Analysis of intracellular receptor/ligand sorting. Calculation of mean surface and bulk diffusion times within a...

Cell surface receptors bind extracellular ligand molecules and transport those ligands into the cell by a process termed receptor-mediated endocytosis. Receptor and ligand molecules are sorted from one another after endocytosis, apparently within a structure consisting of intracellular vesicles and connected thin tubules. The experimental observation is that most free (unbound) ligand molecules are found in the lumen of the vesicles and receptors are located primarily within the tubules. Because equilibrium and geometric considerations do not explain this segregation, a kinetic scheme involving the passive diffusion of molecules from a vesicle into a tubule is investigated. Two possible sorting mechanisms are considered: first, that receptors are able to move into tubules more rapidly than ligand molecules due to an advantage in dimensionality and, second, that receptors diffusing into tubules are trapped there while ligands are not. Mean diffusion times for receptor and ligand movement into a tubule are calculated by solving Poisson's equation in two and three dimensions, respectively, on the surface of and within a sphere. Using estimated parameter values, we found that only the second scheme is able to account for the experimentally observed sorting. An estimate is obtained for the length of time a tubule and vesicle must be connected in order to remove a significant number of receptors into a tubule. The fraction of free ligand that is "mis-sorted" with the recycling receptor population and thus exocytosed is also determined.