Molecular cloning and characterization of a distinct human phosphodiesterase gene family: PDE11A

Abstract

We report here the cloning, expression, and characterization of human PDE11A1, a member of a distinct cyclic nucleotide phosphodiesterase (PDE) family. PDE11A exhibits <50% amino acid identity with the catalytic domains of all other PDEs, being most similar to PDE5, and has distinct biochemical properties. The human PDE11A1 cDNA isolated contains a complete open reading frame encoding a 490-amino acid enzyme with a predicted molecular mass of 55,786 Da. At the N terminus PDE11A1 has a single GAF domain homologous to that found in other signaling molecules, including PDE2, PDE5, PDE6, and PDE10, which constitutes a potential allosteric binding site for cGMP or another small ligand. Tissue distribution studies indicate that PDE11A mRNA occurs at highest levels in skeletal muscle, prostate, kidney, liver, pituitary, and salivary glands and testis. PDE11A is expressed as at least three major transcripts of Ϸ10.5, Ϸ8.5, and Ϸ6.0 kb, thus suggesting the existence of multiple subtypes. This possibility is further supported by the detection of three distinct proteins of Ϸ78, Ϸ65, and Ϸ56 kDa by Western blotting of human tissues for PDE11A isoforms. Recombinant human PDE11A1 hydrolyzes both cGMP and cAMP with K m values of 0.52 M and 1.04 M, respectively, and similar V max values. Therefore, PDE11A represents a dual-substrate PDE that may regulate both cGMP and cAMP under physiological conditions. PDE11A is sensitive to the nonselective PDE inhibitor 3-isobutyl-1-methylxanthine (IBMX) as well as zaprinast and dipyridamole, inhibitors that are generally considered relatively specific for the cGMP-selective PDEs, with IC50 values of 49.8 M, 12.0 M, and 0.37 M, respectively.

Prostaglandin E2 Inhibits NLRP3 Inflammasome Activation through EP4 Receptor and Intracellular Cyclic AMP in Human Macrophages

“…To determine, if this is the major mechanism of the observed effect of PGE 2 -induced NLRP3 inhibition in human primary MDM, we applied several approaches that involved the key molecules in the regulation of intracellular cAMP levels. We used KH7, an adenylate cyclase inhibitor which decreases basal or activated cAMP levels (49); forskolin, a direct adenylate cyclase activator, that increases intracellular cAMP levels (50) and IBMX, a phosphodiesterase inhibitor, that increases cAMP levels by blocking its degradation (51). First, we confirmed that in human primary MDM, 15 min treatment with PGE 2 stimulated cAMP production (Fig.…”

Expression of guanylyl cyclase-B (GC-B/NPR2) receptors in normal human fetal pituitaries and human pituitary adenomas implicates a role for C-type natriuretic peptide

“…Although the presence of a pituitary natriuretic peptide system in other vertebrate species suggests that cGMP signaling is important for normal pituitary function, understanding of whether a Endocrine-Related Cancer (2012) 19 497-508 www.endocrinology-journals.org similar system is present in human pituitaries is very limited. Before our current observations of pituitary NPPC and NPR2 expression, previous studies on humans suggest that normal pituitary tissue expresses NPPB (encoding BNP), PRKG2 (encoding protein kinase G II), and PDE11A (encoding phosphodiesterase 11A) (Gerbes et al 1994, Fawcett et al 2000, Zhan & Desiderio 2004. The presence of both PKG2 and PDE11A in human pituitaries indicates potential cGMP-target proteins, which might regulate the effects of CNP/GC-B signaling.…”

T cell activation up-regulates cyclic nucleotide phosphodiesterases 8A1 and 7A3

“…Currently, 11 major families of PDEs (PDEs 1-11), subdivided on the basis of substrate specificity, kinetic properties, inhibitor profiles, and sequence homology (9)(10)(11)(12)(13), have been identified. PDEs 3B, 4A, 4B, 4D, and 7A1 (14)(15)(16) are reported in human T cells.…”