ArticleCancer research2010
Simultaneous visualization of protumorigenic Src and MT1-MMP activities with fluorescence resonance energy transfer.
Article in Cancer research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 70 papers.
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Who cites it
70 citing papers in PubMed, 105 citations in OpenAlex.
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- Integration of FRET and sequencing to engineer kinase biosensors from mammalian cell libraries.Nature communications · 2021Article
- Strategies for Multiplexed Biosensor Imaging to Study Intracellular Signaling Networks.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Cellular Plasticity during Metastasis: New Insights Provided by Intravital Microscopy.Cold Spring Harbor perspectives in medicine · 2020Review
- A Novel NIR-FRET Biosensor for Reporting PS/γ-Secretase Activity in Live Cells.Sensors (Basel, Switzerland) · 2020Article
- FRET-Based CaSensors (Basel, Switzerland) · 2020Article
- Application of FRET Biosensors in Mechanobiology and Mechanopharmacological Screening.Frontiers in bioengineering and biotechnology · 2020Review
- Label-Free Assay of Protein Tyrosine Phosphatase Activity in Single Cells.Analytical chemistry · 2019Article
- Discretionary Transduction of MMP-Sensitized Tousled in Head and Neck Cancer.Molecular therapy oncolytics · 2019Article
- Characterization and regulation of MT1-MMP cell surface-associated activity.Chemical biology & drug design · 2019Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- Synergistic Ensemble of Optogenetic Actuators and Dynamic Indicators in Cell Biology.Molecules and cells · 2018Review
- Direct multiplex imaging and optogenetics of Rho GTPases enabled by near-infrared FRET.Nature chemical biology · 2018Article
- Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface.Cell chemical biology · 2018Article
- Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2.Scientific reports · 2017Article
10 more citing papers are in PubMed but not listed here.
Corrections and comments
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Authors and funding
8 authors at 3 institutions in 1 country.
Funding
Abstract
Both Src kinase and membrane type 1 matrix metalloproteinase (MT1-MMP) play critical roles in cancer invasion and metastasis. It is not clear, however, how the spatiotemporal activation of these two critical enzymes is coordinated in response to an oncogenic epithelial growth factor (EGF) stimulation. Here, we have visualized the activities of Src and MT1-MMP concurrently in a single live cell by combining two fluorescence resonance energy transfer (FRET) pairs with distinct spectra: (a) cyan fluorescent protein (CFP) and yellow FP (YFP), and (b) orange FP (mOrange2) and red FP (mCherry). The new FRET pair, mOrange2 and mCherry, was first characterized in vitro and in cultured mammalian cells. When integrated with the CFP/YFP pair, this new pair allowed the revelation of an immediate, rapid, and relatively dispersed Src activity. In contrast, the MT1-MMP activity displayed a slow increase at the cell periphery, although Src was shown to play a role upstream to MT1-MMP globally. This difference in the activation patterns of MT1-MMP and Src in response to EGF is further confirmed using an optimized MT1-MMP biosensor capable of being rapidly cleaved by MT1-MMP. The results indicate that although Src and MT1-MMP act globally in the same signaling pathway, their activations differ in space and time upon EGF stimulation, possibly mediated by different sets of intermediates at different subcellular locations. Our results also showed the potential of mOrange2/mCherry as a new FRET pair, together with the popular variants of CFP and YFP, for the simultaneous visualization of multiple molecular activities in a single live cell.
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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.