Advertisement
Journal of Biological Chemistry
Open access logo
Advanced SearchSave search
Full length article| Volume 264, ISSUE 18, P10828-10835, June 25, 1989

Mechanism of Phosphorylation of the Epidermal Growth Factor Receptor at Threonine 669

Open AccessPublished:June 25, 1989DOI:https://doi.org/10.1016/S0021-9258(18)81695-7
Mechanism of Phosphorylation of the Epidermal Growth Factor Receptor at Threonine 669
Previous ArticleEvidence for Multiple, ras-like, Guanine Nucleotide-binding Proteins in Swiss 3T3 Plasma Membranes
Next ArticleT Cell Activation Induces Rapid Tyrosine Phosphorylation of a Limited Number of Cellular Substrates
      This paper is only available as a PDF. To read, Please Download here.
      The major site of phosphorylation of the epidermal growth factor (EGF) receptor after treatment of cells with EGF is threonine 669. Phosphorylation of this site is also associated with the transmodulation of the EGF receptor caused by platelet-derived growth factor and phorbol ester. A distinctive feature of the primary sequence surrounding threonine 669 is the proximity of 2 proline residues (-Pro-Leu-Thr689-Pro-). This site is not a substrate for phosphorylation by protein kinase C. To investigate the mechanism of the increased phosphorylation of the EGF receptor at threonine 669, in vitro assays were used to measure protein kinase and protein phosphatase activities present in homogenates prepared from cells treated with and without EGF. No evidence for the regulation of protein phosphatase activity was obtained in experiments using the [32P]phosphate-labeled EGF receptor as a substrate. A synthetic peptide corresponding to residues 663–681 of the EGF receptor was used as a substrate for protein kinase assays. Incubation of murine 3T3 L1 pre-adipocytes and human WI-38 fibroblasts with EGF caused a rapid increase (3–10-fold) in the level of threonine protein kinase activity detected in cell homogenates. Similar results were obtained after EGF treatment of Chinese hamster ovary cells expressing wild-type (Thr669) and mutated (Ala669) human EGF receptors. Activation of the threonine protein kinase activity was also observed in cells treated with platelet-derived growth factor, serum, and phorbol ester. Insulin-like growth factor-1 caused no significant change in protein kinase activity. Together these data indicate a role for the regulation of the activity of a threonine protein kinase in the control of the phosphorylation state of the EGF receptor at threonine 669. The significance of the identification of a growth factor-stimulated threonine protein kinase to the mechanism of signal transduction is discussed.

