Signal transmission by epidermal growth factor receptor: coincidence of activation and dimerization

Dimerization of epidermal growth factor receptor dissolved in a solution of nonionic detergent was followed with a resolution of 1 min by quantitative cross-linking with glutaraldehyde. Upon addition of epidermal growth factor to the solution, the initially monomeric protein dimerized in a reaction that was second-order in the concentration of receptor. A second-order rate constant, on the basis of enzymatic activity as a measure of the concentration of functional receptor, was calculated from time courses of dimerization at various initial concentrations of receptor. The activation of the protein tyrosine kinase of the receptor was monitored directly under the same conditions with an exogenous substrate. The increase in tyrosine kinase activity displayed kinetics that were also second-order in the concentration of receptor. A second-order rate constant for the activation of the tyrosine kinase could be calculated from the time courses. The second-order rate constant for the activation of the tyrosine kinase by epidermal growth factor was indistinguishable from the second-order rate constant for the dimerization induced by epidermal growth factor. Therefore, dimerization of epidermal growth factor receptor and activation of its tyrosine kinase are coincident events, both initiated by the binding of epidermal growth factor.