Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Oct 1;367(Pt 1):301–306. doi: 10.1042/BJ20020708

Regulation of insulin signal transduction pathway by a small-molecule insulin receptor activator.

Victor D H Ding 1, Sajjad A Qureshi 1, Deborah Szalkowski 1, Zhihua Li 1, Dawn E Biazzo-Ashnault 1, Dan Xie 1, Kun Liu 1, A Brian Jones 1, David E Moller 1, Bei B Zhang 1
PMCID: PMC1222849  PMID: 12036431

Abstract

Insulin regulates cellular metabolism and growth through activation of insulin receptors (IRs). We recently identified a non-peptide small-molecule IR activator (compound 2), which induced human IR tyrosine kinase activity in Chinese-hamster ovary cells expressing human IR [Qureshi, Ding, Li, Szalkowski, Biazzo-Ashnault, Xie, Saperstein, Brady, Huskey, Shen et al. (2000) J. Biol. Chem. 275, 36590-36595]. Oral treatment with this compound resulted in correction of hyperglycaemia, hypertriacylglycerolaemia and hyperinsulinaemia in several rodent models of diabetes. In the present study, we have found that this compound increased tyrosine phosphorylation of the IR beta-subunit and IR substrate 1 in primary rat adipocytes as well as induced phosphorylation of Akt, the 70 kDa ribosomal protein S6 kinase and glycogen synthase-3 (deactivation) in Chinese-hamster ovary cells expressing human IR. Similar to insulin, compound 2 stimulated glucose uptake, glycogen synthesis and inhibited isoprenaline-stimulated lipolysis in adipocytes. A structurally related analogue (compound 3) was devoid of the above activities suggesting that the activity of compound 2 is specifically mediated by targeted IR activation. The effects of compound 2 on stimulation of glucose uptake, glycogen synthesis and inhibition of lipolysis were blocked by wortmannin, consistent with the involvement of a phosphoinositide 3-kinase-dependent pathway. In addition, compound 2, but not compound 3, exhibited additive or synergistic effects with sub-maximal concentrations of insulin in rat adipocytes. Thus the IR activator was capable of activating insulin-mediated signalling and metabolic pathways in primary adipocytes. These results demonstrate that IR activators have implications for the future development of new therapeutic approaches to Type I and Type II diabetes.

Full Text

The Full Text of this article is available as a PDF (181.7 KB).

