Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1990 Feb;87(4):1476–1480. doi: 10.1073/pnas.87.4.1476

Anti-inositolglycan antibodies selectively block some of the actions of insulin in intact BC3H1 cells.

G Romero 1, G Gámez 1, L C Huang 1, K Lilley 1, L Luttrell 1
PMCID: PMC53498  PMID: 2137614

Abstract

We have studied the mechanism of generation of insulin mediators by using specific antibodies raised against the oligosaccharide anchor of membrane proteins. These antibodies (i) block the in vitro effects of purified insulin mediators and (ii) block the insulin-induced stimulation of pyruvate dehydrogenase in intact BC3H1 myocytes but not insulin-stimulated glucose uptake, generation of diacylglycerol, or generation of insulin mediators. When added to intact cells in the presence of insulin, these antibodies induce the accumulation of insulin mediator activity in the extracellular medium. We therefore conclude that these anti-inositolglycan antibodies block some of the effects of insulin by inhibiting the uptake of specific insulin mediators generated outside the cell.

Full text

PDF
1476

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alvarez J. F., Varela I., Ruiz-Albusac J. M., Mato J. M. Localisation of the insulin-sensitive phosphatidylinositol glycan at the outer surface of the cell membrane. Biochem Biophys Res Commun. 1988 May 16;152(3):1455–1462. doi: 10.1016/s0006-291x(88)80449-2. [DOI] [PubMed] [Google Scholar]
  2. Bangs J. D., Hereld D., Krakow J. L., Hart G. W., Englund P. T. Rapid processing of the carboxyl terminus of a trypanosome variant surface glycoprotein. Proc Natl Acad Sci U S A. 1985 May;82(10):3207–3211. doi: 10.1073/pnas.82.10.3207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Buss J. E., Stull J. T. Measurement of chemical phosphate in proteins. Methods Enzymol. 1983;99:7–14. doi: 10.1016/0076-6879(83)99035-3. [DOI] [PubMed] [Google Scholar]
  4. Cardoso de Almeida M. L., Turner M. J. The membrane form of variant surface glycoproteins of Trypanosoma brucei. Nature. 1983 Mar 24;302(5906):349–352. doi: 10.1038/302349a0. [DOI] [PubMed] [Google Scholar]
  5. Caro J. F., Folli F., Cecchin F., Sinha M. K. Chemical mediator of insulin action stimulates lipid synthesis and down regulates the insulin receptors in primary cultures of rat hepatocytes. Biochem Biophys Res Commun. 1983 Aug 30;115(1):375–382. doi: 10.1016/0006-291x(83)91014-8. [DOI] [PubMed] [Google Scholar]
  6. Chan B. L., Lisanti M. P., Rodriguez-Boulan E., Saltiel A. R. Insulin-stimulated release of lipoprotein lipase by metabolism of its phosphatidylinositol anchor. Science. 1988 Sep 23;241(4873):1670–1672. doi: 10.1126/science.241.4873.1670. [DOI] [PubMed] [Google Scholar]
  7. Farese R. V., Standaert M. L., Barnes D. E., Davis J. S., Pollet R. J. Phorbol ester provokes insulin-like effects on glucose transport, amino acid uptake, and pyruvate dehydrogenase activity in BC3H-1 cultured myocytes. Endocrinology. 1985 Jun;116(6):2650–2655. doi: 10.1210/endo-116-6-2650. [DOI] [PubMed] [Google Scholar]
  8. Ferguson M. A., Homans S. W., Dwek R. A., Rademacher T. W. Glycosyl-phosphatidylinositol moiety that anchors Trypanosoma brucei variant surface glycoprotein to the membrane. Science. 1988 Feb 12;239(4841 Pt 1):753–759. doi: 10.1126/science.3340856. [DOI] [PubMed] [Google Scholar]
  9. Ferguson M. A., Low M. G., Cross G. A. Glycosyl-sn-1,2-dimyristylphosphatidylinositol is covalently linked to Trypanosoma brucei variant surface glycoprotein. J Biol Chem. 1985 Nov 25;260(27):14547–14555. [PubMed] [Google Scholar]
  10. Hereld D., Krakow J. L., Bangs J. D., Hart G. W., Englund P. T. A phospholipase C from Trypanosoma brucei which selectively cleaves the glycolipid on the variant surface glycoprotein. J Biol Chem. 1986 Oct 15;261(29):13813–13819. [PubMed] [Google Scholar]
