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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1982 Jul;79(14):4303–4307. doi: 10.1073/pnas.79.14.4303

Platelet-derived growth factor stimulates tyrosine-specific protein kinase activity in Swiss mouse 3T3 cell membranes.

J Nishimura, J S Huang, T F Deuel
PMCID: PMC346659  PMID: 6181505

Abstract

Platelet-derived growth factor (PDGF) stimulates the incorporation of 32P from [gamma-32P]ATP into a Mr approximately 170,000 protein by an endogenous tyrosine-specific protein kinase in membrane preparations of Swiss mouse 3T3 cells. Epidermal growth factor (EGF), but not fibroblast growth factor (FGF) or insulin, stimulates limited incorporation of 32P into a protein of similar molecular weight. The ligand concentration required for half-maximal activity (S0.5) for PDGF stimulation of phosphorylation is 50 ng/ml; saturation is achieved at 300 ng/ml. The S0.5 for ATP is 15 microM. Mg2+ or Mn2+ is required for protein kinase activity. Stimulation of PDGF results in the preferential phosphorylation of tyrosine residues in this Mr approximately 170,000 membrane protein. The Mr approximately 170,000 protein can be resolved into Mr approximately 180,000 and 160,000 components in 4% NaDodSO4 gels. PDGF stimulates 32P incorporation preferentially into the Mr approximately 180,000 and less extensively into the Mr approximately 160,000 protein. EGF stimulates 32P incorporation predominantly into a protein of Mr approximately 160,000. The similarity of PDGF and EGF in stimulating phosphotyrosine-specific protein kinase activity and the stimulation of a similar activity by viral transformation (src) genes suggest that a common mechanism may exist for the phenotypic expression of increased DNA synthesis and cell growth stimulated by these separate factors.