Selected References

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

  1. Agius L., Peak M., Alberti K. G. Regulation of glycogen synthesis from glucose and gluconeogenic precursors by insulin in periportal and perivenous rat hepatocytes. Biochem J. 1990 Feb 15;266(1):91–102. doi: 10.1042/bj2660091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brillon D. J., Freidenberg G. R., Henry R. R., Olefsky J. M. Mechanism of defective insulin-receptor kinase activity in NIDDM. Evidence for two receptor populations. Diabetes. 1989 Mar;38(3):397–403. doi: 10.2337/diab.38.3.397. [DOI] [PubMed] [Google Scholar]
  3. Carlsen J., Christiansen K., Vinten J. Insulin stimulated glycogen synthesis in isolated rat hepatocytes: effect of protein kinase inhibitors. Cell Signal. 1997 Sep;9(6):447–450. doi: 10.1016/s0898-6568(97)00035-1. [DOI] [PubMed] [Google Scholar]
  4. Caro J. F., Sinha M. K., Raju S. M., Ittoop O., Pories W. J., Flickinger E. G., Meelheim D., Dohm G. L. Insulin receptor kinase in human skeletal muscle from obese subjects with and without noninsulin dependent diabetes. J Clin Invest. 1987 May;79(5):1330–1337. doi: 10.1172/JCI112958. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Castan I., Wijkander J., Manganiello V., Degerman E. Mechanisms of inhibition of lipolysis by insulin, vanadate and peroxovanadate in rat adipocytes. Biochem J. 1999 Apr 15;339(Pt 2):281–289. [PMC free article] [PubMed] [Google Scholar]
  6. Cheatham B., Kahn C. R. Insulin action and the insulin signaling network. Endocr Rev. 1995 Apr;16(2):117–142. doi: 10.1210/edrv-16-2-117. [DOI] [PubMed] [Google Scholar]
  7. Cheatham B., Vlahos C. J., Cheatham L., Wang L., Blenis J., Kahn C. R. Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation. Mol Cell Biol. 1994 Jul;14(7):4902–4911. doi: 10.1128/mcb.14.7.4902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cong L. N., Chen H., Li Y., Zhou L., McGibbon M. A., Taylor S. I., Quon M. J. Physiological role of Akt in insulin-stimulated translocation of GLUT4 in transfected rat adipose cells. Mol Endocrinol. 1997 Dec;11(13):1881–1890. doi: 10.1210/mend.11.13.0027. [DOI] [PubMed] [Google Scholar]
  9. Cross D. A., Alessi D. R., Cohen P., Andjelkovich M., Hemmings B. A. Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature. 1995 Dec 21;378(6559):785–789. doi: 10.1038/378785a0. [DOI] [PubMed] [Google Scholar]
  10. Dozio N., Micossi P., Galimberti G., Sartori S., Pozza G., Dosio F., Savi A., Gerundini P. G., Fazio F., Chiumello G. In vivo demonstration of insulin-receptor defect with 123I-labeled insulin and scintigraphic scanning in severe insulin resistance. Diabetes Care. 1992 May;15(5):651–656. doi: 10.2337/diacare.15.5.651. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Goodyear L. J., Giorgino F., Sherman L. A., Carey J., Smith R. J., Dohm G. L. Insulin receptor phosphorylation, insulin receptor substrate-1 phosphorylation, and phosphatidylinositol 3-kinase activity are decreased in intact skeletal muscle strips from obese subjects. J Clin Invest. 1995 May;95(5):2195–2204. doi: 10.1172/JCI117909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodyear L. J., Giorgino F., Sherman L. A., Carey J., Smith R. J., Dohm G. L. Insulin receptor phosphorylation, insulin receptor substrate-1 phosphorylation, and phosphatidylinositol 3-kinase activity are decreased in intact skeletal muscle strips from obese subjects. J Clin Invest. 1995 May;95(5):2195–2204. doi: 10.1172/JCI117909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hajduch E., Alessi D. R., Hemmings B. A., Hundal H. S. Constitutive activation of protein kinase B alpha by membrane targeting promotes glucose and system A amino acid transport, protein synthesis, and inactivation of glycogen synthase kinase 3 in L6 muscle cells. Diabetes. 1998 Jul;47(7):1006–1013. doi: 10.2337/diabetes.47.7.1006. [DOI] [PubMed] [Google Scholar]
