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. 1997 Jun 15;324(Pt 3):981–985. doi: 10.1042/bj3240981

Increased flux through the hexosamine biosynthesis pathway inhibits glucose transport acutely by activation of protein kinase C.

A Filippis 1, S Clark 1, J Proietto 1
PMCID: PMC1218517  PMID: 9210425

Abstract

The hexosamine biosynthesis pathway and protein kinase C (PKC) activation mediate hyperglycaemia-induced impaired glucose transport, but the relative role of each pathway is unknown. Following a 2 h preincubation of rat adipocytes in the presence of either high glucose (30 mM) plus insulin (0.7 nM) or glucosamine (3 mM), both high glucose and glucosamine inhibited subsequent basal and insulin-stimulated glucose transport, measured at 5.0 mM glucose. Azaserine, an inhibitor of the enzyme glutamine:fructose-6-phosphate aminotransferase, abolished the effect of high glucose, but not that of glucosamine. Ro-31-8220, an inhibitor of PKC, reversed the effects of both high glucose and glucosamine, suggesting that flux through the hexosamine biosynthesis pathway impaired glucose transport acutely by activating PKC. Both high glucose and glucosamine caused a 3-fold increase in PKC activity; this effect of high glucose, but not that of glucosamine, was partially decreased by azaserine. Neither high glucose nor glucosamine altered basal or insulin-stimulated plasma membrane GLUT1 levels, whereas both treatments decreased basal, but not insulin-stimulated, GLUT4 levels. Azaserine abolished the effect of high glucose, but not that of glucosamine, on basal plasma membrane GLUT4 levels. Ro-31-8220, which returned glucose transport to control values, caused a further decrease in plasma membrane GLUT4 levels. It is concluded that, in rat adipocytes, an acute increase in flux through the hexosamine biosynthesis pathway inhibits glucose transport by activation of PKC.

