ArticleBMC biology2011
Real-time visualization of heterotrimeric G protein Gq activation in living cells.
Article in BMC biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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Who cites it
47 citing papers in PubMed, 96 citations in OpenAlex.
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- Direct Activity Measurement of Heterotrimeric Gi Proteins and Gq Protein By Effector Pulldown.Bio-protocol · 2025Article
- Tissue-specific consequences of tag fusions on protein expression in transgenic mice.PLoS genetics · 2025Article
- Get Ready to Sharpen Your Tools: A Short Guide to Heterotrimeric G Protein Activity Biosensors.Molecular pharmacology · 2024Review
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- Advancements in G protein-coupled receptor biosensors to study GPCR-G protein coupling.British journal of pharmacology · 2023Review
- Mapping the conformational landscape of the stimulatory heterotrimeric G protein.Nature structural & molecular biology · 2023Article
- Genetically encoded tools for in vivo G-protein-coupled receptor agonist detection at cellular resolution.Clinical and translational medicine · 2022Review
- Gq Signaling in Autophagy Control: Between Chemical and Mechanical Cues.Antioxidants (Basel, Switzerland) · 2022Review
- Flow cytometry based-FRET: basics, novel developments and future perspectives.Cellular and molecular life sciences : CMLS · 2022Review
- BRET-Based Biosensors to Measure Agonist Efficacies in Histamine HInternational journal of molecular sciences · 2022Article
- Designer proteins that competitively inhibit GαThe Journal of biological chemistry · 2021Article
- Two Motors and One Spring: Hypothetic Roles of Non-Muscle Myosin II and Submembrane Actin-Based Cytoskeleton in Cell Volume Sensing.International journal of molecular sciences · 2021Review
- Imaging of Genetically Encoded FRET-Based Biosensors to Detect GPCR Activity.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Multiple GPCR Functional Assays Based on Resonance Energy Transfer Sensors.Frontiers in cell and developmental biology · 2021Review
- Pharmacological Characterization of the Stick InsectACS omega · 2020Article
- Kinetic Analysis of the Early Signaling Steps of the Human Chemokine Receptor CXCR4.Molecular pharmacology · 2020Article
- TRUPATH, an open-source biosensor platform for interrogating the GPCR transducerome.Nature chemical biology · 2020Article
- Not So Dry After All: DRY Mutants of the AT1ACS omega · 2020Article
Corrections and comments
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Authors and funding
7 authors at 4 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundGq is a heterotrimeric G protein that plays an important role in numerous physiological processes. To delineate the molecular mechanisms and kinetics of signalling through this protein, its activation should be measurable in single living cells. Recently, fluorescence resonance energy transfer (FRET) sensors have been developed for this purpose.
resultsIn this paper, we describe the development of an improved FRET-based Gq activity sensor that consists of a yellow fluorescent protein (YFP)-tagged Gγ2 subunit and a Gαq subunit with an inserted monomeric Turquoise (mTurquoise), the best cyan fluorescent protein variant currently available. This sensor enabled us to determine, for the first time, the kon (2/s) of Gq activation. In addition, we found that the guanine nucleotide exchange factor p63RhoGEF has a profound effect on the number of Gq proteins that become active upon stimulation of endogenous histamine H1 receptors. The sensor was also used to measure ligand-independent activation of the histamine H1 receptor (H1R) upon addition of a hypotonic stimulus.
conclusionsOur observations reveal that the application of a truncated mTurquoise as donor and a YFP-tagged Gγ2 as acceptor in FRET-based Gq activity sensors substantially improves their dynamic range. This optimization enables the real-time single cell quantification of Gq signalling dynamics, the influence of accessory proteins and allows future drug screening applications by virtue of its sensitivity.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.