  15. Hara K., Yonezawa K., Sakaue H., Ando A., Kotani K., Kitamura T., Kitamura Y., Ueda H., Stephens L., Jackson T. R. 1-Phosphatidylinositol 3-kinase activity is required for insulin-stimulated glucose transport but not for RAS activation in CHO cells. Proc Natl Acad Sci U S A. 1994 Aug 2;91(16):7415–7419. doi: 10.1073/pnas.91.16.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kahn B. B., Simpson I. A., Cushman S. W. Divergent mechanisms for the insulin resistant and hyperresponsive glucose transport in adipose cells from fasted and refed rats. Alterations in both glucose transporter number and intrinsic activity. J Clin Invest. 1988 Aug;82(2):691–699. doi: 10.1172/JCI113649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kahn C. R. Banting Lecture. Insulin action, diabetogenes, and the cause of type II diabetes. Diabetes. 1994 Aug;43(8):1066–1084. doi: 10.2337/diab.43.8.1066. [DOI] [PubMed] [Google Scholar]
  18. Kasuga M., Fujita-Yamaguchi Y., Blithe D. L., Kahn C. R. Tyrosine-specific protein kinase activity is associated with the purified insulin receptor. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2137–2141. doi: 10.1073/pnas.80.8.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kerouz N. J., Hörsch D., Pons S., Kahn C. R. Differential regulation of insulin receptor substrates-1 and -2 (IRS-1 and IRS-2) and phosphatidylinositol 3-kinase isoforms in liver and muscle of the obese diabetic (ob/ob) mouse. J Clin Invest. 1997 Dec 15;100(12):3164–3172. doi: 10.1172/JCI119872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Le Marchand-Brustel Y., Gautier N., Cormont M., Van Obberghen E. Wortmannin inhibits the action of insulin but not that of okadaic acid in skeletal muscle: comparison with fat cells. Endocrinology. 1995 Aug;136(8):3564–3570. doi: 10.1210/endo.136.8.7628394. [DOI] [PubMed] [Google Scholar]
  21. Li J., Elberg G., Sekar N., bin He Z., Shechter Y. Antilipolytic actions of vanadate and insulin in rat adipocytes mediated by distinctly different mechanisms. Endocrinology. 1997 Jun;138(6):2274–2279. doi: 10.1210/endo.138.6.5158. [DOI] [PubMed] [Google Scholar]
  22. Liu K., Xu L., Szalkowski D., Li Z., Ding V., Kwei G., Huskey S., Moller D. E., Heck J. V., Zhang B. B. Discovery of a potent, highly selective, and orally efficacious small-molecule activator of the insulin receptor. J Med Chem. 2000 Sep 21;43(19):3487–3494. doi: 10.1021/jm000285q. [DOI] [PubMed] [Google Scholar]
  23. Michael M. D., Kulkarni R. N., Postic C., Previs S. F., Shulman G. I., Magnuson M. A., Kahn C. R. Loss of insulin signaling in hepatocytes leads to severe insulin resistance and progressive hepatic dysfunction. Mol Cell. 2000 Jul;6(1):87–97. [PubMed] [Google Scholar]
  24. Mooney R. A., Ebersohl R. D., McDonald J. M. Insulin-mediated antilipolysis in permeabilized rat adipocytes. J Biol Chem. 1984 Jun 25;259(12):7701–7704. [PubMed] [Google Scholar]
  25. Morimoto C., Tsujita T., Okuda H. Antilipolytic actions of insulin on basal and hormone-induced lipolysis in rat adipocytes. J Lipid Res. 1998 May;39(5):957–962. [PubMed] [Google Scholar]
  26. Mèndez R., Myers M. G., Jr, White M. F., Rhoads R. E. Stimulation of protein synthesis, eukaryotic translation initiation factor 4E phosphorylation, and PHAS-I phosphorylation by insulin requires insulin receptor substrate 1 and phosphatidylinositol 3-kinase. Mol Cell Biol. 1996 Jun;16(6):2857–2864. doi: 10.1128/mcb.16.6.2857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nikoulina S. E., Ciaraldi T. P., Mudaliar S., Mohideen P., Carter L., Henry R. R. Potential role of glycogen synthase kinase-3 in skeletal muscle insulin resistance of type 2 diabetes. Diabetes. 2000 Feb;49(2):263–271. doi: 10.2337/diabetes.49.2.263. [DOI] [PubMed] [Google Scholar]
  28. Okada T., Kawano Y., Sakakibara T., Hazeki O., Ui M. Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. J Biol Chem. 1994 Feb 4;269(5):3568–3573. [PubMed] [Google Scholar]