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Selected References

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  1. Allard W. J., Gibbs E. M., Witters L. A., Lienhard G. E. The glucose transporter in human fibroblasts is phosphorylated in response to phorbol ester but not in response to growth factors. Biochim Biophys Acta. 1987 Jul 29;929(3):288–295. doi: 10.1016/0167-4889(87)90255-2. [DOI] [PubMed] [Google Scholar]
  2. Baron A. D., Zhu J. S., Zhu J. H., Weldon H., Maianu L., Garvey W. T. Glucosamine induces insulin resistance in vivo by affecting GLUT 4 translocation in skeletal muscle. Implications for glucose toxicity. J Clin Invest. 1995 Dec;96(6):2792–2801. doi: 10.1172/JCI118349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bollag G. E., Roth R. A., Beaudoin J., Mochly-Rosen D., Koshland D. E., Jr Protein kinase C directly phosphorylates the insulin receptor in vitro and reduces its protein-tyrosine kinase activity. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5822–5824. doi: 10.1073/pnas.83.16.5822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  6. Chin J. E., Liu F., Roth R. A. Activation of protein kinase C alpha inhibits insulin-stimulated tyrosine phosphorylation of insulin receptor substrate-1. Mol Endocrinol. 1994 Jan;8(1):51–58. doi: 10.1210/mend.8.1.7512195. [DOI] [PubMed] [Google Scholar]
  7. Craven P. A., DeRubertis F. R. Protein kinase C is activated in glomeruli from streptozotocin diabetic rats. Possible mediation by glucose. J Clin Invest. 1989 May;83(5):1667–1675. doi: 10.1172/JCI114066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Draznin B., Leitner J. W., Sussman K. E., Sherman N. A. Insulin and glucose modulate protein kinase C activity in rat adipocytes. Biochem Biophys Res Commun. 1988 Oct 14;156(1):570–575. doi: 10.1016/s0006-291x(88)80880-5. [DOI] [PubMed] [Google Scholar]
  9. Dunlop M. E., Larkins R. G. Pancreatic islets synthesize phospholipids de novo from glucose via acyl-dihydroxyacetone phosphate. Biochem Biophys Res Commun. 1985 Oct 30;132(2):467–473. doi: 10.1016/0006-291x(85)91157-x. [DOI] [PubMed] [Google Scholar]
  10. Dunlop M. E., Larkins R. G. Pancreatic islets synthesize phospholipids de novo from glucose via acyl-dihydroxyacetone phosphate. Biochem Biophys Res Commun. 1985 Oct 30;132(2):467–473. doi: 10.1016/0006-291x(85)91157-x. [DOI] [PubMed] [Google Scholar]
  11. Farese R. V., Standaert M. L., Arnold T. P., Yamada K., Musunuru K., Hernandez H., Mischak H., Cooper D. R. Preferential activation of microsomal diacylglycerol/protein kinase C signaling during glucose treatment (De Novo phospholipid synthesis) of rat adipocytes. J Clin Invest. 1994 May;93(5):1894–1899. doi: 10.1172/JCI117180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fisher M. D., Frost S. C. Translocation of GLUT1 does not account for elevated glucose transport in glucose-deprived 3T3-L1 adipocytes. J Biol Chem. 1996 May 17;271(20):11806–11809. doi: 10.1074/jbc.271.20.11806. [DOI] [PubMed] [Google Scholar]
  13. Galante P., Mosthaf L., Kellerer M., Berti L., Tippmer S., Bossenmaier B., Fujiwara T., Okuno A., Horikoshi H., Häring H. U. Acute hyperglycemia provides an insulin-independent inducer for GLUT4 translocation in C2C12 myotubes and rat skeletal muscle. Diabetes. 1995 Jun;44(6):646–651. doi: 10.2337/diab.44.6.646. [DOI] [PubMed] [Google Scholar]
  14. Garvey W. T., Olefsky J. M., Matthaei S., Marshall S. Glucose and insulin co-regulate the glucose transport system in primary cultured adipocytes. A new mechanism of insulin resistance. J Biol Chem. 1987 Jan 5;262(1):189–197. [PubMed] [Google Scholar]
  15. Häring H. U., Biermann E., Kemmler W. Coupling of insulin binding and insulin action on glucose transport in fat cells. Am J Physiol. 1981 May;240(5):E556–E565. doi: 10.1152/ajpendo.1981.240.5.E556. [DOI] [PubMed] [Google Scholar]
  16. Kelada A. S., Macaulay S. L., Proietto J. Cyclic AMP acutely stimulates translocation of the major insulin-regulatable glucose transporter GLUT4. J Biol Chem. 1992 Apr 5;267(10):7021–7025. [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Lee T. S., MacGregor L. C., Fluharty S. J., King G. L. Differential regulation of protein kinase C and (Na,K)-adenosine triphosphatase activities by elevated glucose levels in retinal capillary endothelial cells. J Clin Invest. 1989 Jan;83(1):90–94. doi: 10.1172/JCI113889. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  19. Marshall S., Bacote V., Traxinger R. R. Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system. Role of hexosamine biosynthesis in the induction of insulin resistance. J Biol Chem. 1991 Mar 15;266(8):4706–4712. [PubMed] [Google Scholar]
  20. Müller H. K., Kellerer M., Ermel B., Mühlhöfer A., Obermaier-Kusser B., Vogt B., Häring H. U. Prevention by protein kinase C inhibitors of glucose-induced insulin-receptor tyrosine kinase resistance in rat fat cells. Diabetes. 1991 Nov;40(11):1440–1448. doi: 10.2337/diab.40.11.1440. [DOI] [PubMed] [Google Scholar]
  21. Napoli R., Hirshman M. F., Horton E. S. Mechanisms and time course of impaired skeletal muscle glucose transport activity in streptozocin diabetic rats. J Clin Invest. 1995 Jul;96(1):427–437. doi: 10.1172/JCI118053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Olefsky J. M. Mechanisms of the ability of insulin to activate the glucose-transport system in rat adipocytes. Biochem J. 1978 Apr 15;172(1):137–145. doi: 10.1042/bj1720137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. RODBELL M. METABOLISM OF ISOLATED FAT CELLS. I. EFFECTS OF HORMONES ON GLUCOSE METABOLISM AND LIPOLYSIS. J Biol Chem. 1964 Feb;239:375–380. [PubMed] [Google Scholar]
  24. Robinson K. A., Sens D. A., Buse M. G. Pre-exposure to glucosamine induces insulin resistance of glucose transport and glycogen synthesis in isolated rat skeletal muscles. Study of mechanisms in muscle and in rat-1 fibroblasts overexpressing the human insulin receptor. Diabetes. 1993 Sep;42(9):1333–1346. doi: 10.2337/diab.42.9.1333. [DOI] [PubMed] [Google Scholar]
  25. Robinson K. A., Weinstein M. L., Lindenmayer G. E., Buse M. G. Effects of diabetes and hyperglycemia on the hexosamine synthesis pathway in rat muscle and liver. Diabetes. 1995 Dec;44(12):1438–1446. doi: 10.2337/diab.44.12.1438. [DOI] [PubMed] [Google Scholar]
  26. Rossetti L., Smith D., Shulman G. I., Papachristou D., DeFronzo R. A. Correction of hyperglycemia with phlorizin normalizes tissue sensitivity to insulin in diabetic rats. J Clin Invest. 1987 May;79(5):1510–1515. doi: 10.1172/JCI112981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Takayama S., White M. F., Kahn C. R. Phorbol ester-induced serine phosphorylation of the insulin receptor decreases its tyrosine kinase activity. J Biol Chem. 1988 Mar 5;263(7):3440–3447. [PubMed] [Google Scholar]
  28. Thomas T. P., Gopalakrishna R., Anderson W. B. Hormone- and tumor promoter-induced activation or membrane association of protein kinase C in intact cells. Methods Enzymol. 1987;141:399–411. doi: 10.1016/0076-6879(87)41086-0. [DOI] [PubMed] [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Traxinger R. R., Marshall S. Role of amino acids in modulating glucose-induced desensitization of the glucose transport system. J Biol Chem. 1989 Dec 15;264(35):20910–20916. [PubMed] [Google Scholar]
  31. Traxinger R. R., Marshall S. Suitability of 2-deoxyglucose for measuring initial rates of glucose uptake in isolated adipocytes. Biochem Int. 1990 Nov;22(4):607–615. [PubMed] [Google Scholar]
  32. Vogt B., Mushack J., Seffer E., Häring H. U. The translocation of the glucose transporter sub-types GLUT1 and GLUT4 in isolated fat cells is differently regulated by phorbol esters. Biochem J. 1991 May 1;275(Pt 3):597–600. doi: 10.1042/bj2750597. [DOI] [PMC free article] [PubMed] [Google Scholar]

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