The upstream conserved regions (UCRs) mediate homo- and hetero-oligomerization of type 4 cyclic nucleotide phosphodiesterases (PDE4s)

Xie, Moses; Blackman, Brigitte E.; Scheitrum, Colleen; Mika, Delphine; Blanchard, Elise; Tao, Lei; Conti, Marco; Richter, Wito

doi:10.1042/bj20131681

Cited by 29 publications

(31 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Two pivotal actions have been evidenced for UCR1 and UCR2 domains. Firstly, they promote the long forms dimerization in vivo , involving particularly the UCR1/catalytic domain interfaces . On the other hand, UCR2, closing across the active site, blocks the access to cAMP, thus exerting a partial enzyme inhibition ,…”

Section: Introductionmentioning

confidence: 99%

Further Insights in the Binding Mode of Selective Inhibitors to Human PDE4D Enzyme Combining Docking and Molecular Dynamics

D’Ursi

Guariento

Trombetti

et al. 2016

Molecular Informatics

View full text Add to dashboard Cite

Alzheimer′s disease has recently emerged as a possible field of application for PDE4D inhibitors (PDE4DIs). The great structure similarity among the various PDE4 isoforms and, furthermore, the lack of the full length crystal structure of the enzyme, impaired the rational design of new selective PDE4DIs. In this paper, with the aim of exploring new insights into the PDE4D binding, we tackled the problem by performing a computational study based on docking simulations combined with molecular dynamics (D‐MD). Our work uniquely identified the binding mode and the key residues involved in the interaction with a number of in‐house catechol iminoether derivatives, acting as PDE4DIs. Moreover, the new binding mode was tested using a series of analogues previously reported by us and it was used to confirm their key structural features to allow PDE4D inhibition. The binding model disclosed within the current computational study may prove to be useful to further advance the design and synthesis of novel, more potent and selective, PDE4D inhibitors.

show abstract

Section: Introductionmentioning

confidence: 99%

Further Insights in the Binding Mode of Selective Inhibitors to Human PDE4D Enzyme Combining Docking and Molecular Dynamics

D’Ursi

Guariento

Trombetti

et al. 2016

Molecular Informatics

View full text Add to dashboard Cite

show abstract

“…These include the Ca 2 + /calmodulin binding domains of PDE1, the Gaf domains of PDE2, PDE5, PDE6, PDE10, PDE11 and the UCR1/2 domains of PDE4 [2,9,15,17] . Such domains have been implicated in dimer formation [35–46] .…”

Section: Introductionmentioning

confidence: 99%

Dimerization of cAMP phosphodiesterase-4 (PDE4) in living cells requires interfaces located in both the UCR1 and catalytic unit domains

Bolger

Dunlop

Meng

et al. 2015

Cellular Signalling

View full text Add to dashboard Cite

PDE4 family cAMP phosphodiesterases play a pivotal role in determining compartmentalised cAMP signalling through targeted cAMP breakdown. Expressing the widely found PDE4D5 isoform, as both bait and prey in a yeast 2-hybrid system, we demonstrated interaction consistent with the notion that long PDE4 isoforms form dimers. Four potential dimerization sites were uncovered using a scanning peptide array approach, where a recombinant purified PDE4D5 fusion protein was used to probe a 25-mer library of overlapping peptides covering the entire PDE4D5 sequence. Key residues involved in PDE4D5 dimerization were defined using a site-directed mutagenesis programme directed by an alanine scanning peptide array approach. Critical residues stabilising PDE4D5 dimerization were defined within the regulatory UCR1 region found in long, but not short, PDE4 isoforms, namely the Arg173, Asn174 and Asn175 (DD1) cluster. Disruption of the DD1 cluster was not sufficient, in itself, to destabilise PDE4D5 homodimers. Instead, disruption of an additional interface, located on the PDE4 catalytic unit, was also required to convert PDE4D5 into a monomeric form. This second dimerization site on the conserved PDE4 catalytic unit is dependent upon a critical ion pair interaction. This involves Asp463 and Arg499 in PDE4D5, which interact in a trans fashion involving the two PDE4D5 molecules participating in the homodimer. PDE4 long isoforms adopt a dimeric state in living cells that is underpinned by two key contributory interactions, one involving the UCR modules and one involving an interface on the core catalytic domain. We propose that short forms do not adopt a dimeric configuration because, in the absence of the UCR1 module, residual engagement of the remaining core catalytic domain interface provides insufficient free energy to drive dimerization. The functioning of PDE4 long and short forms is thus poised to be inherently distinct due to this difference in quaternary structure.

show abstract

“…Phosphorylation by protein kinase A (PKA) at a conserved site on UCR1 activates all long PDE4 isoforms (8,9). Mutation and deletion studies have shown that long forms of PDE4 are dimeric, with key dimerization interactions mediated by UCR1 and UCR2 (10,11), and that the C-terminal half of UCR2 could play a negative regulatory role (12)(13)(14)(15)(16). Access to details of these different mechanisms of control would advance our capacity to design medicines that act selectively among the PDE4 isoforms, and should also cast light on their specific intracellular functions.…”

mentioning

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.

View full text Add to dashboard Cite

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...

show abstract

The upstream conserved regions (UCRs) mediate homo- and hetero-oligomerization of type 4 cyclic nucleotide phosphodiesterases (PDE4s)

Cited by 29 publications

References 41 publications

Further Insights in the Binding Mode of Selective Inhibitors to Human PDE4D Enzyme Combining Docking and Molecular Dynamics

Further Insights in the Binding Mode of Selective Inhibitors to Human PDE4D Enzyme Combining Docking and Molecular Dynamics

Dimerization of cAMP phosphodiesterase-4 (PDE4) in living cells requires interfaces located in both the UCR1 and catalytic unit domains

Engineered stabilization and structural analysis of the autoinhibited conformation of PDE4

Contact Info

Product

Resources

About