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
- ER McGlone, et al. Hepatocyte cholesterol content modulates glucagon receptor signaling. Molecular Metabolism. September 2022.
- B. Barsi-Rhyne, et al. Discrete GPCR-triggered endocytic modes enable β-arrestins to flexible regulate cell signaling. bioRxiv. July 2022.
- J. Xu, J. Pluznick. Key amino acids alter activity and trafficking of a well-conserved olfactory receptor. Cell Physiology. June 2022.
- C. Zhang, et al. A brainstem circuit for nausea suppression. Cell Reports. June 2022.
- J. Hansen, et al. A cAMP signalome in primary cilia drives gene expression and kidney cyst formation. EMBO Reports. June 2022.
- S. Ansari, et al. Morphogen Directed Coordination of GPCR Activity Promotes Primary Cilium Function for Downstream Signaling. bioRxiv. May 2022.
- E. Porpiglia, et al. Elevated CD47 is a hallmark of dysfunctional aged muscle stem cells that can be targeted to augment regeneration. bioRxiv. April 2022.
- S. Bitsi, et al. Divergent acute versus prolonged in vivo GLP-1R responses in β-arrestin 2-deleted primary beta cells. bioRxiv. April 2022.
- Y. Mizobuchi, et al. Ketamine Improves Desensitization of μ-Opioid Receptors Induced by Repeated Treatment with Fentanyl but Not with Morphine. biomolecules. March 2022.
- B. Polacco, et al. Profiling the diversity of agonist-selective effects on the proximal proteome environment of G protein-coupled receptors. bioRxiv. March 2022.
- D. Lovinger, et al. Local modulation by presynaptic receptors controls neuronal communication and behavior. Nature Reviews Neuroscience. February 2022.
- F. De Logu, et al. Schwann cell endosome CGRP signals elicit peri orbital mechanical allodynia in mice. Nature Communications. February 2022.
- A. Lutas, et al. History-dependent dopamine release increases cAMP levels in most basal amygdala glutamatergic neurons to control learning. Cell Reports. January 2022.
- G. Sancar, et al. FGF1 and insulin control lipolysis by convergent pathways. Cell Metabolism. January 2022.
- S. Hoare, et al. Quantifying the Kinetics of Signaling and Arrestin Recruitment by Nervous System G-Protein Coupled Receptors. Frontiers in Cellular Neuroscience. January 2022.
- J. H. Cho, et al. Islet primary cilia motility controls insulin secretion. bioRxiv. December 2021.
- Y. Karasawa, et al. In Vitro Analyses of Spinach-Derived Opioid Peptides, Rubiscolins: Receptor Selectivity and Intracellular Activities through G-protein- and β-arrestin-Mediated Pathways. Molecules. October 2021.
- A. White, et al. Spatial Bias in cAMP generation determines biological responses to PTH type 1 receptor activation. Science Signaling. October 2021.
- J. Janetzko, et al. Membrane phosphoinositides stabilize GPCR-arrestin complexes and offer temporal control of complex assembly and dynamics. bioRxiv. October 2021.
- S. Hoare, T. Hughes. Biosensor Assays for Measuring the Kinetics of G-Protein and Arrestin-Mediated Signaling in Live Cells. The Assay Guidance Manual. September 2021.
- R. Carey, et al. HSP90 Function is Required for T2R Bitter Taste Receptor Nitric Oxide Production and Innate Immune Responses in Human Airway Epithelial Cells and Macrophages. Cells. April 2022. (bioRxiv)
- D. McMahon, et al. The bitter end: T2R bitter receptor agonists elevate nuclear calcium and induce apoptosis in non-ciliated airway epithelial cells. Cell Calcium. January 2022. (bioRxiv)
- 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. Nuriya, et al. Alkyne-Tagged Dopamines as Versatile Analogue Probes for Dopaminergic System Analysis. Analytical Chemistry. July 2021.
- M. Thomas, et al. Optically activated, customizable, excitable cells. PLOS One. December 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 Frontiers Cellular Neuroscience. December 2019.
- N. Naim et al. Luminescence-activated nucleotide cyclase regulates spatial and temporal cAMP synthesis. Journal of Biological Chemistry. December 2018.
- Y.A.B. Sierra, et al. Potassium channel-based optogenetic silencing. Nature Communications. November 2018.
