-
The Journal of Biological Chemistry Feb 2008Isoenzymes of phosphoglycerate kinase in Trypanosoma brucei are differentially expressed in its two main life stages. This study addresses how the organism manages to...
Isoenzymes of phosphoglycerate kinase in Trypanosoma brucei are differentially expressed in its two main life stages. This study addresses how the organism manages to make sufficient amounts of the isoenzyme with the correct localization, which processes (transcription, splicing, and RNA degradation) control the levels of mRNAs, and how the organism regulates the switch in isoform expression. For this, we combined new quantitative measurements of phosphoglycerate kinase mRNA abundance, RNA precursor stability, trans splicing, and ribosome loading with published data and made a kinetic computer model. For the analysis of regulation we extended regulation analysis. Although phosphoglycerate kinase mRNAs are present at surprisingly low concentrations (e.g. 12 molecules per cell), its protein is highly abundant. Substantial control of mRNA and protein levels was exerted by both mRNA synthesis and degradation, whereas splicing and precursor degradation had little control on mRNA and protein concentrations. Yet regulation of mRNA levels does not occur by transcription, but by adjusting mRNA degradation. The contribution of splicing to regulation is negligible, as for all cases where splicing is faster than RNA precursor degradation.
Topics: Animals; Computer Simulation; Gene Expression Regulation; Genes, Protozoan; Isoenzymes; Kinetics; Models, Biological; Phosphoglycerate Kinase; Protozoan Proteins; RNA Splicing; RNA, Messenger; RNA, Protozoan; Ribosomes; Transcription, Genetic; Trypanosoma brucei brucei
PubMed: 17991737
DOI: 10.1074/jbc.M705782200 -
Biomolecular NMR Assignments Oct 2017Human phosphoglycerate kinase (PGK) is an energy generating glycolytic enzyme that catalyses the transfer of a phosphoryl group from 1,3-bisphosphoglycerate (BPG) to ADP...
Human phosphoglycerate kinase (PGK) is an energy generating glycolytic enzyme that catalyses the transfer of a phosphoryl group from 1,3-bisphosphoglycerate (BPG) to ADP producing 3-phosphoglycerate (3PG) and ATP. PGK is composed of two α/β Rossmann-fold domains linked by a central α-helix and the active site is located in the cleft formed between the N-domain which binds BPG or 3PG, and the C-domain which binds the nucleotides ADP or ATP. Domain closure is required to bring the two substrates into close proximity for phosphoryl transfer to occur, however previous structural studies involving a range of native substrates and substrate analogues only yielded open or partly closed PGK complexes. X-ray crystallography using magnesium trifluoride (MgF) as a isoelectronic and near-isosteric mimic of the transferring phosphoryl group (PO), together with 3PG and ADP has been successful in trapping human PGK in a fully closed transition state analogue (TSA) complex. In this work we report the H, N and C backbone resonance assignments of human PGK in the solution conformation of the fully closed PGK:3PG:MgF:ADP TSA complex. Assignments were obtained by heteronuclear multidimensional NMR spectroscopy. In total, 97% of all backbone resonances were assigned in the complex, with 385 out of a possible 399 residues assigned in the H-N TROSY spectrum. Prediction of solution secondary structure from a chemical shift analysis using the TALOS-N webserver is in good agreement with the published X-ray crystal structure of this complex.
Topics: Adenosine Diphosphate; Fluorides; Glyceraldehyde 3-Phosphate; Humans; Magnesium Compounds; Models, Molecular; Nuclear Magnetic Resonance, Biomolecular; Phosphoglycerate Kinase; Protein Binding; Protein Conformation, alpha-Helical
PubMed: 28866776
DOI: 10.1007/s12104-017-9758-3 -
Biomedicine & Pharmacotherapy =... May 2023Inter-α-trypsin inhibitor heavy chain 4 (ITI-H4) is one of the acute phase proteins and is mainly related with inflammatory diseases such as bacterial bloodstream...
Inter-α-trypsin inhibitor heavy chain 4 (ITI-H4) is one of the acute phase proteins and is mainly related with inflammatory diseases such as bacterial bloodstream infection and recurrent pregnancy loss (RPL). In a previous study, ITI-H4 was reported to be cleaved by kallikrein B1 (KLKB1) and its cleaved form induces the imbalance between pro- and anti-inflammatory cytokines. Therefore, in this study, putative substrates of ITI-H4 were isolated by immunoprecipitation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS) analysis. Of those, phosphoglycerate kinase 1 (PGK1) was found to be a binding protein of ITI-H4. PGK1 increases the level of ITI-H4 expression and blocks the cleavage of ITI-H4 mediated by KLKB1. It also inhibits pro-inflammatory response by inhibiting the JAK2/STAT3 signaling pathway. Therefore, PGK1, a novel binding partner of ITI-H4, is expected to have cellular functions in the pathogenesis of ITI-H4-related inflammatory diseases.
