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The FEBS Journal Mar 2015Cofactor-independent phosphoglycerate mutase (iPGM), an important enzyme in glycolysis and gluconeogenesis, catalyses the isomerization of 2- and 3-phosphoglycerates by...
Cofactor-independent phosphoglycerate mutase (iPGM), an important enzyme in glycolysis and gluconeogenesis, catalyses the isomerization of 2- and 3-phosphoglycerates by an Mn(2+)-dependent phospho-transfer mechanism via a phospho-enzyme intermediate. Crystal structures of bi-domain iPGM from Staphylococcus aureus, together with substrate-bound forms, have revealed a new conformation of the enzyme, representing an intermediate state of domain movement. The substrate-binding site and the catalytic site are present in two distinct domains in the intermediate form. X-ray crystallography complemented by simulated dynamics has enabled delineation of the complete catalytic cycle, which includes binding of the substrate, followed by its positioning into the catalytic site, phospho-transfer and finally product release. The present work describes a novel mechanism of domain movement controlled by a hydrophobic patch that is exposed on domain closure and acts like a spring to keep the protein in open conformation. Domain closing occurs after substrate binding, and is essential for phospho-transfer, whereas the open conformation is a prerequisite for efficient substrate binding and product dissociation. A new model of catalysis has been proposed by correlating the hinge-bending motion with the phospho-transfer mechanism.
Topics: Catalysis; Catalytic Domain; Computer Simulation; Crystallography, X-Ray; Ligands; Manganese; Models, Molecular; Motion; Phosphoglycerate Mutase; Protein Binding; Staphylococcus aureus; Substrate Specificity; Thermodynamics; X-Ray Diffraction
PubMed: 25611430
DOI: 10.1111/febs.13205 -
European Journal of Medicinal Chemistry Apr 2019Phosphoglycerate mutase 1 (PGAM1) coordinates glycolysis, pentose phosphate pathway, and serine synthesis to promote tumor growth through the regulation of its substrate...
Phosphoglycerate mutase 1 (PGAM1) coordinates glycolysis, pentose phosphate pathway, and serine synthesis to promote tumor growth through the regulation of its substrate 3-phosphoglycerate (3 PG) and product 2-phosphoglycerate (2 PG). Herein, based on our previously reported PGAM1 inhibitor PGMI-004A, we have developed anthraquinone derivatives as novel allosteric PGAM1 inhibitors and the structure-activity relationship (SAR) was investigated. In addition, we determined the co-crystal structure of PGAM1 and the inhibitor 8g, demonstrating that the inhibitor was located at a novel allosteric site. Among the derivatives, compound 8t was selected for further study, with IC values of 0.25 and approximately 5 μM in enzymatic and cell-based assays, respectively. Mechanistically, compound 8t reduced the glycolysis and oxygen consumption rate in cancer cells, which led to decreased adenosine 5'-triphosphate (ATP) production and subsequent 5' adenosine monophosphate-activated protein kinase (AMPK) activation. The inhibitor 8t also exhibited good efficacy in delaying tumor growth in H1299 xenograft model without obvious toxicity. Taken together, this proof-of-principle work further validates PGAM1 as a potential target for cancer therapy and provides useful information on anti-tumor drug discovery targeting PGAM1.
Topics: Animals; Anthraquinones; Antineoplastic Agents; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Screening Assays, Antitumor; Enzyme Inhibitors; Humans; Mice; Models, Molecular; Molecular Structure; Neoplasms, Experimental; Phosphoglycerate Mutase; Structure-Activity Relationship
PubMed: 30798052
DOI: 10.1016/j.ejmech.2019.01.085 -
Acta Biochimica Et Biophysica Sinica Aug 2023Tumor metabolic reprogramming and epigenetic modification work together to promote tumorigenesis and development. Protein lysine acetylation, which affects a variety of...
