Skip to main content
SpringerLink
Log in

Epidermal growth factor receptor-protein kinase interactions in hepatic membranes of rainbow trout (Oncorhynchus mykiss)

Fish Physiology and Biochemistry volume 22, pages 181–189 (2000)Cite this article

Abstract

The association between the presence of EGF binding and its associated tyrosine kinase activity was investigated in the hepatic membranes of rainbow trout (Oncorhynchus mykiss). Time course experiments indicted that the binding of EGF to hepatic membranes was temperature dependent and resulted in increased phosphorylation of membrane components. The phosphorylation reaction was dependent on the presence of divalent ions (Mg2+ and Mn2+) but not monovalent ions (Na+ and K+) or calcium. Western blot analysis of phosphorylated trout membranes indicated protein bands at 150 and 180 kDa which is similar to that observed in mammals. Use of specific substrates and protein kinase inhibitors also indicated that the increase in EGF stimulated phosphorylation was related to an increase in tyrosine kinase activity. These data support the hypothesis that trout have an EGF-like receptor that has binding and biological activities similar to that observed in mammalian species.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

USD 39.95

Price includes VAT (Canada)

References

  • Carpenter, G. 1987. Receptors for epidermal growth factor and other polypeptide mitogens. Ann. Rev. Biochem. 56: 881-914.

    Google Scholar 

  • Carpenter, G. and Cohen, S. 1979. Epidermal growth factor. Ann. Rev. Biochem. 48: 193-216.

    Google Scholar 

  • Carpenter, G., King, L., Jr. and Cohen, S. 1979. Rapid enhancement of protein phosphorylation in A-431 cell membrane preparations by epidermal growth factor. J. Biol. Chem. 254: 4884-4891.

    Google Scholar 

  • Cohen, S. 1983. The epidermal growth factor (EGF). Cancer 51: 1787-1791.

    Google Scholar 

  • Cohen, S., Carpenter, G.A. and King, L., Jr. 1980. Epidermal growth factor-receptor-protein kinase interactions: co-purification of receptor and epidermal growth factor-enhanced phosphorylation. J. Biol. Chem. 255: 4834-4842.

    Google Scholar 

  • Gazit, A., Yaish, P., Gilon, C. and Levitzki, A. 1989. Tyrphostins 1: Synthesis and biological activity of protein tyrosine kinase inhibitors. J. Med. Chem. 32: 2344-2352.

    Google Scholar 

  • Gospodarowicz, D. 1981. Epidermal and nerve growth factors in mammalian development. Ann. Rev. Physiol. 43: 251-263.

    Google Scholar 

  • Gregoriou, M. and Rees, A. R. 1984. Properties of monoclonal antibody to epidermal growth factor receptor with implications for the mechanism of action of EGF. The EMBO J. 3: 929-937.

    Google Scholar 

  • Hunter, T. and Cooper, I.A. 1985. Protein tyrosine kinases. Ann. Rev. Biochem. 54: 897-930.

    Google Scholar 

  • Klein, H.H., Freidenberg, G.R., Cordera, R. and Olefsky, J.M. 1985. Substrate specificities of insulin and epidermal growth factor receptor kinases. Biochem. Biophys. Res. Comm. 127: 254-263.

    Google Scholar 

  • Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, Lond. 227: 680-685.

    Google Scholar 

  • Lowry, O.H., Rosenbrough, D.D., Farr, A.L.I. and Randall, R.J. 1951. Protein measurement with the folin reagent. J. Biol. Chem. 193: 265-275.

    Google Scholar 

  • Matter, W.F., Brown, R.F., and Vlahos, C.J. 1992. The inhibition of phosphatdylinositol 3-kinase by quercetin and analogs. Biochem. Biophys. Res. Comm. 186: 624-631.

    Google Scholar 

  • Malitschek, B., Wittbrodt, J., Fischer, P., Lammers, R., Ulrich, A. and Schartl, M. 1994. Autocrine stimulation of Xmrk receptor tyrosine kinase in Xiphophorus melanomacells and identification of a source for the physiological ligand. J. Biol. Chem. 269: 10423-10430.

