Decreased binding and autophosphorylation of the epidermal growth factor receptor in ethanol-fed rats.
We have shown previously that binding and processing of epidermal growth factor are impaired in livers of ethanol-fed rats. In the current study, we examined these ethanol-induced alterations in greater detail by studying both high and low affinity epidermal growth factor binding as well as the ability of added ligand to stimulate receptor autophosphorylation. We also measured the binding of anti-receptor antibody to intact and permeabilized cells in order to determine more accurately the levels of receptor protein. Hepatocytes were isolated from ethanol-fed and pair-fed control rats. Ligand binding, antibody binding, and ligand-induced receptor autrophosphorylation were measured in the respective cell populations. In ethanol-fed animals, binding to both high and low affinity states of the hepatic epidermal growth factor receptor was decreased by 40-50% (P < 0.01). This ethanol-induced decrease in ligand binding was accompanied by a reduced ability of epidermal growth factor to stimulate receptor autophosphorylation (32% decrease, P < 0.01). In contrast, binding of anti-receptor antibody was not altered in ethanol-fed animals. In conclusion, chronic ethanol feeding decreased epidermal growth factor binding with a concomitant decrease in the ability of the receptor tyrosine kinase to phosphorylate tyrosine residues. These changes were not accompanied by an actual decrease in receptor protein content. These findings could be relevant to modified responses to this growth factor in the livers of chronic ethanol-fed animals.
O'Rourke
M F
MF
Department of Internal Medicine, University of Nebraska Medical Center, Omaha, U.S.A.
Tuma
D J
DJ
Casey
C A
CA
D000818:Animals; D015153:Blotting, Western; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000187:drug effects / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D010766:Phosphorylation; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D000431:Ethanol; D004815:Epidermal Growth Factor; D066246:ErbB Receptors