Topics: Pregnancy; Female; Humans; Cytokines; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Phosphoglycerate Kinase
PubMed: 36841032
DOI: 10.1016/j.biopha.2023.114437 -
European Journal of Biochemistry Apr 19781. The testis-specific isoenzyme of phosphoglycerate kinase (phosphoglycerate kinase B) has been isolated from ram testes using a procedure which separates it from... (Comparative Study)
Comparative Study
1. The testis-specific isoenzyme of phosphoglycerate kinase (phosphoglycerate kinase B) has been isolated from ram testes using a procedure which separates it from 'normal' phosphoglycerate kinase which is also present in testis tissue. The purification procedure is described. 2. The best preparations had no detectable impurity on electrophoresis, and had specific activities comparable with the same enzyme from other sources. 3. Kinetic studies indicated that the two isoenzymes have identical properties, within experimental error, for substrate affinity (for MgATP, 3-phosphoglycerate and MgADP), energy of activation and thermal denaturation. 4. The molecular weights of both isoenzymes were not distinguishably different from those previously reported, as measured by polyacrylamide/dodecylsulphate electrophoresis. The amino acid compositions showed only slight differences, and tryptic peptide maps showed that there was close homology of sequence. Starch gel electrophoresis at pH 6.5 indicates that the B isoenzyme has 1--2 more positive charges than the A. 5. Phosphoglycerate kinase A isolated from sheep muscle was shown, within experimental error, to be identical to the phosphoglycerate kinase A isolated from testis. 6. The results further substantiate the suggestion that the B isoenzyme is coded by a gene which was duplicated from the phosphoglycerate kinase A gene.
Topics: Amino Acids; Animals; Isoenzymes; Kinetics; Male; Molecular Weight; Phosphoglycerate Kinase; Sheep; Testis
PubMed: 639826
DOI: 10.1111/j.1432-1033.1978.tb12215.x -
Cell Cycle (Georgetown, Tex.) Jul 2016
Topics: Phosphoglycerate Kinase
PubMed: 27105392
DOI: 10.1080/15384101.2016.1179037 -
The Journal of Biological Chemistry Aug 2012Phosphoglycerate kinase (PGK), present on the surface of group B streptococcus (GBS), has previously been demonstrated to bind the host proteins actin and plasminogen....
Phosphoglycerate kinase (PGK), present on the surface of group B streptococcus (GBS), has previously been demonstrated to bind the host proteins actin and plasminogen. The actin and plasminogen binding sites of GBS-PGK were identified using truncated GBS-PGK molecules, followed by peptide mapping. These experiments identified two actin and plasminogen binding sites located between amino acids 126-134 and 204-208 of the 398-amino acid-long GBS-PGK molecule. Substitution of the lysine residues within these regions with alanine resulted in significantly reduced binding to both actin and plasminogen. In addition, conversion of the glutamic acid residue at amino acid 133 to proline, the amino acid found at this position for the PGK protein of Streptococcus pneumoniae, also resulted in significantly reduced binding to actin and plasminogen. These results demonstrate that the lysine residues at amino acid positions 126, 127, 130, 204, and 208 along with the glutamic acid residue at amino acid position 133 are necessary for actin and plasminogen binding by GBS-PGK.
Topics: Actins; Bacterial Proteins; Humans; Peptide Mapping; Phosphoglycerate Kinase; Plasminogen; Protein Binding; Streptococcus agalactiae; Streptococcus pneumoniae
PubMed: 22761440
DOI: 10.1074/jbc.M112.361261 -
Aging Jul 2020The neurotoxin MPP (1-methyl-4-phenylpyridinium ion) disrupts mitochondrial function leading to oxidative stress and neuronal death. Here we examine whether activation...
The neurotoxin MPP (1-methyl-4-phenylpyridinium ion) disrupts mitochondrial function leading to oxidative stress and neuronal death. Here we examine whether activation of the Keap1-Nrf2 cascade can protect SH-SY5Y neuroblastoma cells from MPP-induced cytotoxicity. Treatment of SH-SY5Y cells with CBR-470-1, an inhibitor of the glycolytic enzyme phosphoglycerate kinase 1 (PGK1), leads to methylglyoxal modification of Keap1, Keap1-Nrf2 disassociation, and increased expression of Nrf2 responsive genes. Pretreatment with CBR-470-1 potently attenuated MPP-induced oxidative injury and SH-SY5Y cell apoptosis. CBR-470-1 neuroprotection is dependent upon Nrf2, as Nrf2 shRNA or CRISPR/Cas9-mediated Nrf2 knockout, abolished CBR-470-1-induced SH-SY5Y cytoprotection against MPP. Consistent with these findings, PGK1 depletion or knockout mimicked CBR-470-1-induced actions and rendered SH-SY5Y cells resistant to MPP-induced cytotoxicity. Furthermore, activation of the Nrf2 cascade by CRISPR/Cas9-induced Keap1 knockout protected SH-SY5Y cells from MPP. In Keap1 or PGK1 knockout SH-SY5Y cells,CBR-470-1 failed to offer further cytoprotection against MPP. Collectively PGK1 inhibition by CBR-470-1 protects SH-SY5Y cells from MPP via activation of the Keap1-Nrf2 cascade.
