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 Protocol Conducting GTPase assays with GTPase : effector protein mixtures using the GTPase Glo™ assay (Promega). Supporting Protocol 1 Analysing GTPase assays to correlate the assay readout (luminescence) to amount of remaining GTP. Supporting Protocol 2 Fitting GTPase assay data to obtain GTPase cycling rates. ### Competing Interest Statement The authors have declared no competing interest.