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Influence of bradykinin on glucose 1,6-bisphosphate and cyclic GMP levels and on the activities of glucose 1,6-bisphosphatase, phosphofructokinase and phosphoglucomutase in muscle.

The intracellular concentration of glucose-1,6-bisphosphate (Glc-1,6-P2) in rat tibialis anterior muscle was markedly decreased following the injection of bradykinin. Injection of bradykinin also induced a significant increase in the level of cyclic GMP in muscle. The activity of glucose-1,6-bisphosphatase, the enzyme that degrades Glc-1,6-P2, was markedly enhanced by bradykinin, which may account for the decrease in the level of Glc-1,6-P2. The decrease in Glc-1,6-P2, the potent activator of phosphofructokinase and phosphoglucomutase, was accompanied by a concomitant reduction in these enzymes' activities. The bradykinin-induced decrease in Glc-1,6-P2 and in the activity of phosphofructokinase, the rate-limiting enzyme in glycolysis, may be involved in the pathogenic influences of this hormone in various clinical conditions.

Animals↗

Trifluoperazine abolishes the actions of bradykinin on glucose 1,6-bisphosphate levels and on the activities of glucose 1,6-bisphosphatase, phosphofructokinase and phosphoglucomutase.

Injection of trifluoperazine abolished the bradykinin-induced decrease in intracellular concentration of glucose 1,6-bisphosphate (Glc-1,6-P2) in rat tibialis anterior muscle and skin. These changes in Glc-1,6-P2 levels may be attributed to the changes in the activity of glucose 1,6-bisphosphatase (the enzyme that degrades Glc-1,6-P2), which was markedly enhanced by bradykinin and reversed by trifluoperazine. Concomitantly to the changes in Glc-1,6-P2, the potent activator of phosphofructokinase and phosphoglucomutase, the activities of these enzymes were reduced by bradykinin and restored by trifluoperazine. These findings suggest that trifluoperazine treatment may have a beneficial effect on the depressed glycolysis induced by bradykinin in tissue damage.

Animals↗

Insulin or increased concentration of zinc in liver, kidney or muscle does not alter the phosphoglucomutase activity in vivo.

Injection of insulin into mice and rabbit has been reported to convert the latent form of phosphoglucomutase (E X Zn) to an active form (E X Mg) without any net increase in the protein associated with this enzyme (Hashimoto et al., Biochem. Biophys. Res. Commun., 27 (1967) 369 and Peck et al., J. Biol. Chem. 242 (1971) 1160). Because the injection of Cd in rats causes a 2-fold increase in Zn concentration in the liver, it appeared that such treatment in mice would increase the concentration of the latent enzyme and thereby enhance the insulin induced activation of the enzyme. The results however show that insulin or Cd treatment, alone or in combination do not alter the concentrations of the 2 forms of enzyme in liver, kidney or muscle of normal or fasted mice. In all instances the conversion of the latent enzyme to its activated form required preincubation with a chelating agent. Fractionation of the tissue homogenates showed that upon Cd injection the concentration of Zn in the fractions of Mr less and 1000 increased 6-fold in liver and 3-fold in kidney, that in muscle remained unchanged.

Animals↗

Human erythrocyte phosphoglucomutase: comparison of the kinetic properties of PGM1 and PGM2 isoenzymes.

Kinetic properties of PGM1 and PGM2 phosphoglucomutase "primary" isoenzymes from human erythrocytes were studied. The two enzyme forms share a "ping-pong" kinetic mechanism and show similar Km for substrate (glucose 1-P) and cofactor (glucose 1,6-P2). Micromolar concentrations of fructose 1,6-P2 and glycerate 2,3-P2 inhibit both PGM1 and PGM2 isoenzymes to a similar extent. The sole PGM2 form is affected by ribose monophosphates (ribose 1-P and ribose 5-P) that act as mutase inhibitors vs. glucose 1,6-P2 and as apparent activators vs. glucose 1-P. The interaction between PGM2 isoenzyme and ribose monophosphates is discussed in the light of the ability of this form to also display phosphoribomutase activity.

2,3-Diphosphoglycerate↗

Comparison of the properties of phosphoglucomutase from muscle of turbot (Reinhardtius hippoglossoides, Greenland Halibut) and witch flounder (Glyptocephalus cynoglossus) from the Northwest Atlantic and the Gulf of St. Lawrence.

1. Phosphoglucomutase from muscle of turbot and witch has been purified and the properties studied from two regions of the Northwest Atlantic and the Gulf of St. Lawrence. 2. The enzyme, purified 157 fold from turbot and 164 fold from witch had specific activities of 596 and 475 U/mg/min respectively. 3. The M.W. and pH optimum of the enzyme from both species were 63,500 and 7500; and 63,000 and 7800 for the Northwest Atlantic and the Gulf samples respectively. 4. The amino acid contents and responses towards different inhibitory agents were different for the enzyme from the Gulf than the Northwest Atlantic samples. 5. The findings compares well with the genetic analysis of the electrophoretic properties of the enzyme in relation to stock discrimination of the two species.