Tumor metabolic reprogramming and epigenetic modification work together to promote tumorigenesis and development. Protein lysine acetylation, which affects a variety of biological functions of proteins, plays an important role under physiological and pathological conditions. Here, through immunoprecipitation and mass spectrum data, we show that phosphoglycerate mutase 5 (PGAM5) deacetylation enhances malic enzyme 1 (ME1) metabolic enzyme activity to promote lipid synthesis and proliferation of liver cancer cells. Mechanistically, we demonstrate that the deacetylase SIRT2 mediates PGAM5 deacetylation to activate ME1 activity, leading to ME1 dephosphorylation, subsequent lipid accumulation and the proliferation of liver cancer cells. Taken together, our study establishes an important role for the SIRT2-PGAM5-ME1 axis in the proliferation of liver cancer cells, suggesting a potential innovative cancer therapy.
Topics: Humans; Sirtuin 2; Lipid Metabolism; Phosphoglycerate Mutase; Liver Neoplasms; Cell Proliferation; Lipids; Acetylation; Phosphoprotein Phosphatases; Mitochondrial Proteins
PubMed: 37580952
DOI: 10.3724/abbs.2023155 -
The Journal of Biological Chemistry Apr 2012Phosphoserine phosphatase (PSP) catalyzes the dephosphorylation of phosphoserine to serine and inorganic phosphate. PSPs, which have been found in all three domains of...
Phosphoserine phosphatase (PSP) catalyzes the dephosphorylation of phosphoserine to serine and inorganic phosphate. PSPs, which have been found in all three domains of life, belong to the haloacid dehalogenase-like hydrolase superfamily. However, certain organisms, particularly bacteria, lack a classical PSP gene, although they appear to possess a functional phosphoserine synthetic pathway. The apparent lack of a PSP ortholog in Hydrogenobacter thermophilus, an obligately chemolithoautotrophic and thermophilic bacterium, represented a missing link in serine anabolism because our previous study suggested that serine should be synthesized from phosphoserine. Here, we detected PSP activity in cell-free extracts of H. thermophilus and purified two proteins with PSP activity. Surprisingly, these proteins belonged to the histidine phosphatase superfamily and had been annotated as cofactor-dependent phosphoglycerate mutase (dPGM). However, because they possessed neither mutase activity nor the residues important for the activity, we defined these proteins as novel-type PSPs. Considering the strict substrate specificity toward l-phosphoserine, kinetic parameters, and PSP activity levels in cell-free extracts, these proteins were strongly suggested to function as PSPs in vivo. We also detected PSP activity from "dPGM-like" proteins of Thermus thermophilus and Arabidopsis thaliana, suggesting that PSP activity catalyzed by dPGM-like proteins may be distributed among a broad range of organisms. In fact, a number of bacterial genera, including Firmicutes and Cyanobacteria, were proposed to be strong candidates for possessing this novel type of PSP. These findings will help to identify the missing link in serine anabolism.
Topics: Amino Acid Sequence; Bacteria, Aerobic; Bacterial Proteins; Chromatography, Liquid; Coenzymes; Enzyme Assays; Kinetics; Molecular Weight; Phosphoglycerate Mutase; Phosphoric Monoester Hydrolases; Phylogeny; Protein Subunits; Recombinant Proteins; Sequence Analysis, Protein; Sequence Homology, Amino Acid; Substrate Specificity
PubMed: 22337887
DOI: 10.1074/jbc.M111.330621 -
European Journal of Biochemistry Feb 1995Spermatogenesis is a dramatic differentiation process which involves very selective but poorly characterized gene-expression patterns. To gain insight into this process,...
Spermatogenesis is a dramatic differentiation process which involves very selective but poorly characterized gene-expression patterns. To gain insight into this process, we have investigated the expression during spermatogenesis of the genes that encode phosphoglycerate mutase, an essential glycolytic enzyme for the spermatozoa energy supply. By using cDNA and genomic probes we demonstrate the presence in testis of a mRNA corresponding to the muscle-specific phosphoglycerate mutase which shows a longer poly(A) tail. This muscle-specific gene is submitted to developmental regulation during testis maturation and begins to be expressed at postnatal day 22, when germ cells start to enter into meiosis. Northern blot and in situ hybridization experiments show that in contrast to what happens during skeletal-muscle differentiation, PGAM-M gene expression during spermatogenesis is not coupled to constitutive phosphoglycerate mutase (PGAM-B) gene repression. Thus, the muscle-specific PGAM-M gene constitutes a meiotic gene and therefore represents a very interesting model to study differential tissue-specific gene expression.
