-
FEBS Letters Mar 1974
Comparative Study Review
Topics: Alcohol Oxidoreductases; Carbohydrate Epimerases; Fructose-Bisphosphatase; Fructose-Bisphosphate Aldolase; Fructosephosphates; Glucose; Glucose Oxidase; Glucose-6-Phosphatase; Glucose-6-Phosphate Isomerase; Glucosephosphate Dehydrogenase; Glucosephosphates; Hexokinase; Hexosediphosphates; Humans; Kinetics; Organ Specificity; Phosphoglucomutase; Phosphotransferases; Species Specificity; Structure-Activity Relationship
PubMed: 4368416
DOI: 10.1016/0014-5793(74)80693-9 -
Genes Nov 2022Sweet potato (Ipomoea batatas), an important root crop, has storage roots rich in starch that are edible and serve as a raw material in bioenergy production. Increasing...
Sweet potato (Ipomoea batatas), an important root crop, has storage roots rich in starch that are edible and serve as a raw material in bioenergy production. Increasing the storage-root starch contents is a key sweet potato breeding goal. Phosphoglucomutase (PGM) is the catalytic enzyme for the interconversion of glucose-6-phosphate and glucose-1-phosphate, precursors in the plant starch synthetic pathway. Plant PGMs have plastidial and cytosolic isoforms, based on their subcellular localization. Here, , containing 22 exons and 21 introns, was cloned from the sweet potato line Xu 781. This gene was highly expressed in the storage roots and leaves, and its expression was induced by exogenous sucrose treatments. The mature IbpPGM protein was successfully expressed in when a 73-aa chloroplastic transit peptide detected in the N-terminus was excised. The subcellular localization confirmed that IbpPGM was localized to the chloroplasts. The low-starch sweet potato cultivar Lizixiang -overexpression lines showed significantly increased starch, glucose, and fructose levels but a decreased sucrose level. Additionally, the expression levels of the starch synthetic pathway genes in the storage roots were up-regulated to different extents. Thus, significantly increased the starch content of the sweet potato storage roots, which makes it a candidate gene for the genetic engineering of the sweet potato.
Topics: Starch; Ipomoea batatas; Phosphoglucomutase; Plant Roots; Plant Breeding; Sucrose
PubMed: 36553501
DOI: 10.3390/genes13122234 -
ELife Oct 2022The most common cause of human congenital disorders of glycosylation (CDG) are mutations in the phosphomannomutase gene which affect protein -linked glycosylation. The...
The most common cause of human congenital disorders of glycosylation (CDG) are mutations in the phosphomannomutase gene which affect protein -linked glycosylation. The yeast gene encodes a homolog of human . We evolved 384 populations of yeast harboring one of two human-disease-associated alleles, V238M and -F126L, or wild-type . We find that after 1000 generations, most populations compensate for the slow-growth phenotype associated with the human-disease-associated alleles. Through whole-genome sequencing we identify compensatory mutations, including known genetic interactors. We observe an enrichment of compensatory mutations in other genes whose human homologs are associated with Type 1 CDG, including , which encodes the minor isoform of phosphoglucomutase in yeast. By genetic reconstruction, we show that evolved mutations are dominant and allele-specific genetic interactors that restore both protein glycosylation and growth of yeast harboring the -V238M allele. Finally, we characterize the enzymatic activity of purified Pgm1 mutant proteins. We find that reduction, but not elimination, of Pgm1 activity best compensates for the deleterious phenotypes associated with the -V238M allele. Broadly, our results demonstrate the power of experimental evolution as a tool for identifying genes and pathways that compensate for human-disease-associated alleles.
Topics: Humans; Saccharomyces cerevisiae; Congenital Disorders of Glycosylation; Phosphoglucomutase; Mutant Proteins; Saccharomyces cerevisiae Proteins
PubMed: 36214454
DOI: 10.7554/eLife.79346 -
Molecules and Cells Feb 2014A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A...
A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-α-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the 5(th) hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-β-D-glucoside, genistein 7-O-β-D-glucoside (genistin), genistein 4',7-O-β-D-diglucoside, biochanin A-7-O-β-D-glucoside (sissotrin), daidzein 4'-O-β-D-glucoside, daidzein 7-O-β-D-glucoside (daidzin), daidzein 4', 7-O-β-D-diglucoside, and formononetin 7-O-β-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOFESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli BL21(DE3)/ΔpgiΔzwfΔushA overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of 200 μM of supplemented isoflavonoids, without any additional UDP-α-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.
