Epidermal Growth Factor Mediated Signal Transduction in Rat Hepatocytes

  • Lijun Yang
  • György Baffy
  • Sue Goo Rhee
  • John R. Williamson
Conference paper
Part of the NATO ASI Series book series (volume 52)

Summary

Treatment of rat hepatocytes with epidermal growth factor (EGF) produced an enhanced tyrosine phosphorylation of the EGF receptor and phospholipase C-γ (PLC-γ) in conjunction with the mobilization of Ca2+. Approximately 30% of the total PLC-γ was tyrosine phosphorylated with a maximum being reached after 30s of incubation with EGF. Pretreatment of the rats with pertussis toxin prior to isolation of the hepatocytes blocked EGF-induced tyrosine phosphorylation of PLC-γ and Ca2+ mobilization but had no effect on autophosphorylation of the EGF receptor or Ca2+ responses elicited by angiotensin II or phenylephrine. Under these conditions Gi-protein α subunits were fully ADP-ribosylated. Antibodies specific to PLC-γ coimmunoprecipitated phosphorylated proteins with molecular masses of 190kDa, 96kDa, 47kDa and 25kDa but not the EGF receptor. A 41kDa Gi-protein α subunit was found to be present in the anti-PLC-γ immune complex as shown by in vitro ADP-ribosylation using [32P]NAD+ and activated pertussis toxin. Antibodies specific for the EGF receptor also coimmunoprecipitated a Gi-protein α subunit. Treatment of hepatocytes with EGF caused a decrease in the amount of Gi-protein α subunit associated with the EGF receptor and an increase in its association with PLC-γ. These results indicate that a pertussis toxin-sensitive Gi-protein is intimately involved in the signal transduction pathway mediating EGF-induced activation of PLC-γ and Ca2+ mobilization in hepatocytes.

Keywords

Corn Tyrosine Serine Angiotensin Hepes 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1991

Authors and Affiliations

  • Lijun Yang
    • 1
  • György Baffy
    • 1
  • Sue Goo Rhee
    • 2
  • John R. Williamson
    • 1
  1. 1.Dept. of Biochemistry and BiophysicsUniversity of PennsylvaniaPhiladelphiaUSA
  2. 2.Laboratory of BiochemistryNHLBI Institute NIHBethesdaUSA

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