Quantitative Analysis of Integrin Trafficking.

The last two decades of research into integrin trafficking has revealed fascinating insight into the function of integrin receptors, particularly in the context of cell invasion and migration in cancer. Deregulation in the trafficking pathways of integrin receptors contributes to a variety of pathological conditions including cancer, and in fact, altered endocytic trafficking of these receptors has been shown to drive transformation and tumor progression. Being able to experimentally measure integrin internalization, recycling and cell surface levels are vital for determining the role integrins play in health and disease. Surface-labeling based endocytic trafficking assays provide a way to experimentally measure changes in the rate of internalization of cell surface proteins, and the recycling of internalized proteins back to the cell surface, with high accuracy. This chapter will focus on quantitative approaches based on cell surface biotinylation and capture ELISA to measure endocytosis, recycling, and cell surface levels of integrin receptors.