Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of Phosphodiesterase-Protein Kinase Complexes as Novel Targets for Discovery of Inhibitors with Enhanced Specificity

Version 1 : Received: 5 April 2021 / Approved: 6 April 2021 / Online: 6 April 2021 (12:07:58 CEST)

How to cite: Tulsian, N.K.; Sin, V.J.; Anand, G.S.; Koh, H. Development of Phosphodiesterase-Protein Kinase Complexes as Novel Targets for Discovery of Inhibitors with Enhanced Specificity. Preprints 2021, 2021040174 (doi: 10.20944/preprints202104.0174.v1). Tulsian, N.K.; Sin, V.J.; Anand, G.S.; Koh, H. Development of Phosphodiesterase-Protein Kinase Complexes as Novel Targets for Discovery of Inhibitors with Enhanced Specificity. Preprints 2021, 2021040174 (doi: 10.20944/preprints202104.0174.v1).

Abstract

Phosphodiesterases (PDEs) hydrolyze cyclic nucleotides to modulate multiple signaling events in cells. PDEs are recognized to actively associate with cyclic nucleotide receptors (Protein Kinases, PK) in larger macromolecular assemblies referred to as signalosomes. Complexation of PDEs with PK generates an expanded active site which enhances PDE activity. This facilitates signalosome-associated PDEs to preferentially catalyze active hydrolysis of cyclic nucleotides bound to PK, and aid in signal termination. PDEs are important drug targets and current strategies for inhibitor discovery are based entirely on targeting conserved PDE catalytic domains. This often results in inhibitors with cross-reactivity amongst closely related PDEs and attendant unwanted side effects. Here, our approach targets PDE-PK complexes as they would occur in signalosomes, thereby offering greater specificity. Our developed fluorescence polarization assay has been adapted to identify inhibitors that block cyclic nucleotide pockets in PDE-PK complexes in one mode, and disrupt protein-protein interactions between PDEs and cyclic nucleotide activating protein kinases in a second mode. We tested this approach with three different systems: cAMP-specific PDE8-PKAR, cGMP-specific PDE5-PKG and dual-specificity RegA-RD complexes and ranked inhibitors according to their inhibition potency. Targeting PDE-PK complexes offers biochemical tools for describing the exquisite specificity of cyclic nucleotide signaling networks in cells.

Subject Areas

Phosphodiesterase (PDE); natural products; inhibitors; Protein Kinase; selectivity; fluorescence polarization

Comments (0)

We encourage comments and feedback from a broad range of readers. See criteria for comments and our diversity statement.

Leave a public comment
Send a private comment to the author(s)
Views 0
Downloads 0
Comments 0
Metrics 0


×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here.