Quantification of GTPase cycling rates of GTPases and GTPase : effector mixtures using GTPase Glo™ assays

In different cellular activities like signal transduction, cell division, and intracellular transportation, small GTPases take on a vital role. Their functioning involves hydrolysing guanosine triphosphate (GTP) to guanosine diphosphate (GDP). In this article we explain the application of a commercially accessible GTPase assay, known as the GTPase Glo assay by Promega, for the quantitative investigation of GTPase - effector interactions and the interplay between effectors. Basic ProtocolConducting GTPase assays with GTPase : effector protein mixtures using the GTPase Glo assay (Promega). Supporting Protocol 1Analysing GTPase assays to correlate the assay readout (luminescence) to amount of remaining GTP. Supporting Protocol 2Fitting GTPase assay data to obtain GTPase cycling rates.