  29. Pessin J. E., Saltiel A. R. Signaling pathways in insulin action: molecular targets of insulin resistance. J Clin Invest. 2000 Jul;106(2):165–169. doi: 10.1172/JCI10582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Qureshi S. A., Ding V., Li Z., Szalkowski D., Biazzo-Ashnault D. E., Xie D., Saperstein R., Brady E., Huskey S., Shen X. Activation of insulin signal transduction pathway and anti-diabetic activity of small molecule insulin receptor activators. J Biol Chem. 2000 Nov 24;275(47):36590–36595. doi: 10.1074/jbc.M006287200. [DOI] [PubMed] [Google Scholar]
  31. Shepherd P. R., Navé B. T., Siddle K. Insulin stimulation of glycogen synthesis and glycogen synthase activity is blocked by wortmannin and rapamycin in 3T3-L1 adipocytes: evidence for the involvement of phosphoinositide 3-kinase and p70 ribosomal protein-S6 kinase. Biochem J. 1995 Jan 1;305(Pt 1):25–28. doi: 10.1042/bj3050025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shepherd P. R., Withers D. J., Siddle K. Phosphoinositide 3-kinase: the key switch mechanism in insulin signalling. Biochem J. 1998 Aug 1;333(Pt 3):471–490. doi: 10.1042/bj3330471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith U., Carvalho E., Mosialou E., Beguinot F., Formisano P., Rondinone C. PKB inhibition prevents the stimulatory effect of insulin on glucose transport and protein translocation but not the antilipolytic effect in rat adipocytes. Biochem Biophys Res Commun. 2000 Feb 16;268(2):315–320. doi: 10.1006/bbrc.2000.2145. [DOI] [PubMed] [Google Scholar]
  34. Sutherland C., Waltner-Law M., Gnudi L., Kahn B. B., Granner D. K. Activation of the ras mitogen-activated protein kinase-ribosomal protein kinase pathway is not required for the repression of phosphoenolpyruvate carboxykinase gene transcription by insulin. J Biol Chem. 1998 Feb 6;273(6):3198–3204. doi: 10.1074/jbc.273.6.3198. [DOI] [PubMed] [Google Scholar]
  35. Taha C., Klip A. The insulin signaling pathway. J Membr Biol. 1999 May 1;169(1):1–12. doi: 10.1007/pl00005896. [DOI] [PubMed] [Google Scholar]
  36. Thomas J. A., Schlender K. K., Larner J. A rapid filter paper assay for UDPglucose-glycogen glucosyltransferase, including an improved biosynthesis of UDP-14C-glucose. Anal Biochem. 1968 Oct 24;25(1):486–499. doi: 10.1016/0003-2697(68)90127-9. [DOI] [PubMed] [Google Scholar]
  37. Tsakiridis T., McDowell H. E., Walker T., Downes C. P., Hundal H. S., Vranic M., Klip A. Multiple roles of phosphatidylinositol 3-kinase in regulation of glucose transport, amino acid transport, and glucose transporters in L6 skeletal muscle cells. Endocrinology. 1995 Oct;136(10):4315–4322. doi: 10.1210/endo.136.10.7664650. [DOI] [PubMed] [Google Scholar]
  38. Valverde A. M., Navarro P., Teruel T., Conejo R., Benito M., Lorenzo M. Insulin and insulin-like growth factor I up-regulate GLUT4 gene expression in fetal brown adipocytes, in a phosphoinositide 3-kinase-dependent manner. Biochem J. 1999 Feb 1;337(Pt 3):397–405. [PMC free article] [PubMed] [Google Scholar]
  39. Van Obberghen E., Ksauga M., Le Cam A., Hedo J. A., Itin A., Harrison L. C. Biosynthetic labeling of insulin receptor: studies of subunits in cultured human IM-9 lymphocytes. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1052–1056. doi: 10.1073/pnas.78.2.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang Q., Somwar R., Bilan P. J., Liu Z., Jin J., Woodgett J. R., Klip A. Protein kinase B/Akt participates in GLUT4 translocation by insulin in L6 myoblasts. Mol Cell Biol. 1999 Jun;19(6):4008–4018. doi: 10.1128/mcb.19.6.4008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. White M. F., Maron R., Kahn C. R. Insulin rapidly stimulates tyrosine phosphorylation of a Mr-185,000 protein in intact cells. Nature. 1985 Nov 14;318(6042):183–186. doi: 10.1038/318183a0. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Zhang B., Salituro G., Szalkowski D., Li Z., Zhang Y., Royo I., Vilella D., Díez M. T., Pelaez F., Ruby C. Discovery of a small molecule insulin mimetic with antidiabetic activity in mice. Science. 1999 May 7;284(5416):974–977. doi: 10.1126/science.284.5416.974. [DOI] [PubMed] [Google Scholar]

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

RESOURCES