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Advances in Protein Chemistry and... 2017Enzymes in the α-d-phosphohexomutases superfamily catalyze the reversible conversion of phosphosugars, such as glucose 1-phosphate and glucose 6-phosphate. These...
Enzymes in the α-d-phosphohexomutases superfamily catalyze the reversible conversion of phosphosugars, such as glucose 1-phosphate and glucose 6-phosphate. These reactions are fundamental to primary metabolism across the kingdoms of life and are required for a myriad of cellular processes, ranging from exopolysaccharide production to protein glycosylation. The subject of extensive mechanistic characterization during the latter half of the 20th century, these enzymes have recently benefitted from biophysical characterization, including X-ray crystallography, NMR, and hydrogen-deuterium exchange studies. This work has provided new insights into the unique catalytic mechanism of the superfamily, shed light on the molecular determinants of ligand recognition, and revealed the evolutionary conservation of conformational flexibility. Novel associations with inherited metabolic disease and the pathogenesis of bacterial infections have emerged, spurring renewed interest in the long-appreciated functional roles of these enzymes.
Topics: Amino Acid Sequence; Animals; Bacteria; Bacterial Infections; Catalytic Domain; Crystallography, X-Ray; Glucosephosphates; Humans; Metabolic Diseases; Mutation; Nuclear Magnetic Resonance, Biomolecular; Phosphoglucomutase; Protein Conformation; Sequence Alignment
PubMed: 28683921
DOI: 10.1016/bs.apcsb.2017.04.005 -
Proceedings of the National Academy of... Jan 1992A cDNA clone encoding the mRNA for the highly polymorphic human enzyme phosphoglucomutase 1 (PGM1; EC 5.4.2.2) has been isolated and characterized. This was achieved... (Comparative Study)
Comparative Study
A cDNA clone encoding the mRNA for the highly polymorphic human enzyme phosphoglucomutase 1 (PGM1; EC 5.4.2.2) has been isolated and characterized. This was achieved indirectly by first isolating a rabbit cDNA from an expression library using anti-rabbit PGM antibodies. A comparison of the nucleotide sequences shows that the homologies between human and rabbit PGM1 mRNAs are 92% and 97% for the coding nucleotide sequence and the amino acid sequence, respectively. The derived rabbit amino acid sequence is in complete agreement with the published protein sequence for rabbit muscle PGM. A physical localization of the human PGM1 gene to chromosome 1p31 has been determined by in situ hybridization. Analysis of DNA from a wide variety of vertebrates indicates a high level of PGM1 sequence conservation during evolution.
Topics: Animals; Base Sequence; Blotting, Southern; Chromosome Banding; Chromosome Mapping; Chromosomes, Human, Pair 1; Cloning, Molecular; DNA; Humans; Molecular Sequence Data; Nucleic Acid Hybridization; Oligodeoxyribonucleotides; Phosphoglucomutase; Polymerase Chain Reaction; RNA, Messenger; Rabbits; Sequence Alignment
PubMed: 1530890
DOI: 10.1073/pnas.89.1.411 -
Scientific Reports Jun 2020In solid tumors, hypoxia can trigger aberrant expression of transcription factors and genes, resulting in abnormal biological functions such as altered energetic...
In solid tumors, hypoxia can trigger aberrant expression of transcription factors and genes, resulting in abnormal biological functions such as altered energetic pathways in cancer cells. Glucose metabolism is an important part of this phenomenon, which is associated with changes in the functional expression of transporters and enzymes involved in the glycolysis pathway. The latter phenomenon can finally lead to the lactate accumulation and pH dysregulation in the tumor microenvironment and subsequently further invasion and metastasis of cancer cells. Having capitalized on the computational modeling, in this study, for the first time, we aimed to investigate the effects of hypoxia-induced factor-1 (HIF-1) mediated hypoxia on the magnitude of functional expression of all the enzymes and transporters involved in the glycolysis process. The main objective was to establish a quantitative relationship between the hypoxia intensity and the intracellular lactate levels and determine the key regulators of the glycolysis pathway. This model clearly showed an increase in the lactate concentration during the oxygen depletion. The proposed model also predicted that the phosphofructokinase-1 and phosphoglucomutase enzymes might play the most important roles in the regulation of the lactate production.
