A screening-compatible live cell fluorescence resonance energy transfer-based assay for modulation of Rho GTPase activity.

rho GTP-Binding Proteins / metabolism Fluorescence Resonance Energy Transfer Rho Guanine Nucleotide Exchange Factors / metabolism GTPase-Activating Proteins / metabolism
Chemistry Energy Environmental sciences Material sciences Physics

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
16 12 2022
Historique:
received: 27 05 2022
revised: 08 07 2022
accepted: 23 08 2022
pubmed: 24 9 2022
medline: 22 12 2022
entrez: 23 9 2022
Statut: ppublish

Résumé

Rho family GTPases are central regulators of cytoskeletal dynamics controlled by guanine nucleotide exchange factors (RhoGEFs) and GTPase-activating proteins (RhoGAPs). This protocol presents a workflow for a robust high-throughput compatible biosensor assay to analyze changes in Rho GTPase activity by these proteins in the native cellular environment. The procedure can be used for semi-quantitative comparison of GEF/GAP function and extended for analysis of additional modulators. The experimental design is applicable also to other monomolecular ratiometric FRET sensors. For complete details on the use and execution of this protocol, please refer to Müller et al. (2020).

Identifiants

DOI: 10.1016/j.xpro.2022.101705 PMID: 36149795 PMC: PMC9508600
pubmed: 36149795
pii: S2666-1667(22)00585-8
doi: 10.1016/j.xpro.2022.101705
pmc: PMC9508600
pii:
doi:

Substances chimiques

rho GTP-Binding Proteins EC 3.6.5.2
Rho Guanine Nucleotide Exchange Factors 0
GTPase-Activating Proteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

101705

Informations de copyright

Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

Auteurs

Paul Markus Müller (PM)

Institute of Chemistry and Biochemistry, Freie Universität Berlin, Thielallee 63, 14195 Berlin, Germany. Electronic address: paulmarkus.mueller@fu-berlin.de.

Oliver Rocks (O)

Institute of Biochemistry, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. Electronic address: oliver.rocks@charite.de.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides de signalisation intracellulaire › Protéines G › Protéines G monomériques › Protéines G rho › A screening-compatible live cell fluorescence...
questionsmedicales.fr › Phénomènes chimiques › Phénomènes biochimiques › Transfert d'énergie par résonance de fluorescence › A screening-compatible live cell fluorescence...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides de signalisation intracellulaire › Régulateurs des protéines G › Facteurs d'échange de nucléotides guanyliques › Rho guanine nucleotide exchange factors › A screening-compatible live cell fluorescence...
questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines et peptides de signalisation intracellulaire › Régulateurs des protéines G › Protéines d'activation de la GTPase › A screening-compatible live cell fluorescence...