Journal article
FRET-based cyclic GMP biosensors measure low cGMP concentrations in cardiomyocytes and neurons
- Abstract:
- Several FRET (fluorescence resonance energy transfer)-based biosensors for intracellular detection of cyclic nucleotides have been designed in the past decade. However, few such biosensors are available for cGMP, and even fewer that detect low nanomolar cGMP concentrations. Our aim was to develop a FRET-based cGMP biosensor with high affinity for cGMP as a tool for intracellular signaling studies. We used the carboxyl-terminal cyclic nucleotide binding domain of Plasmodium falciparum cGMP-dependent protein kinase (PKG) flanked by different FRET pairs to generate two cGMP biosensors (Yellow PfPKG and Red PfPKG). Here, we report that these cGMP biosensors display high affinity for cGMP (EC50 of 23 ± 3 nM) and detect cGMP produced through soluble guanylyl cyclase and guanylyl cyclase A in stellate ganglion neurons and guanylyl cyclase B in cardiomyocytes. These biosensors are therefore optimal tools for real-time measurements of low concentrations of cGMP in living cells.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, 1.5MB, Terms of use)
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- Publisher copy:
- 10.1038/s42003-019-0641-x
Authors
- Publisher:
- Springer Nature
- Journal:
- Communications Biology More from this journal
- Volume:
- 2
- Article number:
- 394
- Publication date:
- 2019-10-29
- Acceptance date:
- 2019-10-02
- DOI:
- EISSN:
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2399-3642
- ISSN:
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2399-3642
- Pmid:
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31701023
- Language:
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English
- Keywords:
- Pubs id:
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1070562
- Local pid:
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pubs:1070562
- Deposit date:
-
2020-06-02
Terms of use
- Copyright holder:
- Calamera et al.
- Copyright date:
- 2019
- Rights statement:
- © The Author(s) 2019. Open Access: This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
- Licence:
- CC Attribution (CC BY)
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