Topics: Gene Expression; Glycolysis; Humans; Hypoxia; Hypoxia-Inducible Factor 1, alpha Subunit; Lactic Acid; Models, Theoretical; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasms; Phosphofructokinase-1; Phosphoglucomutase; Signal Transduction; Tumor Microenvironment
PubMed: 32514127
DOI: 10.1038/s41598-020-66059-w -
The Journal of Allergy and Clinical... May 2014Identifying genetic syndromes that lead to significant atopic disease can open new pathways for investigation and intervention in allergy. (Clinical Trial)
Clinical Trial
BACKGROUND
Identifying genetic syndromes that lead to significant atopic disease can open new pathways for investigation and intervention in allergy.
OBJECTIVE
We sought to define a genetic syndrome of severe atopy, increased serum IgE levels, immune deficiency, autoimmunity, and motor and neurocognitive impairment.
METHODS
Eight patients from 2 families with similar syndromic features were studied. Thorough clinical evaluations, including brain magnetic resonance imaging and sensory evoked potentials, were performed. Peripheral lymphocyte flow cytometry, antibody responses, and T-cell cytokine production were measured. Whole-exome sequencing was performed to identify disease-causing mutations. Immunoblotting, quantitative RT-PCR, enzymatic assays, nucleotide sugar, and sugar phosphate analyses, along with matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry of glycans, were used to determine the molecular consequences of the mutations.
RESULTS
Marked atopy and autoimmunity were associated with increased T(H)2 and T(H)17 cytokine production by CD4(+) T cells. Bacterial and viral infection susceptibility were noted along with T-cell lymphopenia, particularly of CD8(+) T cells, and reduced memory B-cell numbers. Apparent brain hypomyelination resulted in markedly delayed evoked potentials and likely contributed to neurologic abnormalities. Disease segregated with novel autosomal recessive mutations in a single gene, phosphoglucomutase 3 (PGM3). Although PGM3 protein expression was variably diminished, impaired function was demonstrated by decreased enzyme activity and reduced uridine diphosphate-N-acetyl-D-glucosamine, along with decreased O- and N-linked protein glycosylation in patients' cells. These results define a new congenital disorder of glycosylation.
CONCLUSIONS
Autosomal recessive hypomorphic PGM3 mutations underlie a disorder of severe atopy, immune deficiency, autoimmunity, intellectual disability, and hypomyelination.
Topics: Autoimmune Diseases; B-Lymphocytes; CD8-Positive T-Lymphocytes; Child; Child, Preschool; Cognition Disorders; Common Variable Immunodeficiency; Family; Female; Genetic Diseases, Inborn; Humans; Hypersensitivity; Immunoglobulin E; Male; Mutation; Pedigree; Phosphoglucomutase; Th17 Cells; Th2 Cells; Young Adult
PubMed: 24589341
DOI: 10.1016/j.jaci.2014.02.013 -
The Journal of Biological Chemistry Oct 2001Lithium is a drug frequently used in the treatment of manic depressive disorder. We have observed that the yeast Saccharomyces cerevisiae is very sensitive to lithium...
Lithium is a drug frequently used in the treatment of manic depressive disorder. We have observed that the yeast Saccharomyces cerevisiae is very sensitive to lithium when growing in galactose medium. In this work we show that lithium inhibits with high affinity yeast (IC50 approximately 0.2 mm) and human (IC50 approximately 1.5 mm) phosphoglucomutase, the enzyme that catalyzes the reversible conversion of glucose 1-phosphate to glucose 6-phosphate. Lithium inhibits the rate of fermentation when yeast are grown in galactose and induces accumulation of glucose 1-phosphate and galactose 1-phosphate. Accumulation of these metabolites was also observed when a strain deleted of the two isoforms of phosphoglucomutase was incubated in galactose medium. In glucose-grown cells lithium reduces the steady state levels of UDP-glucose, resulting in a defect on trehalose and glycogen biosynthesis. Lithium acts as a competitive inhibitor of yeast phosphoglucomutase activity by competing with magnesium, a cofactor of the enzyme. High magnesium concentrations revert lithium inhibition of growth and phosphoglucomutase activity. Lithium stress causes an increase of the phosphoglucomutase activity due to an induction of transcription of the PGM2 gene, and its overexpression confers lithium tolerance in galactose medium. These results show that phosphoglucomutase is an important in vivo lithium target.