  11. Homans S. W., Ferguson M. A., Dwek R. A., Rademacher T. W., Anand R., Williams A. F. Complete structure of the glycosyl phosphatidylinositol membrane anchor of rat brain Thy-1 glycoprotein. Nature. 1988 May 19;333(6170):269–272. doi: 10.1038/333269a0. [DOI] [PubMed] [Google Scholar]
  12. Ikezawa H., Yamanegi M., Taguchi R., Miyashita T., Ohyabu T. Studies on phosphatidylinositol phosphodiesterase (phospholipase C type) of Bacillus cereus. I. purification, properties and phosphatase-releasing activity. Biochim Biophys Acta. 1976 Nov 19;450(2):154–164. [PubMed] [Google Scholar]
  13. Ishihara M., Fedarko N. S., Conrad H. E. Involvement of phosphatidylinositol and insulin in the coordinate regulation of proteoheparan sulfate metabolism and hepatocyte growth. J Biol Chem. 1987 Apr 5;262(10):4708–4716. [PubMed] [Google Scholar]
  14. Jarett L., Wong E. H., Macaulay S. L., Smith J. A. Insulin mediators from rat skeletal muscle have differential effects on insulin-sensitive pathways of intact adipocytes. Science. 1985 Feb 1;227(4686):533–535. doi: 10.1126/science.3917578. [DOI] [PubMed] [Google Scholar]
  15. Kennington A. S., Shen T. Y., Romero G. A simple procedure for the preparation and purification of the oligosaccharide components of the glycosylphosphatidylinositol anchor of membrane proteins. Anal Biochem. 1989 Aug 15;181(1):1–5. doi: 10.1016/0003-2697(89)90384-9. [DOI] [PubMed] [Google Scholar]
  16. Larner J., Huang L. C., Schwartz C. F., Oswald A. S., Shen T. Y., Kinter M., Tang G. Z., Zeller K. Rat liver insulin mediator which stimulates pyruvate dehydrogenase phosphate contains galactosamine and D-chiroinositol. Biochem Biophys Res Commun. 1988 Mar 30;151(3):1416–1426. doi: 10.1016/s0006-291x(88)80520-5. [DOI] [PubMed] [Google Scholar]
  17. Larner J., Huang L. C., Tang G., Suzuki S., Schwartz C. F., Romero G., Roulidis Z., Zeller K., Shen T. Y., Oswald A. S. Insulin mediators: structure and formation. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 2):965–971. doi: 10.1101/sqb.1988.053.01.111. [DOI] [PubMed] [Google Scholar]
  18. Low M. G., Finean J. B. Non-lytic release of acetylcholinesterase from erythrocytes by a phosphatidylinositol-specific phospholipase C. FEBS Lett. 1977 Oct 1;82(1):143–146. doi: 10.1016/0014-5793(77)80905-8. [DOI] [PubMed] [Google Scholar]
  19. Low M. G., Finean J. B. Specific release of plasma membrane enzymes by a phosphatidylinositol-specific phospholipase C. Biochim Biophys Acta. 1978 Apr 20;508(3):565–570. doi: 10.1016/0005-2736(78)90100-1. [DOI] [PubMed] [Google Scholar]
  20. Low M. G., Kincade P. W. Phosphatidylinositol is the membrane-anchoring domain of the Thy-1 glycoprotein. Nature. 1985 Nov 7;318(6041):62–64. doi: 10.1038/318062a0. [DOI] [PubMed] [Google Scholar]
  21. Luttrell L., Rogol A. D. Partial insulin resistance in the mouse BC3H-1 cell line: absent hexose-independent actions of insulin. Endocrinology. 1986 Jul;119(1):331–342. doi: 10.1210/endo-119-1-331. [DOI] [PubMed] [Google Scholar]
  22. Malchoff C. D., Huang L., Gillespie N., Palasi C. V., Schwartz C. F., Cheng K., Hewlett E. L., Larner J. A putative mediator of insulin action which inhibits adenylate cyclase and adenosine 3',5'-monophosphate-dependent protein kinase: partial purification from rat liver: site and kinetic mechanism of action. Endocrinology. 1987 Apr;120(4):1327–1337. doi: 10.1210/endo-120-4-1327. [DOI] [PubMed] [Google Scholar]
  23. Mato J. M., Kelly K. L., Abler A., Jarett L. Identification of a novel insulin-sensitive glycophospholipid from H35 hepatoma cells. J Biol Chem. 1987 Feb 15;262(5):2131–2137. [PubMed] [Google Scholar]