Topics: Animals; Gene Expression Regulation, Developmental; Gene Expression Regulation, Enzymologic; In Situ Hybridization; Male; Muscle, Skeletal; Phosphoglycerate Mutase; Poly A; RNA, Messenger; Rats; Spermatogenesis; Testis; Tissue Distribution
PubMed: 7867621
DOI: 10.1111/j.1432-1033.1995.tb20182.x -
Life Sciences 1994Triiodothyronine (T3) increases phosphoglycerate mutase (PGAM) specific activity in rat skeletal and cardiac muscles. This increase is concomitant with an increase in...
Triiodothyronine (T3) increases phosphoglycerate mutase (PGAM) specific activity in rat skeletal and cardiac muscles. This increase is concomitant with an increase in the proportion of phosphoglycerate mutase isozymes which contain type-M subunit. Propylthiouracil (PTU), an anti-hormone, not only decreases phosphoglycerate mutase activity with respect to control rats, but also decreases the total M subunit contents. In liver, which only possesses type-B subunit phosphoglycerate mutase, none of the effects were detected.
Topics: Animals; Animals, Newborn; Drug Interactions; Electrophoresis; Heart; Isoenzymes; Liver; Muscles; Myocardium; Phosphoglycerate Mutase; Propylthiouracil; Rats; Rats, Sprague-Dawley; Spectrophotometry, Ultraviolet; Triiodothyronine
PubMed: 8107530
DOI: 10.1016/0024-3205(94)90003-5 -
The Journal of Biological Chemistry 2021Catalysis of human phosphoglycerate mutase is dependent on a 2,3-bisphosphoglycerate cofactor (dPGM), whereas the nonhomologous isozyme in many parasitic species is...
Catalysis of human phosphoglycerate mutase is dependent on a 2,3-bisphosphoglycerate cofactor (dPGM), whereas the nonhomologous isozyme in many parasitic species is cofactor independent (iPGM). This mechanistic and phylogenetic diversity offers an opportunity for selective pharmacologic targeting of glycolysis in disease-causing organisms. We previously discovered ipglycermide, a potent inhibitor of iPGM, from a large combinatorial cyclic peptide library. To fully delineate the ipglycermide pharmacophore, herein we construct a detailed structure-activity relationship using 280 substituted ipglycermide analogs. Binding affinities of these analogs to immobilized Caenorhabditis elegans iPGM, measured as fold enrichment relative to the index residue by deep sequencing of an mRNA display library, illuminated the significance of each amino acid to the pharmacophore. Using cocrystal structures and binding kinetics, we show that the high affinity of ipglycermide for iPGM orthologs, from Brugia malayi, Onchocerca volvulus, Dirofilaria immitis, and Escherichia coli, is achieved by a codependence between (1) the off-rate mediated by the macrocycle Cys14 thiolate coordination to an active-site Zn in the iPGM phosphatase domain and (2) shape complementarity surrounding the macrocyclic core at the phosphotransferase-phosphatase domain interface. Our results show that the high-affinity binding of ipglycermide to iPGMs freezes these structurally dynamic enzymes into an inactive, stable complex.
Topics: Animals; Catalytic Domain; Humans; Models, Molecular; Peptides, Cyclic; Phosphoglycerate Mutase; Phylogeny; Protein Conformation; Structure-Activity Relationship
PubMed: 33812994
DOI: 10.1016/j.jbc.2021.100628 -
The Journal of Cell Biology Mar 2014Despite the well-documented clinical significance of the Warburg effect, it remains unclear how the aggressive glycolytic rates of tumor cells might contribute to other...
Despite the well-documented clinical significance of the Warburg effect, it remains unclear how the aggressive glycolytic rates of tumor cells might contribute to other hallmarks of cancer, such as bypass of senescence. Here, we report that, during oncogene- or DNA damage-induced senescence, Pak1-mediated phosphorylation of phosphoglycerate mutase (PGAM) predisposes the glycolytic enzyme to ubiquitin-mediated degradation. We identify Mdm2 as a direct binding partner and ubiquitin ligase for PGAM in cultured cells and in vitro. Mutations in PGAM and Mdm2 that abrogate ubiquitination of PGAM restored the proliferative potential of primary cells under stress conditions and promoted neoplastic transformation. We propose that Mdm2, a downstream effector of p53, attenuates the Warburg effect via ubiquitination and degradation of PGAM.
