Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1998 Oct 15;335(Pt 2):351–356. doi: 10.1042/bj3350351

Phosphatidylinositol 4-kinase, but not phosphatidylinositol 3-kinase, is present in GLUT4-containing vesicles isolated from rat skeletal muscle.

S Kristiansen 1, T Ramlal 1, A Klip 1
PMCID: PMC1219789  PMID: 9761734

Abstract

Insulin stimulates the rate of glucose transport into muscle and adipose cells by translocation of glucose transporter (GLUT4)-containing vesicles from an intracellular storage pool to the surface membrane. This event is mediated through the insulin receptor substrates (IRSs), which in turn activate phosphatidylinositol (PI) 3-kinase isoforms. It has been suggested that insulin causes attachment of PI 3-kinases to the intracellular GLUT4-containing vesicles in rat adipose cells. Furthermore, it has also been shown that GLUT4-containing vesicles in adipose cells contain a PI 4-kinase. In the present study we investigate whether GLUT4-containing vesicles isolated from rat skeletal muscle display PI 3-kinase and/or PI 4-kinase activities. Insulin stimulation caused a rapid increase (5-15-fold increase compared with control) in the intracellular cytosolic IRS-1-associated PI-3 kinase activity. This PI 3-kinase activity was also present in a membrane preparation containing the insulin-regulatable pool of GLUT4 transporters. However, when GLUT4-containing vesicles were isolated by immunoprecipitation from basal and insulin-stimulated (3 min) skeletal muscle, the vesicles displayed PI 4-kinase, but not PI 3-kinase, activity. Insulin did not regulate the PI 4-kinase activity in the GLUT4-containing vesicles. In conclusion, GLUT4-containing vesicles from rat skeletal muscle contain a PI 4-kinase, but not a PI 3-kinase. It is suggested that, in skeletal muscle, insulin causes activation of the IRS/PI 3-kinase complex in an intracellular membrane compartment associated closely with the GLUT4-containing vesicles, but not in the GLUT4-containing vesicles themselves.

Full Text

The Full Text of this article is available as a PDF (473.2 KB).

