Identification of PDE4B Over 4D subtype-selective inhibitors revealing an unprecedented binding mode

Kranz, Michael; Wall, Michael; Evans, Brian; Miah, Afjal H.; Ballantine, Stuart P.; Delves, C. J.; Dombroski, Brian; Gross, Janet L.; Schneck, Jessica L.; Villa, James P.; Neu, Margarete; Somers, Don O.

doi:10.1016/j.bmc.2009.03.061

Cited by 53 publications

(40 citation statements)

References 18 publications

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“…Rol, rolipram; Fsk, forskolin. compartment and that the above data are not the result of longterm compensation for the absence of PDE4B, we used an alternative strategy to probe local PDE4B function. PDE4B-selective compounds have been developed to improve the efficacy and safety of PDE4 inhibition (Donnell et al, 2010;Kranz et al, 2009). We therefore used a PDE4B-selective inhibitor (supplementary material Fig.…”

Section: Pde4b Functions In a Confined Compartment Near The Sarcolemmmentioning

confidence: 99%

PDE4B mediates local feedback regulation of β1-adrenergic cAMP signaling in a sarcolemmal compartment of cardiac myocytes

Mika

Richter

Westenbroek

et al. 2014

Journal of Cell Science

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Multiple cAMP phosphodiesterase (PDE) isoforms play divergent roles in cardiac homeostasis but the molecular basis for their nonredundant function remains poorly understood. Here, we report a novel role for the PDE4B isoform in b-adrenergic (bAR) signaling in the heart. Genetic ablation of PDE4B disrupted bAR-induced cAMP transients, as measured by FRETsensors, at the sarcolemma but not in the bulk cytosol of cardiomyocytes. This effect was further restricted to a subsarcolemmal compartment because PDE4B regulates b 1 AR-, but not b 2 AR-or PGE2-induced responses. The spatially restricted function of PDE4B was confirmed by its selective effects on PKA-mediated phosphorylation patterns. PDE4B limited the PKA-mediated phosphorylation of key players in excitationcontraction coupling that reside in the sarcolemmal compartment, including L-type Ca 2+ channels and ryanodine receptors, but not phosphorylation of distal cytosolic proteins. b 1 AR-but not b 2 ARligation induced PKA-dependent activation of PDE4B and interruption of this negative feedback with PKA inhibitors increased sarcolemmal cAMP. Thus, PDE4B mediates a crucial PKAdependent feedback that controls b 1 AR-dependent cAMP signals in a restricted subsarcolemmal domain. Disruption of this feedback augments local cAMP/PKA signals, leading to an increased intracellular Ca 2+ level and contraction rate.

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Section: Pde4b Functions In a Confined Compartment Near The Sarcolemmmentioning

confidence: 99%

PDE4B mediates local feedback regulation of β1-adrenergic cAMP signaling in a sarcolemmal compartment of cardiac myocytes

Mika

Richter

Westenbroek

et al. 2014

Journal of Cell Science

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“…However, there are at least two other possible mechanisms of regulation that apply to short and supershort as well as long isoforms. First, in some crystal structures of the catalytic domain of PDE4B and PDE4D, a short helical segment of the C-terminal region (not visible in the PDE4B cryst crystal structure), has been seen to interact with the catalytic site in a manner similar to the interaction of the regulatory helix seen in this structure [PDB ID codes 1F0J (35), 1XM6 (24), 3HMV (36), and 4MYQ (37)]. The short linker between the C terminus of the catalytic domain and the C-terminal helix requires that this interaction has to be within one subunit, and does not require dimerization.…”

Section: Mapping Pde4d Gene Mutations Found In Acrodysostosis Patientsmentioning

confidence: 99%

Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4

Cedervall

Aulabaugh

Geoghegan

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Phosphodiesterase 4 (PDE4) is an essential contributor to intracellular signaling and an important drug target. The four members of this enzyme family (PDE4A to -D) are functional dimers in which each subunit contains two upstream conserved regions (UCR), UCR1 and -2, which precede the C-terminal catalytic domain. Alternative promoters, transcriptional start sites, and mRNA splicing lead to the existence of over 25 variants of PDE4, broadly classified as long, short, and supershort forms. We report the X-ray crystal structure of long form PDE4B containing UCR1, UCR2, and the catalytic domain, crystallized as a dimer in which a disulfide bond cross-links cysteines engineered into UCR2 and the catalytic domain. Biochemical and mass spectrometric analyses showed that the UCR2-catalytic domain interaction occurs in trans, and established that this interaction regulates the catalytic activity of PDE4. By elucidating the key structural determinants of dimerization, we show that only long forms of PDE4 can be regulated by this mechanism. The results also provide a structural basis for the long-standing observation of high-and low-affinity binding sites for the prototypic inhibitor rolipram.T he diverse, pivotal roles of cAMP and cGMP (1) in mammalian intracellular signaling include acting as "second messengers" of signals delivered through G protein-coupled receptors (2), regulating the opening and closing of ligand-gated ion-channels (3), activating guanine-nucleotide exchange factors (4), and activating kinase signaling cascades (5). Responsibility for degrading these 3′,5′-cyclic nucleotides, and thereby limiting the duration and amplitude of the signals that they convey, falls to enzymes of the intracellular phosphodiesterase family (PDEs). The significance of these enzymes in cellular control makes them important targets of medicinal discovery, and maximizing our understanding of their respective structures and activities is an urgent priority (6).Mammalian PDEs are encoded by 21 genes and divided into 11 families (PDE1 to -11) defined by their amino acid sequences, with supporting distinctions based on pharmacologic and kinetic properties. Among the most intriguing of these families is PDE4, the four members (PDE4A to -D) of which have high selectivity for cAMP over cGMP as substrate and are characteristically sensitive to inhibition by rolipram. PDE4 inhibition has been pursued as a therapeutic target for over 30 y, since the first discovery of the potential efficacy of rolipram in treating clinical depression (7). Two different PDE4 inhibitors, roflumilast (Daxas) and apremilast (Otezla), are Food and Drug Administration-approved therapies for exacerbations of chronic obstructive pulmonary disease and psoriatic arthritis, respectively, and several others are in clinical trials for a variety of indications.Because of alternative promoters/start sites and variable mRNA splicing, transcription from the four PDE4 genes results in the expression of more than 25 different isoforms of PDE4. Each isoform has a unique N...

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“…Other crystal structures of the catalytic domain of PDE4B (1F0J [13], 1XM6 [43], 3HMV [44], 3KKT (unpublished)) have also reported a similar C-terminal helix closing over the active site (Figure 3.6). These structures are the exceptions, as in the majority of PDE4B catalytic domain structures the C-terminal residues are unstructured, and not modeled.…”

Section: Crystal Structure Of Regulatory Domains Of Pde4mentioning

confidence: 89%

PDE4: New Structural Insights into the Regulatory Mechanism and Implications for the Design of Selective Inhibitors

Pandit

2014

Phosphodiesterases and Their Inhibitors

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Identification of PDE4B Over 4D subtype-selective inhibitors revealing an unprecedented binding mode

Cited by 53 publications

References 18 publications

PDE4B mediates local feedback regulation of β1-adrenergic cAMP signaling in a sarcolemmal compartment of cardiac myocytes

PDE4B mediates local feedback regulation of β1-adrenergic cAMP signaling in a sarcolemmal compartment of cardiac myocytes

Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4

PDE4: New Structural Insights into the Regulatory Mechanism and Implications for the Design of Selective Inhibitors

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