ArticleBiophysical journal2008
Quantitative measurement of cAMP concentration using an exchange protein directly activated by a cAMP-based FRET-sensor.
Article in Biophysical journal, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed, 32 citations in OpenAlex.
- Extracellular bimolecular fluorescence complementation for investigating membrane protein dimerization: a proof of concept using class B GPCRs.Bioscience reports · 2024Article
- Computational investigation of the dynamic control of cAMP signaling by PDE4 isoform types.Biophysical journal · 2022Article
- Green fluorescent cAMP indicator of high speed and specificity suitable for neuronal live-cell imaging.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Absolute measurement of cellular activities using photochromic single-fluorophore biosensors and intermittent quantification.Nature communications · 2022Article
- Distinct temporal integration of noradrenaline signaling by astrocytic second messengers during vigilance.Nature communications · 2020Article
- Intracellular cAMP Sensor EPAC: Physiology, Pathophysiology, and Therapeutics Development.Physiological reviews · 2018Review
- Energy transfer from an individual silica nanoparticle to graphene quantum dots and resulting enhancement of photodetector responsivity.Scientific reports · 2016Article
- Two New FRET Imaging Measures: Linearly Proportional to and Highly Contrasting the Fraction of Active Molecules.PloS one · 2016Article
- From intracellular signaling to population oscillations: bridging size- and time-scales in collective behavior.Molecular systems biology · 2015Article
- Activation of soluble adenylyl cyclase protects against secretagogue stimulated zymogen activation in rat pancreaic acinar cells.PloS one · 2012Article
- An ion-insensitive cAMP biosensor for long term quantitative ratiometric fluorescence resonance energy transfer (FRET) measurements under variable physiological conditions.The Journal of biological chemistry · 2011Article
- Signal/noise analysis of FRET-based sensors.Biophysical journal · 2010Article
- Long wavelength fluorescence lifetime standards for front-face fluorometry.Journal of fluorescence · 2010Article
- H(2)O(2)-mediated modulation of cytosolic signaling and organelle function in rat hippocampus.Pflugers Archiv : European journal of physiology · 2009Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Förster resonance energy transfer (FRET)-based biosensors for the quantitative analysis of intracellular signaling, including sensors for monitoring cyclic adenosine monophosphate (cAMP), are of increasing interest. The measurement of the donor/acceptor emission ratio in tandem biosensors excited at the donor excitation wavelength is a commonly used technique. A general problem, however, is that this ratio varies not only with the changes in cAMP concentration but also with the changes of the ionic environment or other factors affecting the folding probability of the fluorophores. Here, we use a spectral FRET analysis on the basis of two excitation wavelengths to obtain a reliable measure of the absolute cAMP concentrations with high temporal and spatial resolution by using an "exchange protein directly activated by cAMP". In this approach, FRET analysis is simplified and does not require additional calibration routines. The change in FRET efficiency (E) of the biosensor caused by [cAMP] changes was determined as DeltaE = 15%, whereas E varies between 35% at low and 20% at high [cAMP], allowing quantitative measurement of cAMP concentration in the range from 150 nM to 15 microM. The method described is also suitable for other FRET-based biosensors with a 1:1 donor/acceptor stoichiometry. As a proof of principle, we measured the specially resolved cAMP concentration within living cells and determined the dynamic changes of cAMP levels after stimulation of the Gs-coupled serotonin receptor subtype 7 (5-HT7).
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