Open AccessBiosensors for Characterizing the Dynamics of Rho Family GTPases in Living Cells
Author(s) -
Hodgson Louis,
Shen Feimo,
Hahn Klaus
Publication year - 2010
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/0471143030.cb1411s46
Subject(s) - biosensor , förster resonance energy transfer , cdc42 , gtpase , nanotechnology , intramolecular force , chemistry , fluorescence , materials science , biochemistry , physics , stereochemistry , quantum mechanics
The biosensors developed in the authors' laboratory have been based on different designs, each imparting specific strengths and weaknesses. Here we describe detailed protocols for the application of three biosensors exemplifying different designs—first, a design in which an environmentally sensitive dye is used to report the activation of endogenous Cdc42, followed by two biosensors based on FRET, one using intramolecular and the other intermolecular FRET. The design differences lead to the need for different approaches in imaging and image analysis. Curr. Protoc. Cell Biol . 46:14.11.1‐14.11.26. © 2010 by John Wiley & Sons, Inc.