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Moses, V., Welch, G. Rickey, editor, and Clegg, James S., editor
- Published
- 1986
- Full Text
- View/download PDF
78. The Cytoskeleton
- Author
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Osborn, Mary, Weber, Klaus, Welch, G. Rickey, editor, and Clegg, James S., editor
- Published
- 1986
- Full Text
- View/download PDF
79. Biochemical Mechanisms of Pentostam
- Author
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Berman, Jonathan D., Grogl, Max, and Hart, D. T., editor
- Published
- 1989
- Full Text
- View/download PDF
80. A Multicomponent, Random Walk Model of Transport and Metabolism Inside a Neuron
- Author
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Kufahl, R. H., Hanley, T. R., Bruley, D. F., Halsey, J. H., Jr., and Longmuir, Ian S., editor
- Published
- 1986
- Full Text
- View/download PDF
81. Stimulation of the Synthesis of Fructose 1,6-Diphosphate Aldolase by Transferrin
- Author
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Oh, T. H., Markelonis, G. J., Guidera, T. Dion, Hobbs, S. L., Park, L. P., Strohman, Richard C., editor, and Wolf, Stewart, editor
- Published
- 1985
- Full Text
- View/download PDF
82. The Evolutionary Origin of Glycosomes: How Glycolysis Moved from Cytosol to Organelle in Evolution
- Author
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Borst, P., Swinkels, B. W., Grunberg-Manago, Marianne, editor, Clark, Brian F. C., editor, and Zachau, Hans G., editor
- Published
- 1989
- Full Text
- View/download PDF
83. Structure and Expression of Yeast Glycolytic Genes
- Author
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Holland, Michael J., Holland, Janice P., McAllister, Lee, Hollaender, Alexander, editor, DeMoss, Ralph D., editor, Kaplan, Samuel, editor, Konisky, Jordan, editor, Savage, Dwayne, editor, and Wolfe, Ralph S., editor
- Published
- 1982
- Full Text
- View/download PDF
84. End-Product Tolerance and Ethanol
- Author
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Rose, Anthony H., Beavan, Michael J., Hollaender, Alexander, editor, Rabson, Robert, editor, Rogers, Palmer, editor, Pietro, Anthony San, editor, Valentine, Raymond, editor, and Wolfe, Ralph, editor
- Published
- 1981
- Full Text
- View/download PDF
85. The Cytoskeleton and the Cytoplasmic Matrix
- Author
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Schliwa, Manfred, Alfert, M., Beermann, W., Goldstein, L., Porter, K. R., and Schliwa, Manfred
- Published
- 1986
- Full Text
- View/download PDF
86. Biogenesis of Glycosomes (Microbodies) in the Trypanosomatidae
- Author
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Opperdoes, F. R., Fahimi, H. Dariush, editor, and Sies, Helmut, editor
- Published
- 1987
- Full Text
- View/download PDF
87. The significance of phosphofructokinase to the regulation of carbohydrate metabolism
- Author
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HOFMANN, E., Adrian, R. H., editor, Helmreich, E., editor, Holzer, H., editor, Jung, R., editor, Kramer, K., editor, Krayer, O., editor, Linden, R. J., editor, Lynen, F., editor, Miescher, P. A., editor, Piiper, J., editor, Rasmussen, H., editor, Renold, A. E., editor, Trendelenburg, U., editor, Ullrich, K., editor, Vogt, W., editor, and Weber, A., editor
- Published
- 1976
- Full Text
- View/download PDF
88. Regulation of Glycolysis
- Author
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Hess, B., Boiteux, A., Kuschmitz, D., Sund, Horst, editor, and Ullrich, Volker, editor
- Published
- 1983
- Full Text
- View/download PDF
89. Metabolic Changes in Long-Term Stimulated Fast Muscles
- Author
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Brown, M., Cotter, M., Hudlická, O., Smith, M., Vrbová, G., Howald, H., editor, and Poortmans, Jacques R., editor
- Published
- 1975
- Full Text
- View/download PDF
90. Integration of Mitochondrial Function with High Aerobic Glycolysis in Tumors: Role of Hexokinase Binding to the Outer Mitochondrial Membrane
- Author