    Google Scholar 

  • Margolis, B., Lax, I., Kris, R., Dombalagian, M., Honegger, A.M., Hocuk, R., Givol, D., Ullkrich, A. and Schlessinger, J. 1989. All autophosphorylation sites of epidermal growth factor (EGF) receptor and BER2/neu are located in their carboxy-terminal tails: identification of a novel site in EGF receptors. J. Biol. Chem. 264: 10667-10671.

    Google Scholar 

  • Moolenaar, W.H., Bierman, A.J., Tilly, B.C., Verlaan, I., Honegger, A.M., Ullrich, A. and Schlessinger, J. 1988. A point mutation at the ATP-binding site of the EGF-receptor abolishes signal transduction. ENMO J. 7: 707-710.

    Google Scholar 

  • Newsted, J. and Gesy, J. 1991. Characterization of epidermal growth factor binding to the hepatic plasma membrane of rainbow trout (Oncorhynchus mykiss). Gen. Comp. Endocrinol. 83: 345-353.

    Google Scholar 

  • Pike, L. 1983. Assay of growth factor stimulated tyrosine kinases using synthetic peptide substrates. InMethods in Enzymology. Vol. 146, Peptide Growth Factors. pp. 353-362. Edited by D. Barnes and D.A. Sirbasku. Academic Press, New York.

    Google Scholar 

  • Ritvos, O., Jalkanen, J., Pekonen, F., Stenman, U.M. and Ranta, T. 1988. Epidermal growth factor but not insulin-like growth fact potentiates adenosine 3',5'-monophosphate-mediated chorionic gonadotropin secretion by cultures human choriocarcinoma cells. Endocrinology 123: 859-865.

    Google Scholar 

  • Schlessinger, J. 1988. The epidermal growth factor receptor as a multifunctional allosteric protein. Biochemistry 27: 3119-3123.

    Google Scholar 

  • Schonbrunn, A., Kranshoff, M., Westendorf, J.M. and Tashjian, A.H., Jr. 1980. Epidermal growth factor and thyrotropinreleasing hormone act similarly on a clonal pituitary cell strain. J. Cell Biol. 85: 786-797.

    Google Scholar 

  • Shibasaki, F., Fukui, Y. and Takenawa, T. 1993. Different properties of monomer and heterodimer forms of phosphatidylinositol 3-kinase. Biochem. J. 289: 227-231.

    Google Scholar 

  • Shiraishi, T., Domoto, T., Imai, N., Shimada, Y. and Watanabe, K. 1987. Specific inhibitors of tyrosine-specific protein kinase, synthetic 4-hydroxycinnamamide derivatives. Biochem. Biophys. Res. Comm. 147: 322-328.

    Google Scholar 

  • Steel, R. G. and Torrie, J.H. 1980. Principles and Procedures in Statistics. 2nd Edition, McGraw-Hill, New York.

    Google Scholar 

  • Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins form polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350-4354.

    Google Scholar 

  • Ushiro, H. and Cohen, S. 1986. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J. Biol. Chem. 255: 8363-8365.

    Google Scholar 

  • Velu, T.J. 1990. Structure, function and transforming potential of the epidermal growth factor receptor. Mol. Cell. Endocrinol. 7: 205-216.

    Google Scholar 

  • Zechel, C., Peters, H., Schleenbecker, U. and Anders, F. 1992. Expression of genes related to the human erbb, erba, pdgf and pdgf-r in tumors of different etiology in Xiphophorus. Int. J. Cancer 52: 66-75.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Forestty and Wildlife Management, University of Massachusetts at Amherst, 217 Holdsworth Natural Resources Center, Amherst, MA, 01003, USA

    J.L. Newsted

  2. Department of Zoology, Natural Science Building, Michigan State University, USA

    J.P. Giesy

Authors
  1. J.L. Newsted
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J.P. Giesy
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Newsted, J., Giesy, J. Epidermal growth factor receptor-protein kinase interactions in hepatic membranes of rainbow trout (Oncorhynchus mykiss). Fish Physiology and Biochemistry 22, 181–189 (2000). https://doi.org/10.1023/A:1007872209650

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1007872209650

  • fish EGF
  • receptor
  • tyrosine kinase
  • western blot

Search

Navigation