Assessment of selective inhibition of rat cerebral cortical calcium-independent and calcium-dependent phosphodiesterases in crude extracts using deoxycyclic AMP and potassium ions

Davis, Craig W.

doi:10.1016/0304-4165(84)90257-5

Cited by 74 publications

(41 citation statements)

References 28 publications

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“…Furthermore, the high molecular mass fraction of crude cytosolic preparations from rat brain has been reported to be sensitive to rolipram inhibition [24]. We indeed observed a cyclic-AMP-specific, rolipram-inhibited, 250-kDa enzyme form, by exclusion chromatography of crude brain cytosol on ACA 34 gel.…”

Section: Rolipram-sensitive Phosphodiesterase In Crude Preparationssupporting

confidence: 59%

Isolation of similar rolipram‐inhibitable cyclic‐AMP‐specific phosphodiesterases from rat brain and heart

Némoz

Moueqqit

Prigent

et al. 1989

European Journal of Biochemistry

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A cyclic AMP phosphodiesterase form of rat brain cytosol was purified by means of affinity chromatography on an immobilized analog of the specific inhibitor rolipram, followed by an exclusion chromatography step. The resulting preparation presented two protein bands in polyacrylamide gel electrophoresis, both with phosphodiesterase activity. Kinetics of cyclic AMP hydrolysis by the purified enzyme proved of the Michaclis type, with a K, of 3 pM, while hydrolysis of cyclic GMP displayed anomalous, negatively cooperative kinetics. At micromolar concentrations, this enzyme form hydrolyzed highly specifically cyclic AMP (%fold faster than cyclic GMP). Cyclic GMP proved a poor competitor of cyclic AMP hydrolysis (Ki 1.04 mM). The neurotropic compound, rolipram, strongly inhibited the enzyme, in a competitive manner (Ki 0.9 pM). This enzyme displayed a molecular mass of around 44 kDa as determined by exclusion chromatography, but two molecular masses of 42 kDa and 89 kDa were observable by electrophoresis on a polyacrylamide gradient gel, compatible with an equilibrium between dimeric and monomeric forms. Isoelectric focusing of the preparation gave rise to two activity peaks of PI 4.8 and 6.7, with identical properties, probably representing two charge isomers of the same protein. An enzyme prepared from rat heart cytosol by the same techniques as for brain phosphodiesterase isolation shared numerous characteristics with the enzyme of cerebral origin, suggesting identity of the rolipramsensitive form between the two tissues.Since the rolipram-sensitive form detected in crude brain preparations markedly differs from the abovedescribed isolated enzyme, both by its molecular mass in exclusion chromatography and by its PI, it is suggested that an alteration of the native protein, due to dissociation of putative subunits, occurs during the puriGcation procedure.

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Section: Rolipram-sensitive Phosphodiesterase In Crude Preparationssupporting

confidence: 59%

Isolation of similar rolipram‐inhibitable cyclic‐AMP‐specific phosphodiesterases from rat brain and heart

Némoz

Moueqqit

Prigent

et al. 1989

European Journal of Biochemistry

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“…A number of different PDE isozymes have been described in various tissues which have different substrate specificities and sensitivities to various PDE inhibitors (Davis, 1984;Weishaar et al, 1986;Reeves et al, 1987;Silver et al, 1988). Three PDE activities have been identified in broken cell preparations of tracheal smooth muscle (Torphy et al, 1987;Silver et al, 1988): (1) a calcium-calmodulin stimulated activity (PDE I), which hydrolyses both cyclic AMP and cyclic GMP with similar potencies, and is inhibitable by M & B 22948 (Weishaar et al, 1986;Torphy et al, 1987;Silver et al, 1988); (2) a cyclic AMP-specific activity (PDE III) which is sensitive to i SKF 94120 Reeves et al, 1987) and (3) a rolipram-sensitive activity, specific for cyclic AMP, with a higher Michaelis constant (KM) for the substrate cyclic nucleotide (Torphy et al, 1987;Reeves et al, 1987;Silver et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

Inhibition of histamine‐stimulated inositol phospholipid hydrolysis by agents which increase cyclic AMP levels in bovine tracheal smooth muscle

Hall

Donaldson

Hill

1989

British J Pharmacology

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“…This (10,22,23) and a Km for cAMP of =2 ,uM. This type of enzyme binds to, and is inhibited (10,(23)(24)(25)(26)(27) …”

Section: Discussionmentioning

confidence: 99%

Cloning and characterization of mammalian homologs of the Drosophila dunce+ gene.

Davis

Takayasu²,

Eberwine³

et al. 1989

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

135

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A probe representing the Drosophila dunce'(dnc') gene, the structural gene for a cAMP phosphodiesterase (PDEase), detects homologous sequences in many different organisms, including mouse, rat, and human. Genomic and cDNA clones representing a homolog of the Drosophila dnc' gene were isolated from rat libraries and characterized. This gene has been named ratdnc-1. One cDNA clone defines a large open reading frame of 41.8 kilobases (kb), predicting a protein sequence of 610 amino acids with significant homology to a conserved domain of -275 residues found in most otherPDEases. The amino acid identity value to the Drosophila cAMP PDEase within this domain is a striking 75%. Other cDNA clones show blocks of sequence divergence from this cDNA clone close to the predicted N terminus, indicating the potential existence of a family of related enzymes encoded by alternatively spliced messenger RNAs from ratdnc-1. Genomic blotting experiments suggest the existence of at least one other rat gene with homology to ratdnc-1. RNAs homologous to ratdnc-1 are heterogeneous in size between tissues, with heart containing a major transcript of 4.4 kb and brain one of4.0 kb.The potential identity of the product of the ratdnc-1 gene with known PDEases is discussed.

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Assessment of selective inhibition of rat cerebral cortical calcium-independent and calcium-dependent phosphodiesterases in crude extracts using deoxycyclic AMP and potassium ions

Cited by 74 publications

References 28 publications

Isolation of similar rolipram‐inhibitable cyclic‐AMP‐specific phosphodiesterases from rat brain and heart

Isolation of similar rolipram‐inhibitable cyclic‐AMP‐specific phosphodiesterases from rat brain and heart

Inhibition of histamine‐stimulated inositol phospholipid hydrolysis by agents which increase cyclic AMP levels in bovine tracheal smooth muscle

Cloning and characterization of mammalian homologs of the Drosophila dunce+ gene.

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