Use of an anti-insulin receptor antibody to discriminate between metabolic and mitogenic effects of insulin: correlation with receptor autophosphorylation.

In a previous report we described the properties of a rabbit anti-insulin receptor antibody (RAIR-IgG) and its effects on the autophosphorylation and kinase activity of human insulin receptors. The present study was carried out on the hepatoma cell line Fao. We tested the mimetic effects of RAIR-IgG on different biological parameters known to be stimulated by insulin, receptor autophosphorylation and kinase activity. RAIR-IgG stimulated the metabolic effects (glucose and amino acid transport) but, unlike insulin, was unable to promote cell proliferation. These data clearly demonstrated the existence of two distinctly controlled pathways in the mediation of the hormonal response. When we investigated the effects of this antibody at the molecular level we found that in a cell-free system RAIR-IgG weakly stimulated receptor autophosphorylation on non-regulatory sites and failed to stimulate tyrosine kinase activity toward exogenous substrates. Accordingly, RAIR-IgG did not stimulate receptor autophosphorylation in 32P-labelled intact cells. Interestingly, under similar conditions RAIR-IgG elicited ribosomal S6 protein phosphorylation, as did insulin. The possibility that RAIR-IgG activated a cryptic tyrosine kinase activity is discussed.