Amino Acids↗

Phosphoglucomutase polymorphism in the oriental fruit fly, Dacus dorsalis (Diptera, Tephritidae) from Peninsular Malaysia.

Seven natural populations of Dacus dorsalis were analysed for phosphoglucomutase by means of horizontal starch-gel electrophoresis. The electrophoretic phenotypes were governed by four codominant Pgm alleles. The commonest allele in all the seven population samples was PgmB which encoded an electrophoretic band with intermediate mobility. The distributions of PGM phenotype were in accordance with Hardy-Weinberg expectations. There was geographic variation in the distribution of Pgm alleles.

Alleles↗

Glucose-1,6-P2 synthesis, phosphoglucomutase and phosphoribomutase correlate with glucose-1,6-P2 concentration in mammals red blood cells.

Glucose 1,6-biphosphate (G1,6P2) was measured in human, pig, cow, rabbit, rat and sheep red blood cells. Mean values are variable among the species and range from 33 to 122 nmol/ml RBC for pig and rabbit erythrocytes, respectively. The activities of G1,6P2 synthase, phosphoglucomutase (PGM) and phosphoribomutase (PRM) have also been assayed in red cell haemolysates of the same species. The correlations between the biphosphate content and the occurrence of the three enzymatic activities have been studied in order to gain an insight into the regulation of the G1,6P2 turnover in mammalian erythrocytes.

Animals↗

Isolation and expression of a rat liver cDNA encoding phosphoglucomutase.

A cDNA encoding phosphoglucomutase (PGM) has been isolated from a rat liver cDNA library following screening with a polymerase chain reaction product. The cDNA was found to contain a 53-base-pair (bp) 5' untranslated region (5' UTR), a single start codon and consensus initiation sequence, an open reading frame (ORF) of 1686 bp, and a 3' untranslated tail. A comparison to the rabbit and human muscle PGM cDNAs [Whitehouse et al., Proc. Natl. Acad. Sci. USA 89 (1992) 411-415] showed 90% identity of rat cDNA to both, while a comparison to the deduced amino acid sequences showed 97 and 96% identity, respectively. Northern blot analyses determined that PGM was encoded by a single mRNA in rat liver, of approximately 2.2 kb. Following transfection of COS-7 cells with a plasmid containing the entire PGM ORF, indirect immunofluorescence analyses using a PGM-specific monoclonal antibody determined that approximately 5% of the cells displayed 50-100 times greater fluorescence than that seen in the remainder of the cells or in mock transfects. The enhanced production of PGM was also demonstrated by Western blotting and by enzymatic activity assays.

Amino Acid Sequence↗

A chromosomal cluster of genes encoding ADP-glucose synthetase, glycogen synthase and phosphoglucomutase in Agrobacterium tumefaciens.

A chromosomal region from Agrobacterium tumefaciens that complements exoC (pgm) mutations was cloned and sequenced. A cluster of three open reading frames (ORF1, ORF2 and ORF3) was identified. These genes are oriented in the same direction and are involved in the synthesis of glycogen and other polysaccharides. ORF1 encodes a 420-amino-acid (aa) protein with 55.9% homology to Escherichia coli GlgC (ADP-glucose synthetase, EC 2.7.7.27). ORF2 encodes a 480-aa protein with 42.2% homology to E. coli GlgA (glycogen synthase, EC 2.4.1.21). Based on Tn5 mutagenesis and protein homology, ORF3 was identified as the structural gene encoding phosphoglucomutase (Pgm, EC 2.7.5.1). ORF3 encodes a 542-aa protein with 52.6% homology to rabbit Pgm. There is no significant homology (less than 20%) to the Xanthomonas campestris XanA protein, which displays phosphomannomutase (Pmm) and Pgm activities [Koplin et al., J. Bacteriol 174 (1992) 191-199]. An A. tumefaciens pgm::Tn5 mutant retains Pmm activity.

Agrobacterium tumefaciens↗

Red cell PGM1 (phosphoglucomutase) phenotyping by isoelectric focussing and starch gel electrophoresis in cases of disputed paternity in the United Kingdom. An evaluation of the results obtained in 95 cases.

Using two techniques for red cell PGM1 (phosphoglucomutase) phenotyping, namely starch gel electrophoresis and isoelectric focussing, a parallel study has been carried out in 95 paternity cases. The results again confirm the genetic basis for the extended polymorphism revealed by isoelectric focussing. Fifteen men were excluded from paternity on the basis of isoelectric focussing compared with seven excluded by starch gel electrophoresis. This was found to be in good agreement with the predicted increase in exclusion rate that the isoelectric focussing technique for red cell PGM1 provides.

Electrophoresis, Starch Gel↗

Studies and observations on the use of isoelectric focusing in ultra-thin polyacrylamide gels as a method of typing human red cell phosphoglucomutase.

