Titlebook: cAMP Signaling; Methods and Protocol Manuela Zaccolo Book 2015 Springer Science+Business Media New York 2015 Adenosine 3′,5′-monophosphate.

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https://doi.org/10.1007/978-1-4939-2537-7Adenosine 3′,5′-monophosphate; Cellular function; Disease mechanisms; Protein kinase A (PKA); Signaling

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978-1-4939-4950-2Springer Science+Business Media New York 2015

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https://doi.org/10.1007/978-3-319-63321-3A), can be measured in living cells, thanks to genetically encoded probes based on fluorescence resonance energy transfer (FRET). While these biosensors enable the assessment of cAMP or PKA activity with great spatial and temporal resolution, concomitant events triggered by the same stimuli at the s

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Bernardo Cardoso,Jorge Ferraz de Abreuwnstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (

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Applications and Usability of Interactive TVcAMP) signaling and compartmentalization in living cells. These sensors allow estimation of relative changes of cAMP levels in real-time and intact cells. However, one of their major shortcomings is that they do not easily allow direct measurement of cAMP concentrations. This is mainly due to the fa

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Applications and Usability of Interactive TV of local concentrations of second messengers in living cells. Nowadays, the availability of a number of 3D structures of cyclic nucleotide-binding domains (CNBD) undergoing conformational transitions upon cAMP binding, along with computational tools, can be exploited for the design of novel or impr

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Applications and Usability of Interactive TVch to measure localized cAMP signals. However, given the low signal-to-noise ratio of most FRET probes and the dynamic nature of the intracellular environment, there have been marked limitations in the ability to use FRET probes to study localized signaling events within the same cell. Here, we outl

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