MONTE-CARLO SIMULATIONS OF MEMBRANE SIGNAL-TRANSDUCTION EVENTS - EFFECT OF RECEPTOR BLOCKERS ON G-PROTEIN ACTIVATION

Cells have evolved elaborate strategies for sensing, responding to, and interacting with their environment. In many systems, interaction of cell surface receptors with extracellular ligand can activate cellular signal transduction pathways leading to G-protein activation and calcium mobilization. In BC(3)H1 smooth muscle-like cells, we find that the speed of calcium mobilization as well as the fraction of cells which mobilize calcium following phenylephrine stimulation is dependent upon receptor occupation. To determine whether receptor inactivation affects calcium mobilization, we use the receptor antagonist prazosin to block a fraction of cell surface receptors prior to phenylephrine stimulation. For cases of equal receptor occupation by agonist, cells with inactivated or blocked receptors show diminished calcium mobilization following phenylephrine stimulation as compared to cells without inactivated receptors. Ligand/receptor binding and two-dimensional diffusion of receptors and G-proteins in the cell membrane are studied using a Monte Carlo model. The model is used to determine if receptor inactivation affects G-protein activation and thus the following signaling events for cases of equal equilibrium receptor occupation by agonist. The model predicts that receptor inactivation by antagonist binding results in lower G-protein activation not only by reducing the number of receptors able to bind agonist but also by restricting the movement of agonist among free receptors. The latter process is important to increasing the access of bound receptors to G-proteins.