Techniques used for functional characterization of polyamine transporters

Abstract

Transport systems are key processes in every living organism: they allow the entry of all essential nutrients into the cell and its compartments and regulate the intracellular concentrations of metabolites. The transport of cell nutrients represents the first step of many metabolic routes and may also regulate such processes. They are also responsible for reaching the effective intracellular con-centration of therapeutic drugs and some mechanisms of resistance and tolerance also depend on them. However, the common techniques used to evaluate the me-tabolites transport in different cells types are not easy to carry out and require an extensive training. In this chapter we made detailed protocols and tips about the expression of transporters, different activity assays and transporter kinetics deter-mination.