Cell
Volume 25, Issue 2, August 1981, Pages 433-440
Journal home page for Cell

Article
A steady state model for analyzing the cellular binding, internalization and degradation of polypeptide ligands

https://doi.org/10.1016/0092-8674(81)90061-1Get rights and content

Abstract

We demonstrate that the interaction of polypeptide ligands with cells under physiological conditions can be described by a set of steady state equations. These equations include four new rate constants: Vr, the rate of insertion of receptors into the cell membrane; Ke, the endocytotic rate constant of occupied receptors; Kt, the turnover rate constant of unoccupied receptors; and Kh, the rate constant of hydrolysis of internalized ligand. Several simple procedures are described for determining these constants. In experiments in which epidermal growth factor and human fibroblasts were used, the cell-ligand interactions followed the predictions of the steady state model. The utility of the steady state equations is demonstrated by establishing the kinetic basis of the commonly observed “down regulation” phenomenon and by quantitating the effect of methylamine on the endocytotic and degradation rates of epidermal growth factor. We also show that the slope of a “Scatchard plot” of steady state binding data is a complex constant including terms for the endocytotic rate of both occupied and unoccupied receptors. The X-intercept of such a plot is a function of the insertion rate of new receptors, the internalization rate of occupied receptors and the degradation rate of the internalized ligand. The steady state equations allow one to predict changes in cellular ligand binding resulting from alterations in the four rate constants. They also provide a foundation for computer simulations of ligand-cell interactions, which closely correspond to experimental data. These approaches should facilitate studies on the control of cellular activities by these polypeptide ligands.

References (30)

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