Topics: Bacillus; Bacterial Proteins; Chromatography, Liquid; Escherichia coli; Escherichia coli Proteins; Fermentation; Glucosides; Glycosylation; Glycosyltransferases; Isoflavones; Phosphoglucomutase; Spectrometry, Mass, Electrospray Ionization; UTP-Glucose-1-Phosphate Uridylyltransferase
PubMed: 24599002
DOI: 10.14348/molcells.2014.2348 -
MBio Aug 2022The reactions of α-d-phosphohexomutases (αPHM) are ubiquitous, key to primary metabolism, and essential for several processes in all domains of life. The functionality...
The reactions of α-d-phosphohexomutases (αPHM) are ubiquitous, key to primary metabolism, and essential for several processes in all domains of life. The functionality of these enzymes relies on an initial phosphorylation step which requires the presence of α-d-glucose-1,6-bisphosphate (Glc-1,6-BP). While well investigated in vertebrates, the origin of this activator compound in bacteria is unknown. Here we show that the Slr1334 protein from the unicellular cyanobacterium sp. PCC 6803 is a Glc-1,6-BP-synthase. Biochemical analysis revealed that Slr1334 efficiently converts fructose-1,6-bisphosphate (Frc-1,6-BP) and α-d-glucose-1-phosphate/α-d-glucose-6-phosphate into Glc-1,6-BP and also catalyzes the reverse reaction. As inferred from phylogenetic analysis, the product belongs to a primordial subfamily of αPHMs that is present especially in deeply branching bacteria and also includes human commensals and pathogens. Remarkably, the homologue of Slr1334 in the human gut bacterium Bacteroides salyersiae catalyzes the same reaction, suggesting a conserved and essential role for the members of this αPHM subfamily. Glc-1,6-BP is known as an essential activator of phosphoglucomutase (PGM) and other members of the αPHM superfamily, making it a central regulator in glycogen metabolism, glycolysis, amino sugar formation as well as bacterial cell wall and capsule formation. Despite this essential role in carbon metabolism, its origin in prokaryotes has so far remained elusive. In this study we identify a member of a specific αPHM subfamily as the first bacterial Glc-1,6-BP synthase, forming free Glc-1,6-BP by using Frc-1,6-BP as phosphoryl-donor. PGMs of this subfamily are widely distributed among prokaryotes including human commensals and pathogens. By showing that a distinct subfamily member can also form Glc-1,6-BP, we provide evidence that Glc-1,6-BP synthase activity is a general feature of this group.
Topics: Animals; Glucose; Glucose-6-Phosphate; Humans; Phosphoglucomutase; Phylogeny
PubMed: 35856562
DOI: 10.1128/mbio.01469-22 -
International Journal of Molecular... Aug 2020For decades, lithium chloride (LiCl) has been used as a treatment option for those living with bipolar disorder (BD). As a result, many studies have been conducted to...
For decades, lithium chloride (LiCl) has been used as a treatment option for those living with bipolar disorder (BD). As a result, many studies have been conducted to examine its mode of action, toxicity, and downstream cellular responses. We know that LiCl is able to affect cell signaling and signaling transduction pathways through protein kinase C and glycogen synthase kinase-3, which are considered to be important in regulating gene expression at the translational level. However, additional downstream effects require further investigation, especially in translation pathway. In yeast, LiCl treatment affects the expression, and thus the activity, of , a phosphoglucomutase involved in sugar metabolism. Inhibition of leads to the accumulation of intermediate metabolites of galactose metabolism causing cell toxicity. However, it is not fully understood how LiCl affects gene expression in this matter. In this study, we identified three genes, , , and , which increase yeast LiCl sensitivity when deleted. We further demonstrate that , , and influence translation and exert their activity through the 5'-Untranslated region (5'-UTR) of mRNA in yeast.
Topics: 5' Untranslated Regions; Amino Acyl-tRNA Synthetases; Antimanic Agents; Bipolar Disorder; Gene Expression Regulation; Gene Knockout Techniques; Lithium Chloride; Organisms, Genetically Modified; Phosphoglucomutase; Protein Biosynthesis; RNA Helicases; RNA, Messenger; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Signal Transduction
PubMed: 32785068
DOI: 10.3390/ijms21165730 -
Journal of Microbiology and... May 2019β-Glucan is a chief structural polymer in the cell wall of yeast. β-Glucan has attracted intensive attention because of its wide applications in health protection and...
β-Glucan is a chief structural polymer in the cell wall of yeast. β-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the β-glucan biosynthesis pathway in was engineered to enhance β-glucan accumulation. A newly identified bacterial β-1, 6-glucan synthase GsmA from was expressed, and increased β-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of β-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased β-glucan accumulation. After further optimization of culture conditions, the β-glucan content was increased by 53.1%. This study provides a new approach to enhance β-glucan biosynthesis in .
