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PMID: 20235099 Published · ppublish English Journal Article

Biosensors for characterizing the dynamics of rho family GTPases in living cells.

Current protocols in cell biology ·Vol. Chapter 14 ·2010-03-00 ·Pages Unit 14.11.1-26

Hodgson L, Shen F, Hahn K

Abstract

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.

MeSH Terms
Animals Biosensing Techniques/methods Cell Survival Cells/cytology,enzymology Fluorescent Dyes/metabolism Green Fluorescent Proteins/metabolism Image Processing, Computer-Assisted Mice Molecular Imaging cdc42 GTP-Binding Protein/metabolism rac1 GTP-Binding Protein/metabolism rho GTP-Binding Proteins/metabolism
Chemicals
Fluorescent Dyes enhanced green fluorescent protein Green Fluorescent Proteins cdc42 GTP-Binding Protein rac1 GTP-Binding Protein rho GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hodgson Louis
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine of Yeshiva University, Gruss-Lipper Biophotonics Center, Bronx, New York, USA.
Shen Feimo
Hahn Klaus
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Article Info
Journal
Current protocols in cell biology
Abbr.
Curr Protoc Cell Biol
ISSN
1934-2616
DOI
10.1002/0471143030.cb1411s46
Published
2010-03-00
Pages
Unit 14.11.1-26
Language
English
Region
United States
NLM ID
101287856
PMCID
PMC2998069
Subset
IM
Grants
NIGMS NIH HHS · R01 GM057464 · United States
NIGMS NIH HHS · R01 GM057464-10 · United States
External Links
PubMed source DOI: 10.1002/0471143030.cb1411s46 PMC Full Text
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