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

Abstract

AbstractIn 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.