Topics: Biosynthetic Pathways; Carbohydrate Metabolism; Cell Wall; Culture Media; Glucans; Glucosyltransferases; Metabolic Engineering; Mycoplasma agalactiae; Phosphoglucomutase; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; beta-Glucans; rho GTP-Binding Proteins
PubMed: 30955255
DOI: 10.4014/jmb.1812.12049 -
The Journal of Biological Chemistry Sep 1964
Topics: Amino Acids; Animals; Benzoates; Cysteine; Enzyme Inhibitors; Escherichia coli; Galactose; Glucose; Glycerol; Imidazoles; Magnesium; Muscles; Phosphates; Phosphoglucomutase; Rabbits; Research; Succinates; Sulfhydryl Compounds
PubMed: 14216423
DOI: No ID Found -
Biochemistry Jul 2018α-Phosphoglucomutase (αPGM), in its phosphorylated state, catalyzes the interconversion of α-d-glucose 1-phosphate and α-d-glucose 6-phosphate. The αPGM of...
α-Phosphoglucomutase (αPGM), in its phosphorylated state, catalyzes the interconversion of α-d-glucose 1-phosphate and α-d-glucose 6-phosphate. The αPGM of Lactococcus lactis is a type C2B member of the haloalkanoic acid dehalogenase (HAD) enzyme family and is comprised of a Rossmann-fold catalytic domain and inserted α/β-fold cap domain. The active site is formed at the domain-domain interface. Herein, we report the results from a kinetic-based study of L. lactis αPGM catalysis, which demonstrate enzyme activation by autocatalyzed phosphorylation of Asp8 with αG1P, the intermediacy of αG1,6bisP in the phospho Ll-αPGM-catalyzed conversion of αG1P to G6P, and the reorientation of the αG1,6bisP intermediate via dissociation to solvent and rebinding. In order to provide insight into the structural determinants of L. lactis αPGM substrate recognition and catalysis, metal cofactor and substrate specificities were determined as were the contributions made by active-site residues toward catalytic efficiency. Lastly, the structure and catalytic mechanism of L. lactis αPGM are compared with those of HAD family phosphomutases L. lactis β-phosphoglucomutase and eukayotic α-phosphomannomutase to provide insight into the evolution of phosphohexomutases from HAD family phosphatases.
Topics: Catalytic Domain; Crystallography, X-Ray; Enzyme Activation; Glucose-6-Phosphate; Glucosephosphates; Kinetics; Lactococcus lactis; Models, Molecular; Phosphoglucomutase; Phosphorylation; Protein Conformation; Substrate Specificity
PubMed: 29952545
DOI: 10.1021/acs.biochem.8b00396 -
Molecular Microbiology Aug 2012The enzymes phosphomannomutase (PMM), phospho-N-acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between the C6...
The enzymes phosphomannomutase (PMM), phospho-N-acetylglucosamine mutase (PAGM) and phosphoglucomutase (PGM) reversibly catalyse the transfer of phosphate between the C6 and C1 hydroxyl groups of mannose, N-acetylglucosamine and glucose respectively. Although genes for a candidate PMM and a PAGM enzymes have been found in the Trypanosoma brucei genome, there is, surprisingly, no candidate gene for PGM. The TbPMM and TbPAGM genes were cloned and expressed in Escherichia coli and the TbPMM enzyme was crystallized and its structure solved at 1.85 Å resolution. Antibodies to the recombinant proteins localized endogenous TbPMM to glycosomes in the bloodstream form of the parasite, while TbPAGM localized to both the cytosol and glycosomes. Both recombinant enzymes were able to interconvert glucose-phosphates, as well as acting on their own definitive substrates. Analysis of sugar nucleotide levels in parasites with TbPMM or TbPAGM knocked down by RNA interference (RNAi) suggests that, in vivo, PGM activity is catalysed by both enzymes. This is the first example in any organism of PGM activity being completely replaced in this way and it explains why, uniquely, T. brucei has been able to lose its PGM gene. The RNAi data for TbPMM also showed that this is an essential gene for parasite growth.
Topics: Acetylglucosamine; Amino Acid Motifs; Amino Acid Sequence; Glucose-6-Phosphate; Glucosephosphates; Kinetics; Mannosephosphates; Models, Molecular; Molecular Sequence Data; Open Reading Frames; Phosphoglucomutase; Phosphotransferases (Phosphomutases); Protein Conformation; Protein Transport; RNA Interference; Recombinant Proteins; Sequence Alignment; Trypanosoma brucei brucei
PubMed: 22676716
DOI: 10.1111/j.1365-2958.2012.08124.x