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. 1990 Jun;85(6):1754–1760. doi: 10.1172/JCI114632

Hepatic protein phosphotyrosine phosphatase. Dephosphorylation of insulin and epidermal growth factor receptors in normal and alloxan diabetic rats.

P A Gruppuso 1, J M Boylan 1, B I Posner 1, R Faure 1, D L Brautigan 1
PMCID: PMC296637  PMID: 2161429

Abstract

Polypeptide hormone signal transmission by receptor tyrosine kinases requires the rapid reversal of tyrosine phosphorylation by protein phosphotyrosine phosphatases (PPTPases). We studied hepatic PPTPases in the rat with emphasis on acute and chronic regulation by insulin. PPTPase activity with artificial substrates ([32P]Tyr-reduced, carboxyamidomethylated, and maleylated lysozyme and [32P]Tyr-poly[glutamic acid:tyrosine] 4:1) was present in distinct membrane, cytoskeletal, and cytosolic fractions. These PPTPase activities were unaffected by alloxan diabetes. Acute administration of insulin to normal animals also did not change PPTPase activity in liver plasma membranes or endosomal membranes. Although alloxan diabetes did not affect PPTPase activity measured with artificial substrates or with epidermal growth factor receptors, a decrease in insulin receptor dephosphorylation was noted. Dephosphorylation of hepatic receptors from normal and diabetic rats by membrane PPTPase from control rats was similar. These results indicate that alloxan diabetes does not lead to a generalized effect on hepatic PPTPase activity, although a substrate-specific decrease in activity with the insulin receptor may occur.

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Selected References

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

  1. Alemany S., Pelech S., Brierley C. H., Cohen P. The protein phosphatases involved in cellular regulation. Evidence that dephosphorylation of glycogen phosphorylase and glycogen synthase in the glycogen and microsomal fractions of rat liver are catalysed by the same enzyme: protein phosphatase-1. Eur J Biochem. 1986 Apr 1;156(1):101–110. doi: 10.1111/j.1432-1033.1986.tb09554.x. [DOI] [PubMed] [Google Scholar]
  2. Amatruda J. M., Roncone A. M. Normal hepatic insulin receptor autophosphorylation in nonketotic diabetes mellitus. Biochem Biophys Res Commun. 1985 May 31;129(1):163–170. doi: 10.1016/0006-291x(85)91417-2. [DOI] [PubMed] [Google Scholar]
  3. BERGMEYER H. U., BERNT E. ENZYMATISCHE BESTIMMUNG VON KETON-KOERPERN IM BLUT. Enzymol Biol Clin (Basel) 1965;19:65–76. [PubMed] [Google Scholar]
  4. Bernier M., Laird D. M., Lane M. D. Insulin-activated tyrosine phosphorylation of a 15-kilodalton protein in intact 3T3-L1 adipocytes. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1844–1848. doi: 10.1073/pnas.84.7.1844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Blackshear P. J., Nemenoff R. A., Avruch J. Characteristics of insulin and epidermal growth factor stimulation of receptor autophosphorylation in detergent extracts of rat liver and transplantable rat hepatomas. Endocrinology. 1984 Jan;114(1):141–152. doi: 10.1210/endo-114-1-141. [DOI] [PubMed] [Google Scholar]
  6. Bollen M., Stalmans W. The hepatic defect in glycogen synthesis in chronic diabetes involves the G-component of synthase phosphatase. Biochem J. 1984 Jan 15;217(2):427–434. doi: 10.1042/bj2170427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burant C. F., Treutelaar M. K., Buse M. G. Diabetes-induced functional and structural changes in insulin receptors from rat skeletal muscle. J Clin Invest. 1986 Jan;77(1):260–270. doi: 10.1172/JCI112285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Caro J. F., Ittoop O., Pories W. J., Meelheim D., Flickinger E. G., Thomas F., Jenquin M., Silverman J. F., Khazanie P. G., Sinha M. K. Studies on the mechanism of insulin resistance in the liver from humans with noninsulin-dependent diabetes. Insulin action and binding in isolated hepatocytes, insulin receptor structure, and kinase activity. J Clin Invest. 1986 Jul;78(1):249–258. doi: 10.1172/JCI112558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Carpenter G. Receptors for epidermal growth factor and other polypeptide mitogens. Annu Rev Biochem. 1987;56:881–914. doi: 10.1146/annurev.bi.56.070187.004313. [DOI] [PubMed] [Google Scholar]
  11. Comi R. J., Grunberger G., Gorden P. Relationship of insulin binding and insulin-stimulated tyrosine kinase activity is altered in type II diabetes. J Clin Invest. 1987 Feb;79(2):453–462. doi: 10.1172/JCI112833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. DeFronzo R. A., Hendler R., Simonson D. Insulin resistance is a prominent feature of insulin-dependent diabetes. Diabetes. 1982 Sep;31(9):795–801. doi: 10.2337/diab.31.9.795. [DOI] [PubMed] [Google Scholar]
  13. Desbuquois B., Aurbach G. D. Use of polyethylene glycol to separate free and antibody-bound peptide hormones in radioimmunoassays. J Clin Endocrinol Metab. 1971 Nov;33(5):732–738. doi: 10.1210/jcem-33-5-732. [DOI] [PubMed] [Google Scholar]
  14. Fava R. A., Cohen S. Isolation of a calcium-dependent 35-kilodalton substrate for the epidermal growth factor receptor/kinase from A-431 cells. J Biol Chem. 1984 Feb 25;259(4):2636–2645. [PubMed] [Google Scholar]
  15. Freidenberg G. R., Henry R. R., Klein H. H., Reichart D. R., Olefsky J. M. Decreased kinase activity of insulin receptors from adipocytes of non-insulin-dependent diabetic subjects. J Clin Invest. 1987 Jan;79(1):240–250. doi: 10.1172/JCI112789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ginsberg H., Kimmerling G., Olefsky J. M., Reaven G. M. Demonstration of insulin resistance in untreated adult onset diabetic subjects with fasting hyperglycemia. J Clin Invest. 1975 Mar;55(3):454–461. doi: 10.1172/JCI107951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jones S. W., Erikson R. L., Ingebritsen V. M., Ingebritsen T. S. Phosphotyrosyl-protein phosphatases. I. Separation of multiple forms from bovine brain and purification of the major form to near homogeneity. J Biol Chem. 1989 May 5;264(13):7747–7753. [PubMed] [Google Scholar]
  18. Kadowaki T., Kasuga M., Akanuma Y., Ezaki O., Takaku F. Decreased autophosphorylation of the insulin receptor-kinase in streptozotocin-diabetic rats. J Biol Chem. 1984 Nov 25;259(22):14208–14216. [PubMed] [Google Scholar]
  19. Kasuga M., Karlsson F. A., Kahn C. R. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor. Science. 1982 Jan 8;215(4529):185–187. doi: 10.1126/science.7031900. [DOI] [PubMed] [Google Scholar]
  20. Khan M. N., Savoie S., Bergeron J. J., Posner B. I. Characterization of rat liver endosomal fractions. In vivo activation of insulin-stimulable receptor kinase in these structures. J Biol Chem. 1986 Jun 25;261(18):8462–8472. [PubMed] [Google Scholar]
  21. King M. J., Sale G. J. Assay of phosphotyrosyl protein phosphatase using synthetic peptide 1142-1153 of the insulin receptor. FEBS Lett. 1988 Sep 12;237(1-2):137–140. doi: 10.1016/0014-5793(88)80187-x. [DOI] [PubMed] [Google Scholar]
  22. Kolterman O. G., Insel J., Saekow M., Olefsky J. M. Mechanisms of insulin resistance in human obesity: evidence for receptor and postreceptor defects. J Clin Invest. 1980 Jun;65(6):1272–1284. doi: 10.1172/JCI109790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Margolis R. N., Taylor S. I., Seminara D., Hubbard A. L. Identification of pp120, an endogenous substrate for the hepatocyte insulin receptor tyrosine kinase, as an integral membrane glycoprotein of the bile canalicular domain. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7256–7259. doi: 10.1073/pnas.85.19.7256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Meyerovitch J., Backer J. M., Kahn C. R. Hepatic phosphotyrosine phosphatase activity and its alterations in diabetic rats. J Clin Invest. 1989 Sep;84(3):976–983. doi: 10.1172/JCI114261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Morrison D. K., Kaplan D. R., Escobedo J. A., Rapp U. R., Roberts T. M., Williams L. T. Direct activation of the serine/threonine kinase activity of Raf-1 through tyrosine phosphorylation by the PDGF beta-receptor. Cell. 1989 Aug 25;58(4):649–657. doi: 10.1016/0092-8674(89)90100-1. [DOI] [PubMed] [Google Scholar]
  26. O'Brien R., Houslay M. D., Brindle N. P., Milligan G., Whittaker J., Siddle K. Binding to GDP-agarose identifies a novel 60kDa substrate for the insulin receptor tyrosyl kinase in mouse NIH-3T3 cells expressing high concentrations of the human insulin receptor. Biochem Biophys Res Commun. 1989 Feb 15;158(3):743–748. doi: 10.1016/0006-291x(89)92784-8. [DOI] [PubMed] [Google Scholar]
  27. Okada M., Owada K., Nakagawa H. [Phosphotyrosine]protein phosphatase in rat brain. A major [phosphotyrosine]protein phosphatase is a 23 kDa protein distinct from acid phosphatase. Biochem J. 1986 Oct 1;239(1):155–162. doi: 10.1042/bj2390155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pallen C. J., Panayotou G. N., Sahlin L., Waterfield M. D. Purification of a phosphotyrosine phosphatase that dephosphorylates the epidermal growth factor receptor autophosphorylation sites. Ann N Y Acad Sci. 1988;551:299–308. doi: 10.1111/j.1749-6632.1988.tb22356.x. [DOI] [PubMed] [Google Scholar]
  29. Rees-Jones R. W., Taylor S. I. An endogenous substrate for the insulin receptor-associated tyrosine kinase. J Biol Chem. 1985 Apr 10;260(7):4461–4467. [PubMed] [Google Scholar]
  30. Roome J., O'Hare T., Pilch P. F., Brautigan D. L. Protein phosphotyrosine phosphatase purified from the particulate fraction of human placenta dephosphorylates insulin and growth-factor receptors. Biochem J. 1988 Dec 1;256(2):493–500. doi: 10.1042/bj2560493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rotenberg S. A., Brautigan D. L. Membrane protein phosphotyrosine phosphatase in rabbit kidney. Proteolysis activates the enzyme and generates soluble catalytic fragments. Biochem J. 1987 May 1;243(3):747–754. doi: 10.1042/bj2430747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Shriner C. L., Brautigan D. L. Cytosolic protein phosphotyrosine phosphatases from rabbit kidney. Purification of two distinct enzymes that bind to Zn2+-iminodiacetate agarose. J Biol Chem. 1984 Sep 25;259(18):11383–11390. [PubMed] [Google Scholar]
  33. Sibley D. R., Benovic J. L., Caron M. G., Lefkowitz R. J. Regulation of transmembrane signaling by receptor phosphorylation. Cell. 1987 Mar 27;48(6):913–922. doi: 10.1016/0092-8674(87)90700-8. [DOI] [PubMed] [Google Scholar]
  34. Sinha M. K., Pories W. J., Flickinger E. G., Meelheim D., Caro J. F. Insulin-receptor kinase activity of adipose tissue from morbidly obese humans with and without NIDDM. Diabetes. 1987 May;36(5):620–625. doi: 10.2337/diab.36.5.620. [DOI] [PubMed] [Google Scholar]
  35. Sparks J. W., Brautigan D. L. Specificity of protein phosphotyrosine phosphatases. Comparison with mammalian alkaline phosphatase using polypeptide substrates. J Biol Chem. 1985 Feb 25;260(4):2042–2045. [PubMed] [Google Scholar]
  36. Strout H. V., Vicario P. P., Saperstein R., Slater E. E. A protein phosphotyrosine phosphatase distinct from alkaline phosphatase with activity against the insulin receptor. Biochem Biophys Res Commun. 1988 Mar 15;151(2):633–640. doi: 10.1016/s0006-291x(88)80328-0. [DOI] [PubMed] [Google Scholar]
  37. Tan A. W., Nuttall F. Q. Regulation of synthase phosphatase and phosphorylase phosphatase in rat liver. Biochim Biophys Acta. 1976 Aug 12;445(1):118–130. doi: 10.1016/0005-2744(76)90165-0. [DOI] [PubMed] [Google Scholar]
  38. Tonks N. K., Diltz C. D., Fischer E. H. Characterization of the major protein-tyrosine-phosphatases of human placenta. J Biol Chem. 1988 May 15;263(14):6731–6737. [PubMed] [Google Scholar]
  39. Tonks N. K., Diltz C. D., Fischer E. H. Purification of the major protein-tyrosine-phosphatases of human placenta. J Biol Chem. 1988 May 15;263(14):6722–6730. [PubMed] [Google Scholar]
  40. Wahl M. I., Nishibe S., Suh P. G., Rhee S. G., Carpenter G. Epidermal growth factor stimulates tyrosine phosphorylation of phospholipase C-II independently of receptor internalization and extracellular calcium. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1568–1572. doi: 10.1073/pnas.86.5.1568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Watarai T., Kobayashi M., Takata Y., Sasaoka T., Iwasaki M., Shigeta Y. Alteration of insulin-receptor kinase activity by high-fat feeding. Diabetes. 1988 Oct;37(10):1397–1404. doi: 10.2337/diab.37.10.1397. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Williams L. T., Tremble P. M., Lavin M. F., Sunday M. E. Platelet-derived growth factor receptors form a high affinity state in membrane preparations. Kinetics and affinity cross-linking studies. J Biol Chem. 1984 Apr 25;259(8):5287–5294. [PubMed] [Google Scholar]
  44. Yarden Y., Ullrich A. Growth factor receptor tyrosine kinases. Annu Rev Biochem. 1988;57:443–478. doi: 10.1146/annurev.bi.57.070188.002303. [DOI] [PubMed] [Google Scholar]

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