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Nakashima, Richard A., Paggi, Marco G., Arora, Krishan K., Pedersen, Peter L., Lemasters, John J., editor, Hackenbrock, Charles R., editor, Thurman, Ronald G., editor, and Westerhoff, Hans V., editor
- Published
- 1988
- Full Text
- View/download PDF
91. The 'Cytosol': A Neglected and Poorly Understood Compartment of Eukaryotic Cells
- Author
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Clegg, J. S., Barrios, M. B., Cañedo, L. E., editor, Todd, L. E., editor, Packer, L., editor, and Jaz, J., editor
- Published
- 1988
- Full Text
- View/download PDF
92. Toward The Development Of New Drugs For Parasitic Diseases
- Author
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Opperdoes, Fred R., Warren, Kenneth S., editor, and Bowers, Jhon Z., editor
- Published
- 1983
- Full Text
- View/download PDF
93. Role of Multienzyme Complexes in the Integration of Cellular Metabolism
- Author
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Kurganov, B. I., Chock, P. Boon, editor, Huang, Charles Y., editor, Tsou, C. L., editor, and Wang, Jerry H., editor
- Published
- 1988
- Full Text
- View/download PDF
94. Biochemistry of Muscle Diseases
- Author
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Pennington, R. J. T. and Cumings, J. N., editor
- Published
- 1972
- Full Text
- View/download PDF
95. Identification of Alternative Allosteric Sites in Glycolytic Enzymes for Potential Use as Species-Specific Drug Targets.
- Author
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Ayyildiz M, Celiker S, Ozhelvaci F, and Akten ED
- Abstract
Three allosteric glycolytic enzymes, phosphofructokinase, glyceraldehyde-3 phosphate dehydrogenase and pyruvate kinase, associated with bacterial, parasitic and human species, were explored to identify potential allosteric sites that would be used as prime targets for species-specific drug design purposes using a newly developed approach which incorporates solvent mapping, elastic network modeling, sequence and structural alignments. The majority of binding sites detected by solvent mapping overlapped with the interface regions connecting the subunits, thus appeared as promising target sites for allosteric regulation. Each binding site was then evaluated by its ability to alter the global dynamics of the receptor defined by the percentage change in the frequencies of the lowest-frequency modes most significantly and as anticipated, the most effective ones were detected in the vicinity of the well-reported catalytic and allosteric sites. Furthermore, some of our proposed regions intersected with experimentally resolved sites which are known to be critical for activity regulation, which further validated our approach. Despite the high degree of structural conservation encountered between bacterial/parasitic and human glycolytic enzymes, the majority of the newly presented allosteric sites exhibited a low degree of sequence conservation which further increased their likelihood to be used as species-specific target regions for drug design studies., (Copyright © 2020 Ayyildiz, Celiker, Ozhelvaci and Akten.)
- Published
- 2020
- Full Text
- View/download PDF
96. Proteome Changes during Meat Aging in Tough and Tender Beef Suggest the Importance of Apoptosis and Protein Solubility for Beef Aging and Tenderization
- Author
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Christophe Chambon, Jean-François Hocquette, Elisabeth Laville, Martine Morzel, Jacques Lepetit, Sylvie Blinet, Gilles Renand, Thierry Sayd, Qualité des Produits Animaux (QuaPA), Institut National de la Recherche Agronomique (INRA), FLAveur, VIsion et Comportement du consommateur (FLAVIC), Institut National de la Recherche Agronomique (INRA)-Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD)-Université de Bourgogne (UB), Station de Génétique Quantitative et Appliquée (SGQA), Unité de Recherches sur les Herbivores (URH), and This study was funded by a national grant from the Agence Nationale de la Recherche and from APIS-GENE (GENANIMAL call, national program AGENAE) for the project MUGENE (GENEs of the MUscle tissue).