Topics: Blotting, Northern; Cell Line; Cell-Free System; Culture Media; Fermentation; Galactose; Glucose; Humans; Lithium; Magnesium; Phosphoglucomutase; Saccharomyces cerevisiae
PubMed: 11500487
DOI: 10.1074/jbc.M101451200 -
The Journal of Biological Chemistry Nov 2021Cyclic-di-adenosine monophosphate (c-di-AMP) is an important nucleotide signaling molecule that plays a key role in osmotic regulation in bacteria. c-di-AMP is produced...
Cyclic-di-adenosine monophosphate (c-di-AMP) is an important nucleotide signaling molecule that plays a key role in osmotic regulation in bacteria. c-di-AMP is produced from two molecules of ATP by proteins containing a diadenylate cyclase (DAC) domain. In Bacillus subtilis, the main c-di-AMP cyclase, CdaA, is a membrane-linked cyclase with an N-terminal transmembrane domain followed by the cytoplasmic DAC domain. As both high and low levels of c-di-AMP have a negative impact on bacterial growth, the cellular levels of this signaling nucleotide are tightly regulated. Here we investigated how the activity of the B. subtilis CdaA is regulated by the phosphoglucomutase GlmM, which has been shown to interact with the c-di-AMP cyclase. Using the soluble B. subtilis CdaA catalytic domain and purified full-length GlmM or the GlmM variant lacking the C-terminal flexible domain 4, we show that the cyclase and phosphoglucomutase form a stable complex in vitro and that GlmM is a potent cyclase inhibitor. We determined the crystal structure of the individual B. subtilis CdaA and GlmM homodimers and of the CdaA:GlmM complex. In the complex structure, a CdaA dimer is bound to a GlmM dimer in such a manner that GlmM blocks the oligomerization of CdaA and formation of active head-to-head cyclase oligomers, thus suggesting a mechanism by which GlmM acts as a cyclase inhibitor. As the amino acids at the CdaA:GlmM interphase are conserved, we propose that the observed mechanism of inhibition of CdaA by GlmM may also be conserved among Firmicutes.
Topics: Bacillus subtilis; Bacterial Proteins; Crystallography, X-Ray; Multienzyme Complexes; Phosphoglucomutase; Phosphorus-Oxygen Lyases; Protein Domains; Protein Multimerization; Protein Structure, Quaternary
PubMed: 34678313
DOI: 10.1016/j.jbc.2021.101317 -
Plant Physiology Dec 2010Cytosolic phosphoglucomutase (cPGM) interconverts glucose-6-phosphate and glucose-1-phosphate and is a key enzyme of central metabolism. In this study, we show that...
Cytosolic phosphoglucomutase (cPGM) interconverts glucose-6-phosphate and glucose-1-phosphate and is a key enzyme of central metabolism. In this study, we show that Arabidopsis (Arabidopsis thaliana) has two cPGM genes (PGM2 and PGM3) encoding proteins with high sequence similarity and redundant functions. Whereas pgm2 and pgm3 single mutants were undistinguishable from the wild type, loss of both PGM2 and PGM3 severely impaired male and female gametophyte function. Double mutant pollen completed development but failed to germinate. Double mutant ovules also developed normally, but approximately half remained unfertilized 2 d after pollination. We attribute these phenotypes to an inability to effectively distribute carbohydrate from imported or stored substrates (e.g. sucrose) into the major biosynthetic (e.g. cell wall biosynthesis) and respiratory pathways (e.g. glycolysis and the oxidative pentose phosphate pathway). Disturbing these pathways is expected to have dramatic consequences for germinating pollen grains, which have high metabolic and biosynthetic activities. We propose that residual cPGM mRNA or protein derived from the diploid mother plant is sufficient to enable double mutant female gametophytes to attain maturity and for some to be fertilized. Mature plants possessing a single cPGM allele had a major reduction in cPGM activity. However, photosynthetic metabolism and growth were normal, suggesting that under standard laboratory conditions cPGM activity provided from one wild-type allele is sufficient to mediate the photosynthetic and respiratory fluxes in leaves.