  24. Muchmore D. B., Raess B. U., Bergstrom R. W., de Haën C. On the mechanisms of inhibition of insulin action by small-molecular-weight trypsin inhibitors. Diabetes. 1982 Nov;31(11):976–984. doi: 10.2337/diacare.31.11.976. [DOI] [PubMed] [Google Scholar]
  25. Newman J. D., Armstrong J. M., Bornstein J. Assay of insulin mediator activity with soluble pyruvate dehydrogenase phosphatase. Endocrinology. 1985 May;116(5):1912–1919. doi: 10.1210/endo-116-5-1912. [DOI] [PubMed] [Google Scholar]
  26. Rinaudo M. T., Curto M., Bruno R., Marino C., Rossetti V., Mostert M. Evidence of an insulin generated pyruvate dehydrogenase stimulating factor in rat brain plasma membranes. Int J Biochem. 1987;19(10):909–913. doi: 10.1016/0020-711x(87)90171-6. [DOI] [PubMed] [Google Scholar]
  27. Roberts W. L., Santikarn S., Reinhold V. N., Rosenberry T. L. Structural characterization of the glycoinositol phospholipid membrane anchor of human erythrocyte acetylcholinesterase by fast atom bombardment mass spectrometry. J Biol Chem. 1988 Dec 15;263(35):18776–18784. [PubMed] [Google Scholar]
  28. Romero G., Luttrell L., Rogol A., Zeller K., Hewlett E., Larner J. Phosphatidylinositol-glycan anchors of membrane proteins: potential precursors of insulin mediators. Science. 1988 Apr 22;240(4851):509–511. doi: 10.1126/science.3282305. [DOI] [PubMed] [Google Scholar]
  29. Saltiel A. R., Cuatrecasas P. Insulin stimulates the generation from hepatic plasma membranes of modulators derived from an inositol glycolipid. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5793–5797. doi: 10.1073/pnas.83.16.5793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Saltiel A. R., Fox J. A., Sherline P., Cuatrecasas P. Insulin-stimulated hydrolysis of a novel glycolipid generates modulators of cAMP phosphodiesterase. Science. 1986 Aug 29;233(4767):967–972. doi: 10.1126/science.3016898. [DOI] [PubMed] [Google Scholar]
  31. Saltiel A. R., Sorbara-Cazan L. R. Inositol glycan mimics the action of insulin on glucose utilization in rat adipocytes. Biochem Biophys Res Commun. 1987 Dec 31;149(3):1084–1092. doi: 10.1016/0006-291x(87)90519-5. [DOI] [PubMed] [Google Scholar]
  32. Sato T., Palasi C. V., Huang L., Tang G., Larner A. C., Larner J. Insulin and a putative insulin metabolic mediator fraction from liver and muscle stimulate p33 messenger ribonucleic acid accumulation by apparently different mechanisms. Endocrinology. 1988 Sep;123(3):1559–1564. doi: 10.1210/endo-123-3-1559. [DOI] [PubMed] [Google Scholar]
  33. Standaert M. L., Farese R. V., Cooper D. R., Pollet R. J. Insulin-induced glycerolipid mediators and the stimulation of glucose transport in BC3H-1 myocytes. J Biol Chem. 1988 Jun 25;263(18):8696–8705. [PubMed] [Google Scholar]
  34. Strålfors P. Insulin stimulation of glucose uptake can be mediated by diacylglycerol in adipocytes. Nature. 1988 Oct 6;335(6190):554–556. doi: 10.1038/335554a0. [DOI] [PubMed] [Google Scholar]
  35. Whatmore A. J., Spitalnik S. L., Gaulton G. N., Jarett L. Initial characterization of a polyclonal antibody to an insulin-sensitive glycophospholipid. Arch Biochem Biophys. 1988 Jul;264(1):355–360. doi: 10.1016/0003-9861(88)90606-6. [DOI] [PubMed] [Google Scholar]
  36. Witters L. A., Watts T. D. An autocrine factor from Reuber hepatoma cells that stimulates DNA synthesis and acetyl-CoA carboxylase. Characterization of biologic activity and evidence for a glycan structure. J Biol Chem. 1988 Jun 15;263(17):8027–8036. [PubMed] [Google Scholar]
  37. Zhang S. R., Shi Q. H., Ho R. J. Cyclic AMP-lowering mediator of insulin. J Biol Chem. 1983 May 25;258(10):6471–6476. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES