Insulin-receptor Phosphotyrosyl-protein Phosphatases

Overview

Journal Biochem J

Specialty Biochemistry

Date 1988 Dec 15

PMID 2852008

Citations 13

Authors

M J King

G J Sale

Affiliations

Soon will be listed here.

Abstract

Calmodulin-dependent protein phosphatase has been proposed to be an important phosphotyrosyl-protein phosphatase. The ability of the enzyme to attack autophosphorylated insulin receptor was examined and compared with the known ability of the enzyme to act on autophosphorylated epidermal-growth-factor (EGF) receptor. Purified calmodulin-dependent protein phosphatase was shown to catalyse the complete dephosphorylation of phosphotyrosyl-(insulin receptor). When compared at similar concentrations, 32P-labelled EGF receptor was dephosphorylated at greater than 3 times the rate of 32P-labelled insulin receptor; both dephosphorylations exhibited similar dependence on metal ions and calmodulin. Native phosphotyrosyl-protein phosphatases in cell extracts were also characterized. With rat liver, heart or brain, most (75%) of the native phosphatase activity against both 32P-labelled insulin and EGF receptors was recovered in the particulate fraction of the cell, with only 25% in the soluble fraction. This subcellular distribution contrasts with results of previous studies using artificial substrates, which found most of the phosphotyrosyl-protein phosphatase activity in the soluble fraction of the cell. Properties of particulate and soluble phosphatase activity against 32P-labelled insulin and EGF receptors are reported. The contribution of calmodulin-dependent protein phosphatase activity to phosphotyrosyl-protein phosphatase activity in cell fractions was determined by utilizing the unique metal-ion dependence of calmodulin-dependent protein phosphatase. Whereas Ni2+ (1 mM) markedly activated the calmodulin-dependent protein phosphatase, it was found to inhibit potently both particulate and soluble phosphotyrosyl-protein phosphatase activity. In fractions from rat liver, brain and heart, total phosphotyrosyl-protein phosphatase activity against both 32P-labelled receptors was inhibited by 99.5 +/- 6% (mean +/- S.E.M., 30 observations) by Ni2+. Results of Ni2+ inhibition studies were confirmed by other methods. It is concluded that in cell extracts phosphotyrosyl-protein phosphatases other than calmodulin-dependent protein phosphatase are the major phosphotyrosyl-(insulin receptor) and -(EGF receptor) phosphatases.

Citing Articles

Insulin receptor-associated protein tyrosine phosphatase(s): role in insulin action.

Drake P, Posner B Mol Cell Biochem. 1998; 182(1-2):79-89.

PMID: 9609117

Insulin receptor internalization and signalling.

Di Guglielmo G, Drake P, Baass P, Authier F, Posner B, Bergeron J Mol Cell Biochem. 1998; 182(1-2):59-63.

PMID: 9609114

Nitric oxide reversibly inhibits the epidermal growth factor receptor tyrosine kinase.

Estrada C, Gomez C, Martin-Nieto J, de Frutos T, Jimenez A, Villalobo A Biochem J. 1997; 326 ( Pt 2):369-76.

PMID: 9291107 PMC: 1218680. DOI: 10.1042/bj3260369.

Studies on an insulin-stimulated insulin receptor serine kinase activity: separation of the kinase activity from the insulin receptor and its reconstitution back to the insulin receptor.

Asamoah K, Atkinson P, Carter W, Sale G Biochem J. 1995; 308 ( Pt 3):915-22.

PMID: 8948451 PMC: 1136811. DOI: 10.1042/bj3080915.

In vivo modulation of N-myristoyltransferase activity by orthovanadate.

King M, Pugazhenthi S, Khandelwal R, Sharma R Mol Cell Biochem. 1995; 153(1-2):151-5.

PMID: 8927031 DOI: 10.1007/BF01075931.

References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Sale G . Recent progress in our understanding of the mechanism of action of insulin. Int J Biochem. 1988; 20(9):897-908. DOI: 10.1016/0020-711x(88)90173-5. View

Brautigan D, Bornstein P, Gallis B . Phosphotyrosyl-protein phosphatase. Specific inhibition by Zn. J Biol Chem. 1981; 256(13):6519-22. View

Swarup G, Cohen S, Garbers D . Selective dephosphorylation of proteins containing phosphotyrosine by alkaline phosphatases. J Biol Chem. 1981; 256(15):8197-201. View

Foulkes J, Howard R, Ziemiecki A . Detection of a novel mammalian protein phosphatase with activity for phosphotyrosine. FEBS Lett. 1981; 130(2):197-200. DOI: 10.1016/0014-5793(81)81118-0. View

Stewart A, Ingebritsen T, Manalan A, Klee C, Cohen P . Discovery of a Ca2+- and calmodulin-dependent protein phosphatase: probable identity with calcineurin (CaM-BP80). FEBS Lett. 1982; 137(1):80-4. DOI: 10.1016/0014-5793(82)80319-0. View

Kasuga M, Zick Y, Blithe D, Crettaz M, Kahn C . Insulin stimulates tyrosine phosphorylation of the insulin receptor in a cell-free system. Nature. 1982; 298(5875):667-9. DOI: 10.1038/298667a0. View

Swarup G, Cohen S, Garbers D . Inhibition of membrane phosphotyrosyl-protein phosphatase activity by vanadate. Biochem Biophys Res Commun. 1982; 107(3):1104-9. DOI: 10.1016/0006-291x(82)90635-0. View

Leis J, Kaplan N . An acid phosphatase in the plasma membranes of human astrocytoma showing marked specificity toward phosphotyrosine protein. Proc Natl Acad Sci U S A. 1982; 79(21):6507-11. PMC: 347156. DOI: 10.1073/pnas.79.21.6507. View

10.

Foulkes J, Erikson E, Erikson R . Separation of multiple phosphotyrosyl-and phosphoseryl-protein phosphatases from chicken brain. J Biol Chem. 1983; 258(1):431-8. View

11.

Stewart A, Ingebritsen T, Cohen P . The protein phosphatases involved in cellular regulation. 5. Purification and properties of a Ca2+/calmodulin-dependent protein phosphatase (2B) from rabbit skeletal muscle. Eur J Biochem. 1983; 132(2):289-95. DOI: 10.1111/j.1432-1033.1983.tb07361.x. View

12.

Ingebritsen T, Stewart A, Cohen P . The protein phosphatases involved in cellular regulation. 6. Measurement of type-1 and type-2 protein phosphatases in extracts of mammalian tissues; an assessment of their physiological roles. Eur J Biochem. 1983; 132(2):297-307. DOI: 10.1111/j.1432-1033.1983.tb07362.x. View

13.

Rosen O, Herrera R, Olowe Y, Petruzzelli L, Cobb M . Phosphorylation activates the insulin receptor tyrosine protein kinase. Proc Natl Acad Sci U S A. 1983; 80(11):3237-40. PMC: 394015. DOI: 10.1073/pnas.80.11.3237. View

14.

Pallen C, Wang J . Calmodulin-stimulated dephosphorylation of p-nitrophenyl phosphate and free phosphotyrosine by calcineurin. J Biol Chem. 1983; 258(14):8550-3. View

15.

KING M, Huang C . Activation of calcineurin by nickel ions. Biochem Biophys Res Commun. 1983; 114(3):955-61. DOI: 10.1016/0006-291x(83)90653-8. View

16.

Tallant E, Cheung W . Calmodulin-dependent protein phosphatase: a developmental study. Biochemistry. 1983; 22(15):3630-5. DOI: 10.1021/bi00284a014. View

17.

Chernoff J, Li H . Multiple forms of phosphotyrosyl- and phosphoseryl-protein phosphatase from cardiac muscle: partial purification and characterization of an EDTA-stimulated phosphotyrosyl-protein phosphatase. Arch Biochem Biophys. 1983; 226(2):517-30. DOI: 10.1016/0003-9861(83)90321-1. View

18.

Klee C, KRINKS M, Manalan A, Cohen P, Stewart A . Isolation and characterization of bovine brain calcineurin: a calmodulin-stimulated protein phosphatase. Methods Enzymol. 1983; 102:227-44. DOI: 10.1016/s0076-6879(83)02024-8. View

19.

Tallant E, Wallace R, Cheung W . Purification and radioimmunoassay of calmodulin-dependent protein phosphatase from bovine brain. Methods Enzymol. 1983; 102:244-56. DOI: 10.1016/s0076-6879(83)02025-x. View

20.

Li H, Chernoff J, Chen L, Kirschonbaum A . A phosphotyrosyl-protein phosphatase activity associated with acid phosphatase from human prostate gland. Eur J Biochem. 1984; 138(1):45-51. DOI: 10.1111/j.1432-1033.1984.tb07879.x. View