Abstract
The anomeric specificity of d-glucose metabolism in intact hepatocytes remains a matter of debate. This issue was further investigated in the present study, which is based on the quantification of the α- and β-anomers of the 13C-enriched isotopomers of d-glucose generated by rat liver cells exposed to either d-[1-13C] fructose or d-[2-13C] fructose in the presence of D2O. The d-[1-13C]glucose/d-[6-13C]glucose paired ratios found in the cells exposed to d-[1-13C] fructose and the d-[2-13C]glucose/d-[5-13C]glucose paired ratios found in the cells exposed to d-[2-13C] fructose yielded a paired β/α ratio averaging (mean ± S.E.M.) 79.3 ± 6.1%. In the case of the isotopomers of d-glucose formed by gluconeogenesis, the d-[2-13C]glucose/d-[5-13C]glucose and d-[3-13C]glucose/d-[4-13C]glucose paired ratios found in cells exposed to d-[1-13C] fructose, as well as the d-[1-13C]glucose/d-[6-13C]glucose and d-[3-13C]glucose/d-[4-13C]glucose paired ratios found in cells exposed to d-[2-13C]fructose, yielded an α/β paired ratio averaging 75.0 ± 5.8%. Last, in the cells exposed to d-[2-13C]fructose, the β/α ratio for the C2-deuterated isotopomers of d-[2-13C]glucose represented 78.9 ± 3.7% of that for the C5-deuterated isotopomers of d-[5-13C]glucose. The three values representative of the anomeric specificity of d-glucose production by liver cells were not significantly different from one another, with an overall mean value of 76.9 ± 3.6%. These findings unambiguously document that the anomeric specificity of phosphoglucoisomerase is operative in intact hepatocytes, resulting in a preferential output of the α-anomer of 13C-enriched d-glucose under the present experimental conditions (Mol Cell Biochem 266: 145–150, 2004)
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References
Hill JB: A method for measuring deviations from equilibrium of the glucose anomers in blood. J Appl Physiol 20: 749–754, 1965
Miwa I, Fujii H, Okuda J: Glycogen synthesis from glucose anomers in rat diaphragm. Abstracts of the 12th International Congress of Biochemistry, Perth, p 123, 1982
Malaisse WJ: Anomeric specificity of D-glucose utilization in rat hepatocytes. IRCS Med Sci 14: 609–610, 1986
Zähner D, Rasschaert J, Malaisse WJ: Anomeric specificity of liver glycogenolysis. Biochim Biophys Acta 926: 115–118, 1987
Bollen M, Malaisse-Lagae F, Malaisse W, Stalmans W: The interaction of phosphorylase a with D-glucose displays α-stereospecificity. Biochim Biophys Acta 1038: 141–145, 1990
Seglen PO: Preparation of isolated rat liver cells. Methods Cell Biol 13: 29–83, 1976
Malaisse WJ, Maggetto C, Leclercq-Meyer V, Sener A: Interference of glycogenolysis with glycolysis in pancreatic islets from glucose-infused rats. J Clin Invest 91: 432–436, 1993
Willem R, Biesemans M, Kayser F, Malaisse WJ: 13C NMR study of the generation of C2-and C3-deuterated lactic acid by tumoral pancreatic islet cells exposed to D-[1-13C]-, D-[2-13C]-and D-[6-13C]-glucose in 2H2O. Magn Reson Med 31: 259–267, 1994
Laatikainen R, Niemitz M, Malaisse WJ, Biesemans M, Willem R: Acomputational strategy for the deconvolution of NMR spectra with multiplet structures: Analysis of overlapping 13C—2H multiplets of 13C enriched metabolites from cell suspensions incubated in deuterated media. Magn Reson Med 36: 359–365, 1996
Leclercq-Meyer V, Malaisse-Lagae F, Coulic V, Akkan AG, Malaisse WJ: Preservation of the anomeric specificity of glucose-induced insulin release in partially pancreatectomized rats. Diabetologia 35: 505–509, 1992
Malaisse WJ, Ladrière L, Verbruggen I, Willem R: Metabolism of D-[1-13C]fructose, D-[2-13C]fructose, and D-[6-13C]fructose in rat hepa-tocytes incubated in the presence of H2O or D2O. Mol Gen Metab 75: 162–167, 2002
Malaisse WJ, Ladrière L, Verbruggen I, Willem R: Effects of D-glucose upon D-fructose metabolism in rat hepatocytes: A 13C-NMR study. Mol Cell Biochem 241: 103–106, 2002
Malaisse WJ, Willem R: Metabolism of 13C-enriched D-fructose in hepatocytes from Goto-Kakizaki rats. Abstracts of the 185th Meeting of the Belgian Society of Biochemistry, Namur, abstract 22, 2003
Willem R, Biesemans M, Hallenga K, Lippens G, Malaisse-Lagae F, Malaisse WJ: Dual anomeric specificity and dual anomerase activity of phosphoglucoisomerase quantified by two-dimensional phase sensitive 13C EXSY NMR. J Biol Chem 267: 210–217, 1992
Malaisse WJ, Zhang T-M, Verbruggen I, Willem R: D-glucose generation from [2-13C]pyruvate in rat hepatocytes: Implications in terms of enzyme-to-enzyme channelling. Arch Biochem Biophys 332: 341–351, 1996
Verbruggen I, Ladrière L, Willem R, Malaisse WJ: Asymmetrical labelling of D-glucose generated from [3-13C]pyruvate in rat hepatocytes. Biochem Mol Med 61: 229–235, 1997
Malaisse WJ, Malaisse-Lagae F, Liemans V, Ottinger R, Willem R: Phosphoglucoisomerase-catalyzed interconversion of hexose phosphates: Isotopic discrimination between hydrogen and deuterium. Mol Cell Biochem 93: 153–165, 1990
Furuya E, Hott K, Tagawa K: Anomeric specificity of glucose-6-phosphatase and glucokinase. Biochem Biophys Res Comm 141: 931–936, 1986
Ramrez R, Zähner D, Marynissen G, Sener A, Malaisse WJ: Anomeric specificity of D-glucose phosphorylation by rat liver glucose-6-phosphatase. Biochem J 261: 509–513, 1989
Malaisse WJ, Zhang Y, Jijakli H, Courtois P, Sener A: Enzyme-to-enzyme channelling of early glycolytic metabolites in rat pancreatic islets. Eur J Biochem 270(suppl. 1): 59 (abstract), 2003
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Malaisse, W., Willem, R. Anomeric specificity of D-glucose production by rat hepatocytes. Mol Cell Biochem 266, 145–150 (2004). https://doi.org/10.1023/B:MCBI.0000049152.06259.4b
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DOI: https://doi.org/10.1023/B:MCBI.0000049152.06259.4b