ArticlePLoS computational biology2008
The spatiotemporal pattern of Src activation at lipid rafts revealed by diffusion-corrected FRET imaging.
Article in PLoS computational biology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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46 citing papers in PubMed, 63 citations in OpenAlex.
- Beyond analytic solution: Analysis of FRAP experiments by spatial simulation of the forward problem.Biophysical journal · 2023Article
- Fluorescence resonance energy transfer (FRET) spatiotemporal mapping of atypical P38 reveals an endosomal and cytosolic spatial bias.Scientific reports · 2023Article
- Luminescence lifetime imaging of three-dimensional biological objects.Journal of cell science · 2021Article
- Mechanobiology of dynamic enzyme systems.APL bioengineering · 2020Article
- Article
- Biophysical basis underlying dynamic Lck activation visualized by ZapLck FRET biosensor.Science advances · 2019Article
- Sensitive FRET Biosensor Reveals Fyn Kinase Regulation by Submembrane Localization.ACS sensors · 2019Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- Article
- Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface.Cell chemical biology · 2018Article
- Epidermal growth factor receptor and integrins control force-dependent vinculin recruitment to E-cadherin junctions.Journal of cell science · 2018Article
- βBiophysical journal · 2017Article
- Effect of probe diffusion on the SOFI imaging accuracy.Scientific reports · 2017Article
- CaFrontiers in oncology · 2017Review
- Imaging Spatiotemporal Activities of ZAP-70 in Live T Cells Using a FRET-Based Biosensor.Annals of biomedical engineering · 2016Article
- In-situ coupling between kinase activities and protein dynamics within single focal adhesions.Scientific reports · 2016Article
- Toluene disruption of the functions of L1 cell adhesion molecule at concentrations associated with occupational exposures.Pediatric research · 2016Article
- Novel Bioluminescent Activatable Reporter for Src Tyrosine Kinase Activity in Living Mice.Theranostics · 2016Article
- Activatable and Cell-Penetrable Multiplex FRET Nanosensor for Profiling MT1-MMP Activity in Single Cancer Cells.Nano letters · 2015Article
- Subcellular and Dynamic Coordination between Src Activity and Cell Protrusion in Microenvironment.Scientific reports · 2015Article
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
Abstract
Genetically encoded biosensors based on fluorescence resonance energy transfer (FRET) have been widely applied to visualize the molecular activity in live cells with high spatiotemporal resolution. However, the rapid diffusion of biosensor proteins hinders a precise reconstruction of the actual molecular activation map. Based on fluorescence recovery after photobleaching (FRAP) experiments, we have developed a finite element (FE) method to analyze, simulate, and subtract the diffusion effect of mobile biosensors. This method has been applied to analyze the mobility of Src FRET biosensors engineered to reside at different subcompartments in live cells. The results indicate that the Src biosensor located in the cytoplasm moves 4-8 folds faster (0.93+/-0.06 microm(2)/sec) than those anchored on different compartments in plasma membrane (at lipid raft: 0.11+/-0.01 microm(2)/sec and outside: 0.18+/-0.02 microm(2)/sec). The mobility of biosensor at lipid rafts is slower than that outside of lipid rafts and is dominated by two-dimensional diffusion. When this diffusion effect was subtracted from the FRET ratio images, high Src activity at lipid rafts was observed at clustered regions proximal to the cell periphery, which remained relatively stationary upon epidermal growth factor (EGF) stimulation. This result suggests that EGF induced a Src activation at lipid rafts with well-coordinated spatiotemporal patterns. Our FE-based method also provides an integrated platform of image analysis for studying molecular mobility and reconstructing the spatiotemporal activation maps of signaling molecules in live cells.
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