Topics: Animals; Cell Line; Cellular Senescence; DNA Damage; Down-Regulation; HCT116 Cells; HEK293 Cells; HT29 Cells; HeLa Cells; Humans; Mice; Mice, Inbred C57BL; Mice, Nude; Phosphoglycerate Mutase; Phosphorylation; Proteolysis; Proto-Oncogene Proteins c-mdm2; Stress, Physiological; Ubiquitin; p21-Activated Kinases
PubMed: 24567357
DOI: 10.1083/jcb.201306149 -
The Journal of Biological Chemistry Feb 2012Emerging proteomic evidence suggests that acetylation of metabolic enzymes is a prevalent post-translational modification. In a few recent reports, acetylation...
Emerging proteomic evidence suggests that acetylation of metabolic enzymes is a prevalent post-translational modification. In a few recent reports, acetylation down-regulated activity of specific enzymes in fatty acid oxidation, urea cycle, electron transport, and anti-oxidant pathways. Here, we reveal that the glycolytic enzyme phosphoglycerate mutase-1 (PGAM1) is negatively regulated by Sirt1, a member of the NAD(+)-dependent protein deacetylases. Acetylated PGAM1 displays enhanced activity, although Sirt1-mediated deacetylation reduces activity. Acetylation sites mapped to the C-terminal "cap," a region previously known to affect catalytic efficiency. Overexpression of a constitutively active variant (acetylated mimic) of PGAM1 stimulated flux through glycolysis. Under glucose restriction, Sirt1 levels dramatically increased, leading to PGAM1 deacetylation and attenuated activity. Previously, Sirt1 has been implicated in the adaptation from glucose to fat burning. This study (i) demonstrates that protein acetylation can stimulate metabolic enzymes, (ii) provides biochemical evidence that glycolysis is modulated by reversible acetylation, and (iii) demonstrates that PGAM1 deacetylation and activity are directly controlled by Sirt1.
Topics: Acetylation; Glycolysis; HEK293 Cells; Humans; Phosphoglycerate Mutase; Sirtuin 1
PubMed: 22157007
DOI: 10.1074/jbc.M111.317404 -
Oncology Reports Oct 2016Previous studies indicated that phosphoglycerate mutase 1 (PGAM1) is involved in many cancer types and promotes breast cancer progression. However, the role of PGAM1 in...
Previous studies indicated that phosphoglycerate mutase 1 (PGAM1) is involved in many cancer types and promotes breast cancer progression. However, the role of PGAM1 in glioma remains unclear. The present study aimed to investigate the association of PGAM1 expression with glioma grade and the role of PGAM1 in proliferation, apoptosis, migration and invasion of glioma cells. The mRNA and protein expression of PGAM1 was analysed in glioma tissues and normal brain tissues. The expression of PGAM1 was examined further by immunohistochemical analysis. In addition, we inhibited the expression of PGAM1 in glioma cell line by siRNA to evaluate its role in glioma proliferation, apoptosis, migration and invasion. The mRNA and protein expression of PGAM1 was significantly greater in glioma than normal brain tissues. PGAM1 expression was associated with the WHO grade of glioma. siRNA knockdown of PGAM1 significantly inhibited glioma cell proliferation, promoted glioma cell apoptosis, induced S phase cell cycle arrest and inhibited glioma cell migration and invasion in vitro. PGAM1 may be associated with the grade of glioma and be involved in the biological behavior of glioma cells. PGAM1 might be a novel therapeutic target in glioma.
Topics: Adult; Aged; Apoptosis; Brain; Cell Cycle; Cell Movement; Cell Proliferation; Female; Gene Expression Regulation, Neoplastic; Glioma; Humans; Male; Middle Aged; Neoplasm Invasiveness; Phosphoglycerate Mutase; RNA, Messenger; RNA, Small Interfering
PubMed: 27572934
DOI: 10.3892/or.2016.5046