FRET-based cyclic GMP biosensors measure low cGMP concentrations in cardiomyocytes and neurons.

Autor: Calamera G; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway., Li D; 3Department of Physiology, Anatomy and Genetics, Oxford University, Oxford, UK., Ulsund AH; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway., Kim JJ; 4Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX USA., Neely OC; 3Department of Physiology, Anatomy and Genetics, Oxford University, Oxford, UK., Moltzau LR; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway., Bjørnerem M; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway., Paterson D; 3Department of Physiology, Anatomy and Genetics, Oxford University, Oxford, UK., Kim C; 4Department of Pharmacology and Chemical Biology, Baylor College of Medicine, Houston, TX USA.; 5Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX USA., Levy FO; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway., Andressen KW; 1Department of Pharmacology, Institute of Clinical Medicine, University of Oslo and Oslo University Hospital, Oslo, Norway.; 2Center for Heart Failure Research, University of Oslo and Oslo University Hospital, Oslo, Norway.
Jazyk: angličtina
Zdroj: Communications biology [Commun Biol] 2019 Oct 29; Vol. 2, pp. 394. Date of Electronic Publication: 2019 Oct 29 (Print Publication: 2019).
DOI: 10.1038/s42003-019-0641-x
Abstrakt: 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 Pf PKG and Red Pf PKG). Here, we report that these cGMP biosensors display high affinity for cGMP (EC 50 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.
Competing Interests: Competing interestsThe authors have no competing interests to declare.
(© The Author(s) 2019.)
Databáze: MEDLINE
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