Related Experiment Video
Updated: Sep 10, 2025

Comparing the Affinity of GTPase-binding Proteins using Competition Assays
Published on: October 8, 2015
Split-Small GTPase Reassembly as a Method to Control Cellular Signaling with User-Defined Inputs
Yuchen He1, Benjamin M Faulkner1, Rachel S Weatherford1
1Department of Chemistry, University of Virginia, Charlottesville, Virginia 22904, United States.
Abstract:
Small GTPases are critical signaling enzymes that control diverse cellular functions, such as cell migration and proliferation. However, dissecting the roles of these enzymes in cellular signaling is hindered by the lack of a plug-and-play methodology for the direct, temporal control of small GTPase activity by using user-defined inputs. Herein, we present a method that pairs split-small GTPases with user-defined chemical inducer of dimerization (CID) systems in a plug-and-play manner to directly control small GTPase signaling in living cells. The modularity of split-small GTPase systems allows for the selection of CIDs with minimal off-target effects on the pathway being studied. Our results highlight the ability to obtain consistent pathway activation with varying CID systems for direct control of MAPK signaling, filopodia formation, and cell retraction. Thus, split-small GTPase systems provide a customizable platform for the development of temporally gated systems for directly controlling cellular signaling with user-defined inputs.
More Related Videos
10:27Spatio-Temporal Manipulation of Small GTPase Activity at Subcellular Level and on Timescale of Seconds in Living Cells
Published on: March 9, 2012
13:51Detection of Small GTPase Prenylation and GTP Binding Using Membrane Fractionation and GTPase-linked Immunosorbent Assay
Published on: November 11, 2018
Related Concept Videos
GTPases and their Regulation
Large G-proteins,...
Small GTPases - Ras and Rho
Three regulatory proteins control their activity:
Activation and Inactivation of G Proteins
Amplifying Signals via Enzymatic Cascade
Assembly of Signaling Complexes
Interaction domains in cell signaling
Interaction domains recognize exposed features of their binding partners containing post-translationally modified sequences,...
Rab Cascades