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

Abstract

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 GloTM 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.

Footnotes

* The revised version includes an additional supplement (S11).

* http://doi.org/10.4121/ac196f25-1c20-4c0c-a0b9-f01cd3fadc45