Topics: Arabidopsis; Cytosol; Genes, Plant; Germ Cells, Plant; Germination; Mutation; Phosphoglucomutase; Phylogeny; Pollen
PubMed: 20959421
DOI: 10.1104/pp.110.165027 -
FEBS Letters Aug 2014Phosphoglucomutase (PGM)1 catalyzes the reversible conversion reaction between glucose-1-phosphate (G-1-P) and glucose-6-phosphate (G-6-P). Although both G-1-P and G-6-P...
Phosphoglucomutase (PGM)1 catalyzes the reversible conversion reaction between glucose-1-phosphate (G-1-P) and glucose-6-phosphate (G-6-P). Although both G-1-P and G-6-P are important intermediates for glucose and glycogen metabolism, the biological roles and regulatory mechanisms of PGM1 are largely unknown. In this study we found that T553 is obligatory for PGM1 stability and the last C-terminal residue, T562, is critical for its activity. Interestingly, depletion of PGM1 was associated with declined cellular glycogen content and decreased rates of glycogenolysis and glycogenesis. Furthermore, PGM1 depletion suppressed cell proliferation under long-term repetitive glucose depletion. Our results suggest that PGM1 is required for sustained cell growth during nutritional changes, probably through regulating the balance of G-1-P and G-6-P in order to satisfy the cellular demands during nutritional stress.
Topics: Amino Acid Sequence; Animals; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Enzyme Stability; Glucose; Glucose-6-Phosphate; Glucosephosphates; Glycogen; Humans; Phosphoglucomutase; Threonine
PubMed: 24952355
DOI: 10.1016/j.febslet.2014.06.034 -
Journal of Bacteriology Jun 2021
Topics: Bacterial Capsules; Biosynthetic Pathways; Phosphoglucomutase; Streptococcus pneumoniae
PubMed: 34100630
DOI: 10.1128/JB.00220-21 -
Journal of Bacteriology Feb 1973Two very poorly lytic mutants of Bacillus licheniformis 6346 that had no teichuronic acid or glucose in their walls were phosphoglucomutase deficient. The walls of the...
Two very poorly lytic mutants of Bacillus licheniformis 6346 that had no teichuronic acid or glucose in their walls were phosphoglucomutase deficient. The walls of the mutants were less autolytic, and the lesion in the phosphoglucomutase gene and the formation of lytic amidase seemed to be interrelated. When phosphoglucomutase was regained or the effects of the deficiency were circumvented by the presence of galactose in the medium, the lytic enzyme was partially regained. When subjected to growth limitation by the supply of inorganic phosphate, the mutants ceased to make teichoic acid, and their walls contained a greatly increased proportion of mucopeptide. Under these conditions they formed irregular spheres which changed back to rods when inorganic phosphate was supplied. Both wall and protein synthesis were necessary for the changes in morphology. An intermediate crescent-shaped cell was formed in the change from sphere to a rod. The possible relationship of this morphological change to the distribution of biosynthetic sites is discussed.
Topics: Amidohydrolases; Autolysis; Bacillus; Bacteriological Techniques; Carbohydrates; Cell Wall; Cell-Free System; Culture Media; Galactose; Glucose; Glycerol; Magnesium; Microscopy, Electron; Microscopy, Phase-Contrast; Mutation; Nucleotidyltransferases; Phosphates; Phosphoglucomutase; Teichoic Acids; Uronic Acids
PubMed: 4570613
DOI: 10.1128/jb.113.2.969-984.1973