Abstract

Tyrphostins are synthetic compounds that have been described as in vitro and in vivo inhibitors of epidermal growth factor receptor tyrosine kinase activity. In NIH/3T3 cells transfected with the c-src/F527 gene, an increase in the level of tyrosine phosphorylation of several proteins, including pp125FAK, within a group of proteins of 120 kDa, of p85 (cortactin), and of p62 is observed, which is due to the elevated kinase activity of the resulting encoded pp60F527 protein. In the transfected cells, we showed that the tyrphostins we used, i.e., AG18, AG34, and AG82, strongly diminished the tyrosine phosphorylation of these proteins. Analysis of the steady state level of pp60F527 in tyr-phostin-treated cells revealed that AG34 and AG82, the two most potent compounds, also induced 30 and 48% decreases, respectively, in the amount of pp60F527, while having no action on the levels of other proteins, especially the pp60F527 kinase substrates. Measurement of the rates of pp60F527 synthesis and degradation showed that this decreased level was due to a slower rate of synthesis in the presence of AG34 and AG82. Tyrphostins also reversed the pp60F527-induced transformed morphology of NIH/3T3 cells and also inhibited the pp60F527 kinase activity in vitro. We conclude that the effects elicited by the tyrphostins occurred not only through the inhibition of the pp60F527 protein kinase activity but also through a selective reduction of the Src protein steady state level in the cases of AG34 and AG82. This is a novel mode of action for these two tyrphostins, which were the most active compounds in this system.