ArticleACS sensors2019
Sensitive FRET Biosensor Reveals Fyn Kinase Regulation by Submembrane Localization.
Article in ACS sensors, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 30 citations in OpenAlex.
- Molecular Spies in Action: Genetically Encoded Fluorescent Biosensors Light up Cellular Signals.Chemical reviews · 2024Review
- Light-gated integrator for highlighting kinase activity in living cells.Nature communications · 2024Article
- Article
- Characterization of PDGF-Induced Subcellular Calcium Regulation through Calcium Channels in Airway Smooth Muscle Cells by FRET Biosensors.Biosensors · 2024Article
- Light-gated Integrator for Highlighting Kinase Activity in Living Cells.bioRxiv : the preprint server for biology · 2024Article
- FRET Visualization of Cyclic Stretch-Activated ERKFrontiers in cell and developmental biology · 2022Article
- Integration of FRET and sequencing to engineer kinase biosensors from mammalian cell libraries.Nature communications · 2021Article
- Bioengineering tools for probing intracellular events in T lymphocytes.WIREs mechanisms of disease · 2021Review
- A Perspective of Epigenetic Regulation in Radiotherapy.Frontiers in cell and developmental biology · 2021Review
- Cellular Plasticity during Metastasis: New Insights Provided by Intravital Microscopy.Cold Spring Harbor perspectives in medicine · 2020Review
- Optogenetic Control for Investigating Subcellular Localization of Fyn Kinase Activity in Single Live Cells.Journal of molecular biology · 2020Article
- Phosphorylation-Dependent SERS Readout for Activity Assay of Protein Kinase A in Cell Extracts.Nanomaterials (Basel, Switzerland) · 2020Article
- Article
- Live-cell monitoring of protein localization to membrane rafts using protein-fragment complementation.Bioscience reports · 2020Article
- Article
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Authors and funding
12 authors at 2 institutions in 1 country.
Funding
Abstract
Fyn kinase plays crucial roles in hematology and T cell signaling; however, there are currently limited tools to visualize the dynamic Fyn activity in live cells. Here we developed and characterized a highly sensitive Fyn biosensor based on fluorescence resonance energy transfer (FRET) to monitor Fyn kinase activity in live cells. Our results show that Fyn kinase activity can be induced in both mouse embryonic fibroblasts (MEFs) and T cells by ligand engagement. Two different motifs were further introduced to target the biosensor at the cellular membrane microdomains in MEFs, revealing that the Fyn-tagged biosensor had 70% greater response to growth factor stimulation than the Lyn-tagged version. This suggests that the plasma membrane microdomains can be categorized into different functional subdomains. Further experiments show that while the membrane accessibility is necessary for Fyn activation, the localization of Fyn outside of its microdomains causes its hyperactivity, indicating that membrane microdomains provide a suppressive microenvironment for Fyn regulation in MEFs. Interestingly, a relatively high Fyn activity can be observed at perinuclear regions, further supporting the notion that the membrane microenvironment has a significant impact on the local molecular functions. Our work hence highlights a novel Fyn FRET biosensor for live cell imaging and its application in revealing an intricate submembrane regulation of Fyn in live MEFs.
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