- Subjects
Male ,Time Factors ,hierarchical clustering analysis ,Muscle Proteins ,Apoptosis ,chemistry.chemical_compound ,[SDV.IDA]Life Sciences [q-bio]/Food engineering ,Electrophoresis, Gel, Two-Dimensional ,Food science ,Solubility ,chemistry.chemical_classification ,0303 health sciences ,education.field_of_study ,Food preservation ,protein solubility ,04 agricultural and veterinary sciences ,glycolytic enzyme ,proteome analysis ,Mitochondria ,Tenderness ,tenderness ,Proteome ,medicine.symptom ,Shear Strength ,hsp27 ,General Agricultural and Biological Sciences ,Tris ,proteolysis ,Meat ,Population ,Biology ,03 medical and health sciences ,mitochondrial membrane ,medicine ,Animals ,Muscle, Skeletal ,education ,030304 developmental biology ,Chromatography ,beef muscle ,0402 animal and dairy science ,General Chemistry ,040201 dairy & animal science ,Enzyme ,chemistry ,Postmortem Changes ,Food Technology ,Cattle - Abstract
Chantier qualité GA; International audience; Within a population of Charolais young bulls, two extreme groups of longissimus thoracis muscle samples, classified according to Warner−Bratzler shear force (WBSF) of 55 °C grilled meat, were analyzed by 2D-electrophoresis. Muscle analyses were performed on 4 bulls of the “tender” group (WBSF = 27.7 ± 4.8 N) and 4 bulls of the “tough” group (WBSF = 41.2 ± 6.1 N), at 3 post-mortem times: D0, samples taken within 10 min post-mortem; D5 and D21, samples kept at 4 °C under vacuum during 5 and 21 days. Proteins of muscle samples were separated in two fractions based on protein solubility in Tris buffer: “soluble” and “insoluble”. Proteins of both fractions were separated by 2D-electrophoresis. Evolution of spots during the 3 post-mortem times was analyzed by hierarchical classification (HCA). Three clusters of proteins presenting similar evolution profiles provided accurate classification of post-mortem times and showed the translocation of some chaperone proteins and glycolytic enzymes from the soluble fraction to the insoluble fraction between D0 and D5. Cellular structure dismantlement and proteolysis was observed at D21. Effect of group (“tender” vs “tough”) on spot intensities was tested by ANOVA. At D0, higher quantity of proteins of the inner and outer membrane of mitochondria was found in the tender group suggesting a more extensive degradation of mitochondria that may be related to the apoptotic process.
- Published
- 2009
97. Molecular and biochemical characterisation of Trypanosoma cruzi phosphofructokinase
- Author
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Noelia Lander, Evelyn Rodriguez, and Jose Luis Ramirez
- Subjects
Microbiology (medical) ,lcsh:Arctic medicine. Tropical medicine ,lcsh:RC955-962 ,Trypanosoma cruzi ,lcsh:QR1-502 ,Biology ,Trypanosoma brucei ,lcsh:Microbiology ,Leishmania mexicana ,Structure-Activity Relationship ,parasitic diseases ,molecular biochemical characterisation ,Cloning, Molecular ,Kinetoplastida ,Gene ,Peptide sequence ,Molecular mass ,DNA, Protozoan ,glycolytic enzyme ,biology.organism_classification ,Molecular biology ,Open reading frame ,Microscopy, Fluorescence ,Phosphofructokinases ,phosphofructokinase ,Biochemistry ,Phosphofructokinase - Abstract
The characterisation of the gene encoding Trypanosoma cruzi CL Brener phosphofructokinase (PFK) and the biochemical properties of the expressed enzyme are reported here. In contradiction with previous reports, the PFK genes of CL Brener and YBM strain T. cruzi were found to be similar to their Leishmania mexicana and Trypanosoma brucei homologs in terms of both kinetic properties and size, with open reading frames encoding polypeptides with a deduced molecular mass of 53,483. The predicted amino acid sequence contains the C-terminal glycosome-targeting tripeptide SKL; this localisation was confirmed by immunofluorescence assays. In sequence comparisons with the genes of other eukaryotes, it was found that, despite being an adenosine triphosphate-dependent enzyme, T. cruzi PFK shows significant sequence similarity with inorganic pyrophosphate-dependent PFKs.
- Published
- 2009
98. GAPDH Overexpression in the T Cell Lineage Promotes Angioimmunoblastic T Cell Lymphoma through an NF-κB-Dependent Mechanism.
