ArticleFASEB journal : official publication of the Federation of American Societies for Experimental Biology2014
Single-fluorophore biosensors based on conformation-sensitive GFP variants.
Article in FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed, 24 citations in OpenAlex.
- A Constellation of Fluorescent Biosensors to Illuminate the Galaxy of Protein Kinases.Chembiochem : a European journal of chemical biology · 2026Review
- Structural characterization of green fluorescent protein in the I-state.Scientific reports · 2024Article
- GRID1/GluD1 homozygous variants linked to intellectual disability and spastic paraplegia impair mGlu1/5 receptor signaling and excitatory synapses.Molecular psychiatry · 2024Article
- GEM-IL: A highly responsive fluorescent lactate indicator.Cell reports methods · 2021Article
- Genetically encoded sensors towards imaging cAMP and PKA activity in vivo.Journal of neuroscience methods · 2021Review
- Review
- Combined Optogenetic Approaches Reveal Quantitative Dynamics of Endogenous Noradrenergic Transmission in the Brain.iScience · 2020Article
- Genetically Encoded Fluorescent Biosensors Illuminate the Spatiotemporal Regulation of Signaling Networks.Chemical reviews · 2018Review
- GluD1, linked to schizophrenia, controls the burst firing of dopamine neurons.Molecular psychiatry · 2018Article
- Cardiac optogenetics: using light to monitor cardiac physiology.Basic research in cardiology · 2017Review
- Active Dentate Granule Cells Encode Experience to Promote the Addition of Adult-Born Hippocampal Neurons.The Journal of neuroscience : the official journal of the Society for Neuroscience · 2017Article
- Cytosolic NADH-NAD(+) Redox Visualized in Brain Slices by Two-Photon Fluorescence Lifetime Biosensor Imaging.Antioxidants & redox signaling · 2016Article
- Fluorescent proteins as genetically encoded FRET biosensors in life sciences.Sensors (Basel, Switzerland) · 2015Review
- Genetically-encoded tools for cAMP probing and modulation in living systems.Frontiers in pharmacology · 2015Review
- A PKA activity sensor for quantitative analysis of endogenous GPCR signaling via 2-photon FRET-FLIM imaging.Frontiers in pharmacology · 2014Article
- Noradrenalin and dopamine receptors both control cAMP-PKA signaling throughout the cerebral cortex.Frontiers in cellular neuroscience · 2014Article
- Visualization of cyclic nucleotide dynamics in neurons.Frontiers in cellular neuroscience · 2014Review
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The β-strands of GFP form a rigid barrel that protects the chromophore from external influence. Herein, we identified specific mutations in β-strand 7 that render the chromophore sensitive to interactions of GFP with another protein domain. In the process of converting the FRET-based protein kinase A (PKA) sensor AKAR2 into a single-wavelength PKA sensor containing a GFP and a quencher, we discovered that the quencher was not required and that the sensor response relied on changes in GFP intrinsic fluorescence. The identified mutations in β-strand 7 render GFP fluorescence intensity and lifetime sensitive to conformational changes of the PKA-sensing domain. In addition, sensors engineered from the GCaMP2 calcium indicator to incorporate a conformation-sensitive GFP (csGFP) exhibited calcium-dependent fluorescence changes. We further demonstrate that single GFP sensors report PKA dynamics in dendritic spines of neurons from brain slices on 2-photon imaging with a high signal-to-baseline ratio and minimal photobleaching. The susceptibility of GFP variants to dynamic interactions with other protein domains provides a new approach to generate single wavelength biosensors for high-resolution imaging.
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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.