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Selected References

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  1. Beemon K., Hunter T. Characterization of Rous sarcoma virus src gene products synthesized in vitro. J Virol. 1978 Nov;28(2):551–566. doi: 10.1128/jvi.28.2.551-566.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beemon K. Transforming proteins of some feline and avian sarcoma viruses are related structurally and functionally. Cell. 1981 Apr;24(1):145–153. doi: 10.1016/0092-8674(81)90510-9. [DOI] [PubMed] [Google Scholar]
  3. Carpenter G., King L., Jr, Cohen S. Epidermal growth factor stimulates phosphorylation in membrane preparations in vitro. Nature. 1978 Nov 23;276(5686):409–410. doi: 10.1038/276409a0. [DOI] [PubMed] [Google Scholar]
  4. Carpenter G., King L., Jr, Cohen S. Rapid enhancement of protein phosphorylation in A-431 cell membrane preparations by epidermal growth factor. J Biol Chem. 1979 Jun 10;254(11):4884–4891. [PubMed] [Google Scholar]
  5. Cheng Y. S., Chen L. B. Detection of phosphotyrosine-containing 34,000-dalton protein in the framework of cells transformed with Rous sarcoma virus. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2388–2392. doi: 10.1073/pnas.78.4.2388. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cohen S., Carpenter G., King L., Jr Epidermal growth factor-receptor-protein kinase interactions. Co-purification of receptor and epidermal growth factor-enhanced phosphorylation activity. J Biol Chem. 1980 May 25;255(10):4834–4842. [PubMed] [Google Scholar]
  7. Cohen S., Ushiro H., Stoscheck C., Chinkers M. A native 170,000 epidermal growth factor receptor-kinase complex from shed plasma membrane vesicles. J Biol Chem. 1982 Feb 10;257(3):1523–1531. [PubMed] [Google Scholar]
  8. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deuel T. F., Huang J. S., Proffitt R. T., Baenziger J. U., Chang D., Kennedy B. B. Human platelet-derived growth factor. Purification and resolution into two active protein fractions. J Biol Chem. 1981 Sep 10;256(17):8896–8899. [PubMed] [Google Scholar]
  10. Ehrhart J. C., Creuzet C., Rollet E., Loeb J. Epidermal growth factor-stimulated phosphorylation of tyrosine residues on a 120 000 dalton protein in mouse liver plasma membrane subfractions. Biochem Biophys Res Commun. 1981 Sep 30;102(2):602–609. doi: 10.1016/s0006-291x(81)80174-x. [DOI] [PubMed] [Google Scholar]
  11. Ek B., Westermark B., Wasteson A., Heldin C. H. Stimulation of tyrosine-specific phosphorylation by platelet-derived growth factor. Nature. 1982 Feb 4;295(5848):419–420. doi: 10.1038/295419a0. [DOI] [PubMed] [Google Scholar]
  12. Erikson E., Erikson R. L. Identification of a cellular protein substrate phosphorylated by the avian sarcoma virus-transforming gene product. Cell. 1980 Oct;21(3):829–836. doi: 10.1016/0092-8674(80)90446-8. [DOI] [PubMed] [Google Scholar]
  13. Feldman R. A., Hanafusa T., Hanafusa H. Characterization of protein kinase activity associated with the transforming gene product of Fujinami sarcoma virus. Cell. 1980 Dec;22(3):757–765. doi: 10.1016/0092-8674(80)90552-8. [DOI] [PubMed] [Google Scholar]
  14. Ghysdael J., Neil J. C., Vogt P. K. A third class of avian sarcoma viruses, defined by related transformation-specific proteins of Yamaguchi 73 and Esh sarcoma viruses. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2611–2615. doi: 10.1073/pnas.78.4.2611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ghysdael J., Neil J. C., Wallbank A. M., Vogt P. K. Esh avian sarcoma virus codes for a gag-linked transformation-specific protein with an associated protein kinase activity. Virology. 1981 Jun;111(2):386–400. doi: 10.1016/0042-6822(81)90342-1. [DOI] [PubMed] [Google Scholar]
  16. Hunter T., Cooper J. A. Epidermal growth factor induces rapid tyrosine phosphorylation of proteins in A431 human tumor cells. Cell. 1981 Jun;24(3):741–752. doi: 10.1016/0092-8674(81)90100-8. [DOI] [PubMed] [Google Scholar]
  17. Hunter T., Sefton B. M. Transforming gene product of Rous sarcoma virus phosphorylates tyrosine. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1311–1315. doi: 10.1073/pnas.77.3.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kawai S., Yoshida M., Segawa K., Sugiyama H., Ishizaki R., Toyoshima K. Characterization of Y73, an avian sarcoma virus: a unique transforming gene and its product, a phosphopolyprotein with protein kinase activity. Proc Natl Acad Sci U S A. 1980 Oct;77(10):6199–6203. doi: 10.1073/pnas.77.10.6199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. King L. E., Jr, Carpenter G., Cohen S. Characterization by electrophoresis of epidermal growth factor stimulated phosphorylation using A-431 membranes. Biochemistry. 1980 Apr 1;19(7):1524–1528. doi: 10.1021/bi00548a040. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  22. Levinson A. D., Courtneidge S. A., Bishop J. M. Structural and functional domains of the Rous sarcoma virus transforming protein (pp60src). Proc Natl Acad Sci U S A. 1981 Mar;78(3):1624–1628. doi: 10.1073/pnas.78.3.1624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Levinson A. D., Oppermann H., Levintow L., Varmus H. E., Bishop J. M. Evidence that the transforming gene of avian sarcoma virus encodes a protein kinase associated with a phosphoprotein. Cell. 1978 Oct;15(2):561–572. doi: 10.1016/0092-8674(78)90024-7. [DOI] [PubMed] [Google Scholar]
  24. MITCHELL H. K., LUNAN K. D. TYROSINE-O-PHOSPHATE IN DROSOPHILA. Arch Biochem Biophys. 1964 Jul 20;106:219–222. doi: 10.1016/0003-9861(64)90179-1. [DOI] [PubMed] [Google Scholar]
  25. Neil J. C., Ghysdael J., Vogt P. K. Tyrosine-specific protein kinase activity associated with p105 of avian sarcoma virus PRCII. Virology. 1981 Feb;109(1):223–228. doi: 10.1016/0042-6822(81)90493-1. [DOI] [PubMed] [Google Scholar]
  26. Nishimura J., Deuel T. F. Stimulation of protein phosphorylation in Swiss mouse 3T3 cells by human platelet derived growth factor. Biochem Biophys Res Commun. 1981 Nov 16;103(1):355–361. doi: 10.1016/0006-291x(81)91700-9. [DOI] [PubMed] [Google Scholar]
  27. Plimmer R. H. Esters of phosphoric acid: Phosphoryl hydroxyamino-acids. Biochem J. 1941 Apr;35(4):461–469. doi: 10.1042/bj0350461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sefton B. M., Hunter T., Ball E. H., Singer S. J. Vinculin: a cytoskeletal target of the transforming protein of Rous sarcoma virus. Cell. 1981 Apr;24(1):165–174. doi: 10.1016/0092-8674(81)90512-2. [DOI] [PubMed] [Google Scholar]
  29. Ushiro H., Cohen S. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J Biol Chem. 1980 Sep 25;255(18):8363–8365. [PubMed] [Google Scholar]
  30. Whittenberger B., Glaser L. Inhibition of DNA synthesis in cultures of 3T3 cells by isolated surface membranes. Proc Natl Acad Sci U S A. 1977 Jun;74(6):2251–2255. doi: 10.1073/pnas.74.6.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]

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