- Author
-
Mondragón, Laura, Mhaidly, Rana, De Donatis, Gian Marco, Tosolini, Marie, Dao, Pascal, Martin, Anthony R., Pons, Caroline, Chiche, Johanna, Jacquin, Marie, Imbert, Véronique, Proïcs, Emma, Boyer, Laurent, Doye, Anne, Luciano, Frédéric, Neels, Jaap G., Coutant, Frédéric, Fabien, Nicole, Sormani, Laura, Rubio-Patiño, Camila, and Bossowski, Jozef P.
- Subjects
- *
T cells , *TRANSGENIC mice , *T helper cells , *LYMPHOMAS , *HUMAN T cells - Abstract
GAPDH is emerging as a key player in T cell development and function. To investigate the role of GAPDH in T cells, we generated a transgenic mouse model overexpressing GAPDH in the T cell lineage. Aged mice developed a peripheral Tfh-like lymphoma that recapitulated key molecular, pathological, and immunophenotypic features of human angioimmunoblastic T cell lymphoma (AITL). GAPDH induced non-canonical NF-κB pathway activation in mouse T cells, which was strongly activated in human AITL. We developed a NIK inhibitor to reveal that targeting the NF-κB pathway prolonged AITL-bearing mouse survival alone and in combination with anti-PD-1. These findings suggest the therapeutic potential of targeting NF-κB signaling in AITL and provide a model for future AITL therapeutic investigations. • Overexpression of GAPDH in T cell lineage in a mouse model recapitulates AITL disease • AITL tumors are characterized by a GAPDH induction of the NF-κB pathway • plck-GAPDH mice allow to model AITL disease for testing of new therapeutic strategies Mondragón et al. find that transgenic mice overexpressing GAPDH in T cells develop lymphoma that recapitulates key features of human angioimmunoblastic T cell lymphoma (AITL), including activation of the non-canonical NF-κB pathway. Blocking NF-κB activity with an NIK inhibitor reduces human and mouse AITL growth. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
99. Application of proteomics to the characterisation of milk and dairy products
- Author
-
Joëlle Léonil, Valérie Gagnaire, María A. Manso, Gwénaël Jan, Science et Technologie du Lait et de l'Oeuf (STLO), Institut National de la Recherche Agronomique (INRA)-AGROCAMPUS OUEST, and Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)-Institut national d'enseignement supérieur pour l'agriculture, l'alimentation et l'environnement (Institut Agro)
- Subjects
[SDV.SA]Life Sciences [q-bio]/Agricultural sciences ,proteolysis ,Microorganism ,PROTEOMIC ,Dairy industry ,Bacterial genome size ,Proteomics ,proteolyse ,01 natural sciences ,Applied Microbiology and Biotechnology ,produit laitier ,Protein expression ,stress ,03 medical and health sciences ,MILK ,protéome ,030304 developmental biology ,PROTEOLYTIC ENZYME ,2. Zero hunger ,0303 health sciences ,CHEESE ,biology ,010401 analytical chemistry ,Proteolytic enzymes ,biology.organism_classification ,0104 chemical sciences ,enzyme ,dairy product ,Biochemistry ,GLYCOLYTIC ENZYME ,Proteome ,Bacteria ,Food Science - Abstract
The use of proteomic tools allows a global and dynamic view of proteins that are expressed by bacteria. As an increasing number of bacterial genomes is currently available for homology searches, it is now possible to use such techniques to screen proteins expressed by microorganisms used in various fermented foods. Proteomic tools are also useful to investigate protein heterogeneity in protein-rich foods. In this paper the use of proteomic tools for the characterisation of milk proteins and in the study of protein expression of lactic acid bacteria used for manufacture of dairy products is reviewed; particular attention is given applied to advances in proteomic techniques available. The particular case of proteomics applied to cheese as an example of a complex food matrix [a mixture of animal (milk) and microbial proteins] is discussed, focusing on a novel strategy that allows the study of the enzymatic machinery found in situ in cheese.
- Published
- 2005
100. A monoclonal antibody that inhibits Trypanosoma cruzi growth in vitro and its reaction with intracellular triosephosphate isomerase
- Author
-
Cortés-Figueroa, A.A., Pérez-Torres, A., Salaiza, N., Cabrera, N., Escalona-Montaño, A., Rondán, A., Aguirre-García, M., Gómez-Puyou, A., Pérez-Montfort, R., and Becker, I.
- Published
- 2008
- Full Text
- View/download PDF
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