The technique of isoelectric focusing in ultra-thin polyacrylamide gels as a method of typing human red cell phosphoglucomutase (PGM1) has been studied. Typing was possible without the samples attaining true equilibrium focusing conditions. The isozyme patterns so obtained were clearly defined and free from distortion. The importance of assessing relative band intensities when interpreting the isozyme patterns is discussed. Our experience of using the technique to analyse casework material is described.

Acrylic Resins↗

Phosphoglucomutase isoenzymes in human teeth.

Powdered dentine samples were prepared from fresh teeth and teeth allowed to putrefy for up to 90 days. In some samples dentine only was included, whereas in others dentine plus pulp remnants were used. Extracts of the powders were tested for phosphoglucomutase activity by starch gel electrophoresis and PGM typing was carried out. Fresh teeth showed activity in at least 50% of cases as did putrefied dentine plus pulp remnants. Putrefied dentine alone showed minimal activity. The method may be useful for identification procedures using fragments of teeth.

Blood Grouping and Crossmatching↗

Population data on the forensic genetic markers: phosphoglucomutase-1, esterase D, erythrocyte acid phosphatase and glyoxylase I.

Blood specimens from white and black sample populations from Baltimore, Maryland, were analyzed for the four most forensically important, polymorphic red cell enzyme systems-phosphoglucomutase-1, esterase D, erythrocyte acid phosphatase and glyoxalase I. The distributions of the phenotypes for each marker in each racial group were in Hardy-Weinberg equilibrium. The population data were similar to previously reported data for Whites and Blacks from different geographical locations within the United States.

Acid Phosphatase↗

Red cell phosphoglucomutase (PGM1) subtypes in Egyptians.

The polymorphism of the human red cell phosphoglucomutase 1 (PGM1) in samples from Egyptians (n = 134) was investigated using isoelectric focusing in thin-layer polyacrylamide gel. In the studied population samples nine common phenotypes were observed, and the calculated frequencies for the genes PGM1+1, PGM1-1, PGM2+1 and PGM2-1 were 0.6381, 0.0821, 0.2201 and 0.0597, respectively. The observed and expected phenotypes provide a good fit to Hardy-Weinberg equilibrium. The four alleles system will increase the probability of excluding a man falsely accused of paternity to 30% as compared with 16% if the two alleles system is used.

Egypt↗

A comparison of the sensitivity of typing group-specific component (Gc) and the phosphoglucomutase (PGM1) locus in control and casework bloodstains by three isoelectric focusing methods.

The performance of typing group-specific component (Gc) in bloodstains by two isoelectric focusing methods followed by its detection with silver staining has been compared with an established forensic system of typing phosphoglucomutase (PGM1) locus phenotypes by isoelectric focusing (IEF) in 1 mm gels. For Gc typing ultra-thin isoelectric focusing (UTIEF) gels and immobilized pH gradient (IPG) gels were used. Both laboratory prepared stains and casework stains were examined. The Gc UTIEF method is approximately eight times more sensitive than the existing PGM1 1 mm IEF method for control and casework stains. However, on average, a larger amount of stain was taken from casework stains than control stains for each typing system. A total of 53 casework stains were examined. Comparable success rates of 62% and 64% were obtained for typing Gc on UTIEF gels and PGM1 by 1 mm IEF, respectively. A success rate of 55% was obtained for typing Gc on IPGs. Bloodstains that were over 200 days old were successfully grouped by all three methods.

Blood Stains↗

A comparison of the isoelectric focusing patterns of red cell phosphoglucomutase separated on an ampholine, ampholine/separator and immobilised pH gradient.

The isoelectric focusing patterns of red cell phosphoglucomutase (PGM1) separated on Ampholine, Ampholine/separator and rehydratable immobilised pH gradients have been compared. The sharpest and most intense zymograms have been observed on immobilised pH gradients provided that Ampholine was added during the rehydration of the gel. The addition of ampholine in the rehydration of Immobiline plates has been shown to improve the sharpness and intensity of the zymogram obtained and the potential of immobilised pH gradients for PGM typing has been demonstrated.

Ampholyte Mixtures↗

Phosphoglucomutase subtyping in a south Polish population.

Phosphoglucomutase (PGM1) subtypes in South Polish population were examined by thin-layer polyacrylamide gel isoelectrofocusing (pH 5-7) using fresh hemolysates from 460 unrelated adults. The allele frequencies in Polish population are as follows: PGM1+1 = 0.6402, PGM1-1 = 0.1185, PGM2+1 = 0.1880, PGM2-1 = 0.0533.

Adolescent↗

Immunological detection of human phosphoglucomutase (PGM 1) subtypes.

Anti-phosphoglucomutase (PGM) antibodies have been produced by immunising a sheep with a purified preparation of rabbit skeletal muscle PGM and used to devise an immunological procedure for detecting PGM isozymes after isoelectric focusing. The anti-rabbit PGM antibodies cross react with human PGM and can be used to identify the PGM1 isozymes characteristic of this polymorphism. The patterns revealed by immunodetection are exactly comparable with those obtained by isozyme staining.

Animals↗