Selected References

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

  1. Biber J. W., Lienhard G. E. Isolation of vesicles containing insulin-responsive, intracellular glucose transporters from 3T3-L1 adipocytes. J Biol Chem. 1986 Dec 5;261(34):16180–16184. [PubMed] [Google Scholar]
  2. Clark S. F., Martin S., Carozzi A. J., Hill M. M., James D. E. Intracellular localization of phosphatidylinositide 3-kinase and insulin receptor substrate-1 in adipocytes: potential involvement of a membrane skeleton. J Cell Biol. 1998 Mar 9;140(5):1211–1225. doi: 10.1083/jcb.140.5.1211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Del Vecchio R. L., Pilch P. F. Phosphatidylinositol 4-kinase is a component of glucose transporter (GLUT 4)-containing vesicles. J Biol Chem. 1991 Jul 15;266(20):13278–13283. [PubMed] [Google Scholar]
  4. Elmendorf J. S., Damrau-Abney A., Smith T. R., David T. S., Turinsky J. Insulin-stimulated phosphatidylinositol 3-kinase activity and 2-deoxy-D-glucose uptake in rat skeletal muscles. Biochem Biophys Res Commun. 1995 Mar 28;208(3):1147–1153. doi: 10.1006/bbrc.1995.1453. [DOI] [PubMed] [Google Scholar]
  5. Folli F., Saad M. J., Backer J. M., Kahn C. R. Regulation of phosphatidylinositol 3-kinase activity in liver and muscle of animal models of insulin-resistant and insulin-deficient diabetes mellitus. J Clin Invest. 1993 Oct;92(4):1787–1794. doi: 10.1172/JCI116768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hanpeter D., James D. E. Characterization of the intracellular GLUT-4 compartment. Mol Membr Biol. 1995 Jul-Sep;12(3):263–269. doi: 10.3109/09687689509072426. [DOI] [PubMed] [Google Scholar]
  7. Heller-Harrison R. A., Morin M., Guilherme A., Czech M. P. Insulin-mediated targeting of phosphatidylinositol 3-kinase to GLUT4-containing vesicles. J Biol Chem. 1996 Apr 26;271(17):10200–10204. doi: 10.1074/jbc.271.17.10200. [DOI] [PubMed] [Google Scholar]
  8. Kandror K. V., Coderre L., Pushkin A. V., Pilch P. F. Comparison of glucose-transporter-containing vesicles from rat fat and muscle tissues: evidence for a unique endosomal compartment. Biochem J. 1995 Apr 15;307(Pt 2):383–390. doi: 10.1042/bj3070383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Katagiri H., Asano T., Inukai K., Ogihara T., Ishihara H., Shibasaki Y., Murata T., Terasaki J., Kikuchi M., Yazaki Y. Roles of PI 3-kinase and Ras on insulin-stimulated glucose transport in 3T3-L1 adipocytes. Am J Physiol. 1997 Feb;272(2 Pt 1):E326–E331. doi: 10.1152/ajpendo.1997.272.2.E326. [DOI] [PubMed] [Google Scholar]
  10. Kelly K. L., Ruderman N. B. Insulin-stimulated phosphatidylinositol 3-kinase. Association with a 185-kDa tyrosine-phosphorylated protein (IRS-1) and localization in a low density membrane vesicle. J Biol Chem. 1993 Feb 25;268(6):4391–4398. [PubMed] [Google Scholar]
  11. Klip A., Ramlal T., Young D. A., Holloszy J. O. Insulin-induced translocation of glucose transporters in rat hindlimb muscles. FEBS Lett. 1987 Nov 16;224(1):224–230. doi: 10.1016/0014-5793(87)80452-0. [DOI] [PubMed] [Google Scholar]
  12. Kotani K., Carozzi A. J., Sakaue H., Hara K., Robinson L. J., Clark S. F., Yonezawa K., James D. E., Kasuga M. Requirement for phosphoinositide 3-kinase in insulin-stimulated GLUT4 translocation in 3T3-L1 adipocytes. Biochem Biophys Res Commun. 1995 Apr 6;209(1):343–348. doi: 10.1006/bbrc.1995.1509. [DOI] [PubMed] [Google Scholar]
  13. Lavoie L., He L., Ramlal T., Ackerley C., Marette A., Klip A. The GLUT4 glucose transporter and the alpha 2 subunit of the Na+,K(+)-ATPase do not localize to the same intracellular vesicles in rat skeletal muscle. FEBS Lett. 1995 Jun 12;366(2-3):109–114. doi: 10.1016/0014-5793(95)00507-6. [DOI] [PubMed] [Google Scholar]
  14. Lee A. D., Hansen P. A., Holloszy J. O. Wortmannin inhibits insulin-stimulated but not contraction-stimulated glucose transport activity in skeletal muscle. FEBS Lett. 1995 Mar 13;361(1):51–54. doi: 10.1016/0014-5793(95)00147-2. [DOI] [PubMed] [Google Scholar]
  15. Meyers R., Cantley L. C. Cloning and characterization of a wortmannin-sensitive human phosphatidylinositol 4-kinase. J Biol Chem. 1997 Feb 14;272(7):4384–4390. doi: 10.1074/jbc.272.7.4384. [DOI] [PubMed] [Google Scholar]
  16. Nakanishi S., Catt K. J., Balla T. A wortmannin-sensitive phosphatidylinositol 4-kinase that regulates hormone-sensitive pools of inositolphospholipids. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5317–5321. doi: 10.1073/pnas.92.12.5317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ogihara T., Shin B. C., Anai M., Katagiri H., Inukai K., Funaki M., Fukushima Y., Ishihara H., Takata K., Kikuchi M. Insulin receptor substrate (IRS)-2 is dephosphorylated more rapidly than IRS-1 via its association with phosphatidylinositol 3-kinase in skeletal muscle cells. J Biol Chem. 1997 May 9;272(19):12868–12873. doi: 10.1074/jbc.272.19.12868. [DOI] [PubMed] [Google Scholar]
  18. Okada T., Hazeki O., Ui M., Katada T. Synergistic activation of PtdIns 3-kinase by tyrosine-phosphorylated peptide and beta gamma-subunits of GTP-binding proteins. Biochem J. 1996 Jul 15;317(Pt 2):475–480. doi: 10.1042/bj3170475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pike L. J. Phosphatidylinositol 4-kinases and the role of polyphosphoinositides in cellular regulation. Endocr Rev. 1992 Nov;13(4):692–706. doi: 10.1210/edrv-13-4-692. [DOI] [PubMed] [Google Scholar]
  20. Quon M. J., Butte A. J., Zarnowski M. J., Sesti G., Cushman S. W., Taylor S. I. Insulin receptor substrate 1 mediates the stimulatory effect of insulin on GLUT4 translocation in transfected rat adipose cells. J Biol Chem. 1994 Nov 11;269(45):27920–27924. [PubMed] [Google Scholar]
  21. Rodnick K. J., Slot J. W., Studelska D. R., Hanpeter D. E., Robinson L. J., Geuze H. J., James D. E. Immunocytochemical and biochemical studies of GLUT4 in rat skeletal muscle. J Biol Chem. 1992 Mar 25;267(9):6278–6285. [PubMed] [Google Scholar]
  22. Saltiel A. R. Diverse signaling pathways in the cellular actions of insulin. Am J Physiol. 1996 Mar;270(3 Pt 1):E375–E385. doi: 10.1152/ajpendo.1996.270.3.E375. [DOI] [PubMed] [Google Scholar]
  23. Tanti J. F., Grémeaux T., Grillo S., Calleja V., Klippel A., Williams L. T., Van Obberghen E., Le Marchand-Brustel Y. Overexpression of a constitutively active form of phosphatidylinositol 3-kinase is sufficient to promote Glut 4 translocation in adipocytes. J Biol Chem. 1996 Oct 11;271(41):25227–25232. doi: 10.1074/jbc.271.41.25227. [DOI] [PubMed] [Google Scholar]
  24. Walsh J. P., Caldwell K. K., Majerus P. W. Formation of phosphatidylinositol 3-phosphate by isomerization from phosphatidylinositol 4-phosphate. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):9184–9187. doi: 10.1073/pnas.88.20.9184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang Q., Bilan P. J., Tsakiridis T., Hinek A., Klip A. Actin filaments participate in the relocalization of phosphatidylinositol3-kinase to glucose transporter-containing compartments and in the stimulation of glucose uptake in 3T3-L1 adipocytes. Biochem J. 1998 May 1;331(Pt 3):917–928. doi: 10.1042/bj3310917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Yeh J. I., Gulve E. A., Rameh L., Birnbaum M. J. The effects of wortmannin on rat skeletal muscle. Dissociation of signaling pathways for insulin- and contraction-activated hexose transport. J Biol Chem. 1995 Feb 3;270(5):2107–2111. doi: 10.1074/jbc.270.5.2107. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES