Novel Single Chain cAMP Sensors for Receptor-induced Signal Propagation*♦

By V. Nikolaev, M. Bünemann, L. Hein, Annette Hannawacker, M. Lohse. Journal of Biological Chemistry, 2004

701 citations recorded by the source corpus.

cAMP is a universal second messenger of many G-protein-coupled receptors and regulates a wide variety of cellular events. cAMP exerts its effects via cAMP-dependent protein kinase (PKA), cAMP-gated ion channels, and two isoforms of exchange protein directly activated by cAMP (Epac). Here we report the development of novel fluorescent indicators for cAMP based on the cAMP-binding domains of Epac and PKA. Fluorescence resonance energy transfer between variants of green fluorescent protein (enhanced cyan fluorescent protein and enhanced yellow fluorescent protein) fused directly to the cAMP-binding domains was used to analyze spatial and temporal aspects of cAMP-signaling in different cells. In contrast to previously developed PKA-based indicators, these probes are comprised of only a single binding site lacking cooperativity, catalytic properties, and interactions with other proteins and thereby allow us to easily image free intracellular cAMP and rapid signaling events. Rapid β-adrenergic receptor-induced cAMP signals were observed to travel with high speed (≈40 μm/s) throughout the entire cell body of hippocampal neurons and peritoneal macrophages. The developed indicators could be ubiquitously applied to studying cAMP, its physiological role and spatio-temporal regulation.

Subfields: Cellular and Molecular Neuroscience, Molecular Biology

View Paper: https://www.semanticscholar.org/paper/9d3e46badd8a9a137d7254fd240a36b926596d99

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Cellular and Molecular Neuroscience

In Molecular Biology

Citations in the corpus

Dynamics of learning-related cAMP signaling and stimulus integration in the Drosophila olfactory pathway. (2009, positive): “, 1996, 1998), we turned to cAMP imaging using the genetically encoded optical reporter epac1-camps (Nikolaev et al., 2004; Shafer et al., 2008).”
A single pair of neurons links sleep to memory consolidation in Drosophila melanogaster (2015, methodology): “, 2011) to drive expression of Epac1-camps (EPAC) (Nikolaev et al., 2004; Shafer et al., 2008), a FRET-based cyclic nucleotide sensor.”
Analysis of functional neuronal connectivity in the Drosophila brain. (2012, methodology): “Epac1-camps (Nikolaev et al. 2004) was sequentially digested from the pUAST-Epac1-camps plasmid (Shafer et al. 2008) using XhoI and BglII, and PCR purified.”