Designing biosensors for Rho family proteins--deciphering the dynamics of Rho family GTPase activation in living cells.

Rho family GTPases are molecular switches that couple changes in the extracellular environment to intracellular signal transduction pathways. Their ability to regulate behaviors such as cell motility suggests very tight kinetic and spatial control of their activity, which is missed in most biochemical assays. Fluorescent probes that non-invasively report the changing subcellular location of Rho GTPase activity in single living cells are now allowing us to examine spatio-temporal regulation of the activity of these proteins, and are providing new biological insights. Several strategies can be used to construct such probes, and there are advantages and disadvantages associated with the diverse probe designs.