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PMID: 3011811 Published · ppublish English Journal Article

Early events elicited by bombesin and structurally related peptides in quiescent Swiss 3T3 cells. I. Activation of protein kinase C and inhibition of epidermal growth factor binding.

The Journal of cell biology ·Vol. 102 ·No. 6 ·1986-06-00 ·Pages 2211-22

Zachary I, Sinnett-Smith JW, Rozengurt E

Abstract

Addition of bombesin to quiescent cultures of Swiss 3T3 cells caused a rapid increase in the phosphorylation of an Mr 80,000 cellular protein (designated 80k). The effect was both concentration and time dependent; enhancement in 80k phosphorylation could be detected as early as 10 s after the addition of peptide. Recently, a rapid increase in the phosphorylation of an 80k cellular protein after treatment with phorbol esters or diacylglycerol has been shown to reflect the activation of protein kinase C in intact fibroblasts (Rozengurt, E., A. Rodriguez-Pena, and K. A. Smith, 1983, Proc. Natl. Acad. Sci. USA., 80:7244-7248; Rozengurt, E., A. Rodriguez-Pena, M. Coombs, and J. Sinnett-Smith, 1984, Proc. Natl. Acad. Sci. USA., 81:5748-5752). The 80k phosphoproteins generated in response to bombesin and to phorbol 12,13-dibutyrate were identical as judged by one- and two-dimensional PAGE and by peptide mapping after partial proteolysis with Staphylococcus aureus V8 protease. In addition, prolonged pretreatment of 3T3 cells with phorbol 12,13-dibutyrate, which leads to the disappearance of protein kinase C activity, blocked the ability of bombesin to stimulate 80k. Bombesin also caused a rapid (1 min) inhibition of 125I-labeled epidermal growth factor (125I-EGF) binding to Swiss 3T3 cells. The inhibition was both concentration and temperature dependent and resulted from a marked decrease in the affinity of the EGF receptor for its ligand. Peptides structurally related to bombesin, including gastrin-releasing peptide, also stimulated 80k phosphorylation and inhibited 125I-EGF binding; both effects were selectively blocked by a novel bombesin antagonist. These results strongly suggest that these responses are mediated by specific high-affinity receptors that recognize the peptides of the bombesin family in Swiss 3T3 cells. While an increase in cytosolic Ca2+ concentration does not mediate the bombesin inhibition of 125I-EGF binding, the activation of protein kinase C in intact Swiss 3T3 cells by peptides of the bombesin family may lead to rapid inhibition of the binding of 125I-EGF to its cellular receptor.

MeSH Terms
Animals Binding, Competitive/drug effects Bombesin/metabolism,pharmacology Calcium/physiology Cell Line Cytosol/physiology Enzyme Activation/drug effects Epidermal Growth Factor/metabolism ErbB Receptors Interphase/drug effects Mice Molecular Weight Phorbol 12,13-Dibutyrate Phorbol Esters/pharmacology Phosphoproteins/metabolism Phosphorylation Protein Kinase C/metabolism,physiology Receptors, Bombesin Receptors, Cell Surface/drug effects,metabolism,physiology Temperature
Chemicals
Phorbol Esters Phosphoproteins Receptors, Bombesin Receptors, Cell Surface Phorbol 12,13-Dibutyrate Epidermal Growth Factor ErbB Receptors Protein Kinase C Bombesin Calcium
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Zachary I
Sinnett-Smith J W
Rozengurt E
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72 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.102.6.2211
Published
1986-06-00
Pages
2211-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2114266
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.102.6.2211 PMC Full Text View journal details
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