PDE Biology
Cyclic nucleotide phosphodiesterases (PDEs) are important drug targets with unique tissue distribution and functional properties. This family of enzymes is key in the regulation of cyclic nucleotide second messenger levels because they degrade both cAMP and cGMP. Some PDEs hydrolyze both cAMP and cGMP, while others are either cAMP or cGMP hydrolases. Selective PDE inhibitors block the activity of specific PDEs and are used to treat a variety of diseases. Non-selective PDE inhibitors include IBMX and caffeine, both of which are important consumables in basic research laboratories.
Detecting cAMP
cAMP response to isoproterenol followed by inhibition of cAMP production with quinpirole. Detected with the cADDis cAMP assay.
PDE Selectivity Assay
An Optogenetic Tool for Monitoring PDE Rates
The blue-light activated adenylyl cyclase bPAC (#V0100N) can be co-expressed with R-cADDis (#U0200R), a red fluorescent biosensor for cAMP. A two second pulse of blue light activates bPAC to stimulate cAMP production. cAMP degradation by PDE is monitored in real time using R-cADDis. Read about how this method can be used to study the role of PDE activity in neurodegeneration: Live-Cell Assays for Cell Stress Responses Reveal New Patterns of Cell Signaling Caused by Mutations in Rhodopsin, α-Synuclein and TDP-43.
Simple Protocol
Simple, step-by-step protocols for optimizing cAMP and cGMP assays are available online. These assays are non-FRET and cell lysis, enzymes, and co-factors are not necessary. Add our sensors to your cells, incubate, add drug, and measure fluorescence changes.
Sound Intriguing? Get more details on our PDE Biology Assays:
cADDis cAMP Assay
GENIe cGMP Assay
bPAC in BacMam
Recent Publications
- I. Gapallawa, et al. Bitter taste receptors stimulate phagocytosis in human macrophages through calcium, nitric oxide, and cyclic-GMP signaling Cell. Mol. Life Sci. Feb, 2020 (bioRxiv)
- D. McMahon, et al. Neuropeptide regulation of secretion and inflammation in human airway gland serous cells (bioRxiv)
- R. Forteza, et al. Glucocorticoids and myosin5b loss of function induce heightened PKA signaling in addition to membrane traffic defects Molecular Biology of the Cell. Dec. 2019
- Nunez, F.J., Johnstone, T.B. et al. Glucocorticoids rapidly activate cAMP production via Gαs to initiate non‐genomic signaling that contributes to one‐third of their canonical genomic effects FASEB Journal, Dec. 2019
- Nunez, F.J., Schulte, N.A., Fogel, D.M., et al. Agonist-specific desensitization of PGE2-stimulated cAMP signaling due to upregulated phosphodiesterase expression in human lung fibroblasts Naunyn-Schmiedeberg’s Arch Pharmacol Dec. 2019
- K. Harlen, et al. Live-Cell Assays for Cell Stress Responses Reveal New Patterns of Cell Signaling Caused by Mutations in Rhodopsin, α-Synuclein and TDP-43 Front. Cell. Neurosci.,December 2019
- K. Hilgendorf, et al. Omega-3 Fatty Acids Activate Ciliary FFAR4 to Control Adipogenesis Cell November, 2019
- R. Forteza, et al. Glucocorticoids and Myosin5b loss-of-function induce heightened PKA signaling in addition to membrane traffic defects MBoC Oct, 2019
- T. Baldwin, et al. Insights into the Regulatory Properties of Human Adenylyl Cyclase Type 9 Molecular Pharmacology. April, 2019
- T. Togo Autocrine purinergic signaling stimulated by cell membrane disruption is involved in both cell membrane repair and adaptive response in MDCK cells Biochem & Biophysical Research Comm. March, 2019
- C. Ojiaku, et al. Transforming Growth Factor-β1 Decreases β2-Agonist–induced Relaxation in Human Airway Smooth Muscle ATSJournas, Dec 2018
- Y.A.B. Sierra, et al. Potassium channel-based optogenetic silencing. Nature Communications Nov. 5 2018.
- X. Chen, et al. Phenylephrine, a common cold remedy active ingredient, suppresses uterine contractions through cAMP signalling. Scientific Reports Aug. 2018.
- P. Tewson, et al. Assay for Detecting Gαi-Mediated Decreases in cAMP in Living Cells. SLAS. July 10, 2018. Request Full Text PDF
- T. Buranda, et al. A High-Throughput Flow Cytometry Screen Identifies Molecules that Inhibit Hantavirus Cell Entry. SLAS Discovery. April 2, 2018.
- N. H. Wray, et al. NMDAR-independent, cAMP-dependent antidepressant actions of ketamine. Molecular Psychiatry. April 2, 2018.
- H. Zou, et al. PDE8: A Novel Target in Airway Smooth Muscle. American Journal of Respiratory Cell and Molecular Biology. Vol. 58, No. 4. April 01 2018.
- M. Thomas, et al. Toward a High Throughput Screen for Voltage Sensors. 2018.
- T.B. Johnstone, et al. PDE8 is Expressed in Human Airway Smooth Muscle and Selectively Regulates cAMP Signaling by β2AR-AC6. American Journal of Respiratory Cell and Molecular Biology. Dec. 2017.
- K.A. McCrink, et al. β-Arrestin2 Improves Post-Myocardial Infarction Heart Failure via Sarco(endo)plasmic Reticulum Ca2+-ATPase-Dependent Positive Inotropy in Cardiomyocytes. Hypertension. Nov. 2017.
- J. Almaça, et al. Human beta cells produce and release serotonin to inhibit glucagon secretion from alpha cells.Cell Reports, Volume 17, Issue 12, 2016.
- P. Tewson, et al. New DAG and cAMP Sensors Optimized for Live-Cell Assays in Automated Laboratories. J Biomol Screen Dec. 11, 2015. Request Full Text PDF
- B.S. Moore, et al. Cilia have high cAMP levels that are inhibited by Sonic Hedgehog-regulated calcium dynamics. PNAS Nov. 15, 2016.
- I. Gapallawa, et al. Bitter taste receptors stimulate phagocytosis in human macrophages through calcium, nitric oxide, and cyclic-GMP signaling Cell. Mol. Life Sci. Feb, 2020 (bioRxiv)
- M. Thomas, et al. Design for Fast Optogenetic Screen in Mammalian Cells For Next Gen Ca2+ Sensors. BioRxiv. Feb 2020.
- K. Harlen, et al. Live-Cell Assays for Cell Stress Responses Reveal New Patterns of Cell Signaling Caused by Mutations in Rhodopsin, α-Synuclein and TDP-43 Front. Cell. Neurosci.,December 2019.