      REFERENCES

        • Zoller M.J.
        • Smith M.
        DNA (NY). 1984; 3: 479-488
        • Davis R.J.
        • Meisner H.
        J. Biol. Chem. 1987; 262: 16041-16047
        • Cepko C.J.
        • Roberts B.E.
        • Mulligan R.C.
        Cell. 1984; 37: 1053-1062
        • Beemon K.
        • Hunter T.
        J. Virol. 1978; 28: 551-556
        • Hunter T.
        • Sefton B.M.
        Proc. Natl. Acad. Sci. U. S. A. 1980; 77: 1311-1315
        • Zimmerman C.L.
        • Appella E.
        • Pisano J.J.
        Anal. Biochem. 1976; 75: 75-85
        • Ullrich A.
        • Coussens L.
        • Mayflick J.S.
        • Dull T.J.
        • Gray A.
        • Tam A.W.
        • Lee J.
        • Yarden Y.
        • Libermann T.A.
        • Schlessinger J.
        • Downward J.
        • Mayes E.L.V.
        • Whittle N.
        • Waterfield M.D.
        • Seeburg P.H.
        Nature. 1984; 309: 418-425
        • Scatchard G.
        Ann. N. Y. Acad. Sci. 1949; 51: 660-672
        • Munson P.J.
        • Rodbard D.
        Anal. Biochem. 1980; 107: 220-239
        • Hunter T.
        • Cooper J.A.
        Cell. 1981; 24: 741-752
        • Chan C.P.
        • McNall S.J.
        • Krebs E.G.
        • Fischer E.H.
        Proc. Natl. Acad. Sci. U. S. A. 1988; 85: 6257-6261
        • Edelman A.M.
        • Blumenthal D.K.
        • Krebs E.G.
        Annu. Rev. Biochem. 1987; 56: 567-613
        • Woodgett J.R.
        • Cohen P.
        Biochim. Biophys. Acta. 1984; 788: 339-347
        • Ramakrishna S.
        • Benjamin W.B.
        J. Biol. Chem. 1985; 260: 12280-12286
        • Sheorain V.S.
        • Ramakrishna S.
        • Benjamin W.B.
        • Soderling T.R.
        J. Biol. Chem. 1985; 260: 12287-12292
        • Taniuchi M.
        • Johnson Jr., E.M.
        • Roach P.J.
        • Lawrence Jr., J.C.
        J. Biol. Chem. 1986; 261: 13342-13349
        • Fiol C.J.
        • Mahrenholz A.M.
        • Wang Y.
        • Roeske R.W.
        • Roach P.J.
        J. Biol. Chem. 1987; 262: 14042-14048
        • Ramakrishna S.
        • Benjamin W.B.
        J. Biol. Chem. 1988; 263: 12677-12681
        • Giugni T.D.
        • Chen K.
        • Cohen S.
        J. Biol. Chem. 1988; 263: 18988-18995
        • Schlessinger J.
        Biochemistry. 1988; 27: 3119-3123
        • Davis R.J.
        • Czech M.P.
        Cancer Cells. 1985; 3: 101-108
        • Hunter T.
        • Ling N.
        • Cooper J.A.
        Nature. 1984; 311: 480-483
        • Davis R.J.
        • Czech M.P.
        Proc. Natl. Acad. Sci. U. S. A. 1985; 82: 1974-1978
        • Davis R.J.
        J. Biol. Chem. 1988; 263: 9462-9469
        • Livneh E.
        • Dull T.J.
        • Berent E.
        • Prywes R.
        • Ullrich A.
        • Schlessinger J.
        Mol. Cell. Biol. 1988; 8: 2302-2308
        • Lin C.R.
        • Chen S.W.
        • Lazar C.S.
        • Carpenter C.D.
        • Gill G.N.
        • Evans R.N.
        • Rosenfeld M.G.
        Cell. 1986; 44: 839-846
        • Cochet C.
        • Gill G.N.
        • Meisenhelder J.
        • Cooper J.A.
        • Hunter T.
        J. Biol. Chem. 1984; 259: 2553-2558
        • Downward J.
        • Waterfield M.D.
        • Parker P.J.
        J. Biol. Chem. 1985; 260: 14538-14546
        • Iwashita S.
        • Fox C.F.
        J. Biol. Chem. 1984; 259: 2559-2567
        • Decker S.
        Mol. Cell. Biol. 1984; 4: 1718-1723
        • Davis R.J.
        • Ganong B.R.
        • Bell R.M.
        • Czech M.P.
        J. Biol. Chem. 1985; 260: 1562-1566
        • Davis R.J.
        • Ganong B.R.
        • Bell R.M.
        • Czech M.P.
        J. Biol. Chem. 1985; 260: 5315-5322
        • Davis R.J.
        • Czech M.P.
        Proc. Natl. Acad. Sci. U. S. A. 1985; 82: 4080-4084
        • Davis R.J.
        • Czech M.P.
        J. Biol. Chem. 1987; 262: 6832-6841
        • Heisermann G.J.
        • Gill G.N.
        J. Biol. Chem. 1988; 263: 13152-13158
        • Savage Jr., C.R.
        • Cohen S.
        J. Biol. Chem. 1972; 247: 7609-7611
        • Matrisian L.M.
        • Larsen B.R.
        • Finch J.S.
        • Magun B.E.
        Anal. Biochem. 1982; 125: 339-351
        • Pessin J.E.
        • Gitomer W.
        • Oka Y.
        • Oppenheimer C.L.
        • Czech M.P.
        J. Biol. Chem. 1983; 258: 7386-7394

      Article Info

      Publication History

      Published online: June 25, 1989
      Received: February 2, 1989

      Identification

      DOI: https://doi.org/10.1016/S0021-9258(18)81695-7

      Copyright

      © 1989 ASBMB. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology.

      User License

      Creative Commons Attribution (CC BY 4.0) |
      How you can reuse Information Icon
      Creative Commons Attribution (CC BY 4.0)

      Permitted

      • Read, print & download
      • Redistribute or republish the final article
      • Text & data mine
      • Translate the article
      • Reuse portions or extracts from the article in other works
      • Sell or re-use for commercial purposes

      Elsevier's open access license policy

      ScienceDirect

      Access this article on ScienceDirect

      ASBMB  ASBMB  ASBMB  ASBMB

      ISSN 0021-9258
      We use cookies to help provide and enhance our service and tailor content. To update your cookie settings, please visit the for this site.
      Copyright © 2022 Elsevier Inc. except certain content provided by third parties. The content on this site is intended for healthcare professionals.


      RELX