Topics: 1-Methyl-4-phenylpyridinium; Cell Line, Tumor; Gene Knockout Techniques; Humans; Kelch-Like ECH-Associated Protein 1; Mitochondria; NF-E2-Related Factor 2; Neuroprotective Agents; Neurotoxicity Syndromes; Phosphoglycerate Kinase; RNA, Small Interfering; Signal Transduction
PubMed: 32649311
DOI: 10.18632/aging.103443 -
FEBS Letters Feb 1989It has been shown that the denaturation of phosphoglycerate kinase (PGK) can be observed not only when the solution is heated above 30 degrees C, but also when it is...
It has been shown that the denaturation of phosphoglycerate kinase (PGK) can be observed not only when the solution is heated above 30 degrees C, but also when it is cooled below this temperature. The disruption of the native PGK structure upon cooling and the subsequent formation of this structure upon heating both proceed in two distinct stages which correspond to the independent disruption or reformation of each of its domains. In contrast, the heat denaturation of PGK proceeds in one stage, showing that the two domains of the molecule are associated into a single complex which figures in the denaturation process as a cooperative unit. It follows that, at elevated temperature, there is a positive interaction between the domains, which disappears at lower temperatures. This might be due to hydrophobic interactions, which are known to be temperature dependent. The temperature decrease leads to a decrease in inter- and intradomain interactions, which results in an increase of the independence of the domains and a decrease in their stability.
Topics: Cold Temperature; Hot Temperature; Phosphoglycerate Kinase; Protein Denaturation; Saccharomyces cerevisiae; Thermodynamics
PubMed: 2646151
DOI: 10.1016/0014-5793(89)80544-7 -
European Journal of Biochemistry Mar 1989Diffuse X-ray-scattering data give evidence for large-scale structural change in pig muscle 3-phosphoglycerate kinase upon substrate binding. Simultaneous binding of...
Diffuse X-ray-scattering data give evidence for large-scale structural change in pig muscle 3-phosphoglycerate kinase upon substrate binding. Simultaneous binding of 3-phosphoglycerate and MgATP either to the unmodified enzyme or to its active methylated derivative leads to about an 0.1-nm decrease in radius of gyration. These data coincide well with the previous data for yeast 3-phosphoglycerate kinase. When, instead of methylation, the two reactive thiol groups of pig muscle 3-phosphoglycerate kinase are carboxamidomethylated, the enzyme becomes inactive and the radii of gyration of its 'apo' and 'holo' forms do not differ within limits of experimental error. Thus, a correlation exists between the activity of 3-phosphoglycerate kinase and its substrate-induced large-scale conformational change. This correlation is a strong argument in favor of the functional importance of domain locking in the reaction catalyzed by 3-phosphoglycerate kinase.
Topics: Alkylation; Animals; Binding Sites; Methylation; Muscles; Phosphoglycerate Kinase; Protein Conformation; Scattering, Radiation; Substrate Specificity; Swine; X-Ray Diffraction; Yeasts
PubMed: 2707265
DOI: 10.1111/j.1432-1033.1989.tb14615.x -
Viruses Jun 2022The development of recessive resistance by loss of susceptibility is a consistent strategy to combat and limit damages caused by plant viruses. Susceptibility genes can... (Review)
Review
The development of recessive resistance by loss of susceptibility is a consistent strategy to combat and limit damages caused by plant viruses. Susceptibility genes can be turned into resistances, a feat that can either be selected among the plant's natural diversity or engineered by biotechnology. Here, we summarize the current knowledge on the phosphoglycerate kinases (PGK), which have emerged as a new class of susceptibility factors to single-stranded positive RNA viruses, including potyviruses. PGKs are metabolic enzymes involved in glycolysis and the carbon reduction cycle, encoded by small multigene families in plants. To fulfil their role in the chloroplast and in the cytosol, PGKs genes encode differentially addressed proteins. Here, we assess the diversity and homology of chloroplastic and cytosolic PGKs sequences in several crops and review the current knowledge on their redundancies during plant development, taking as a model. We also show how PGKs have been shown to be involved in susceptibility-and resistance-to viruses. Based on this knowledge, and drawing from the experience with the well-characterized translation initiation factors eIF4E, we discuss how PGKs genes, in light of their subcellular localization, function in metabolism, and susceptibility to viruses, could be turned into efficient genetic resistances using genome editing techniques.
Topics: Arabidopsis; Chloroplasts; Eukaryotic Initiation Factor-4E; Phosphoglycerate Kinase; Potyvirus
PubMed: 35746717
DOI: 10.3390/v14061245