The DOI matches its Crossref record; 25 of the 25 references it cites were found in a registry or an index.
Novel Single Chain cAMP Sensors for Receptor-induced Signal Propagation
Open accessAbstract
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 beta-adrenergic receptor-induced cAMP signals were observed to travel with high speed ( approximately 40 microm/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.
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Cites
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Metadata
- Authors
- Nikolaev, V. O., Bünemann, M., Hein, L., Hannawacker, A., & Lohse, M. J.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-09
- Volume
- 279
- Issue
- 36
- Pages
- 37215-37218
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1971748110
- Cited by
- 688
- References
- 25
- FWCI
- 14.32
- Type
- journal article
- Language
- en
- Open access
- hybrid
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