Purification and characterization of high-affinity cyclic adenosine monophosphate phosphodiesterase from dog kidney

Thompson, W. J.; Epstein, Paul M.; Strada, Samuel J.

doi:10.1021/bi00590a030

Cited by 83 publications

(28 citation statements)

References 49 publications

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“…This allowed for the categorization of various cyclic AMPspecific PDE (4) species that had been isolated and purified much earlier (11)(12)(13). Substantiation of the PDE4 enzyme family was achieved definitively by homology cloning, initially, using the Drosophila dunc PDE gene as a probe to isolate rat PDE4 species (14,15).…”

mentioning

confidence: 99%

The Human Cyclic AMP-specific Phosphodiesterase PDE-46 (HSPDE4A4B) Expressed in Transfected COS7 Cells Occurs as Both Particulate and Cytosolic Species That Exhibit Distinct Kinetics of Inhibition by the Antidepressant Rolipram

Huston

Pooley

Julien

et al. 1996

Journal of Biological Chemistry

152

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Transfection of COS7 cells with a plasmid encoding the human cyclic AMP-specific PDE4A phosphodiesterase PDE-46 (HSPDE4A4B) led to the expression of a rolipram-inhibited PDE4 activity, which contributed ϳ96% of the total COS cell PDE activity. A fusion protein was generated which encompassed residues (788 -886) at the extreme C terminus of PDE-46 and was used to generate an antiserum that detected PDE-46 in transfected COS7 cells. Immunoblotting studies identified PDE-46 as a ϳ125-kDa species that was associated with both the soluble and particulate fractions. The relative V max of particulate PDE-46 was ϳ56% that of cytosolic PDE-46. Particulate PDE-46 was not solubilized using Triton X-100 or high NaCl concentrations. Immunofluorescence analysis by laser scanning confocal microscopy showed that PDE-46 was located at discrete margins of the cell, indicative of association with membrane cortical regions. The human PDE4A species, h6.1 (HSPDE4A4C), which lacks the N-terminal extension of PDE-46, was found as an entirely soluble species when expressed in COS7 cells. h6.1 was shown to have an ϳ11-fold higher V max relative to that of PDE-46. In doseresponse studies rolipram inhibited particulate PDE-46 at much lower concentrations (IC 50 ‫؍‬ 0.195 M) than those needed to inhibit the cytosolic enzyme (IC 50 ‫؍‬ 1.6 M). The basis of this difference lay in the fact that rolipram served as a simple competitive inhibitor of the cytosol enzyme (K i ‫؍‬ 1.6 M) but as a partial competitive inhibitor of the particulate enzyme (K i ‫؍‬ 0.037 M; K i ‫؍‬ 2.3 M). Particulate PDE-46 thus showed a ϳ60-fold higher affinity for rolipram than cytosolic PDE-46.

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mentioning

confidence: 99%

The Human Cyclic AMP-specific Phosphodiesterase PDE-46 (HSPDE4A4B) Expressed in Transfected COS7 Cells Occurs as Both Particulate and Cytosolic Species That Exhibit Distinct Kinetics of Inhibition by the Antidepressant Rolipram

Huston

Pooley

Julien

et al. 1996

Journal of Biological Chemistry

152

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“…2) Effect of amrinone on cAMPPDE activity: cAMPPDE of the homogenates of the smooth muscle cells were partially purified by DEAF cellulose chromatography (19,20). The cells grown to confluence in a 75 cm2 flask (Corning) were harvested by a rubber policeman, resuspended in homo genizing buffer (10 mM Tris/CI, pH 8.0; 50 mM NaF, 20 ,uM N-p-tosyl-L-lysine chloromethyl ketone (TLCK); 20 uM leupeptin and 1% Triton x-100 (V/V)) and homogenized in a glass homogenizer with a Teflon pestle using 10 strokes by hand at 0°C.…”

Section: Methodsmentioning

confidence: 99%

Involvement of Cyclic AMP in Vasodilatation by Amrinone: A Comparative Study with 3-Isobutyl-1-Methyl-Xanthine (IBMX)

Kishi

Numano

1986

Japanese Journal of Pharmacology

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Abstract-We investigated the effect of amrinone, a non-glycosidic, non-adrenergic cardiotonic drug, on rabbit aortic strips and vascular smooth muscle cells cultured from rabbit aorta. Amrinone relaxed the strips precontracted by KCI, norepinephrine, serotonin or STA2 in a dose-dependent manner, and it shifted the dose-response curves downward.A similar mode of relaxation was noted with IBMX, an inhibitor of cyclic AMP phosphodiesterase (cAMPPDE). Although propranolol did not affect the relaxation induced by amrinone, W-7, a calmodulin antagonist, slightly potentiated and IBMX attenuated the response.In intact smooth muscle cells in culture, amrinone increased basal levels of cAMP and markedly potentiated cAMP accumulation in response to 10-6 M isoproterenol. The effect on cAMP accumu lation mimicked that of IBMX but the effects were not additive.Inhibition of cAMPPDE was also demonstrated in a cell-free system, IC50 values for amrinone and IBMX being 2

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“…This PDE family is widely distributed in several tissues and has been studied in details in mammalian liver [26]. kidney [27], testis [28] and brain [29]. In all these tissues a certain degree of hydrophobicity and molecular asymmetry, resulting in a particularly high frictional ratio.…”

Section: Neuroblastomamentioning

confidence: 99%

Characterization of 3':5' cyclic nucleotide phosphodiesterase activities of mouse neuroblastoma N18TG2 cells

et al. 1993

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nucleottde phosphodtesterase acttvities (PDE) expressed m mouse N 18TG3 neuroblastoma cells IS reported. At least 3 peaks of acttvity were isolated by DEAE chromatography.none of which was calcmm-calmodulin sttmulated and cGMP sttmulated or mhibtted A first peak elutes at 200 mM sodium acetate: it specifically hydrolyzes cGMP with a II',;, of 4.7 PM and shows senstttvity to zaprinast [M&B 129481 (I .8 PM). A second peak elutmg at 410 mM sodturn acetate hydrolyzes both cychc nucleotides. A third peak. spectfic for CAMP hydrolyses. elutes at 580 mM sodtum acetate. has a K,,,, of 3.1 PM and IS senstttve to RO 20 1723 (7.6 PM) and rohpram (2 /M ). Hydrodynamtc analysts showed for the first peak a Stokes radius of 5.3 nm wtth a sedtmentatton coeffictent of 8 I S. a frtctional ratto tJ7f,,) of I .3l and a nattve molecular mass of IS? kDa. The same analysts for peak 3 showed a Stokea radius of 4 I nm wtth a sedimentatton coeffictent of 3 7 S. a fractional ratto of I 63 and a native molecular mass of 56 kDa The btochemtcal features reported for the enzyme eluttng m the first peak. and Its cGMP-bmding acttvity sttmulated by mhibttors of phosphodtesterase acttvtty. demonstrate that tt belongs to the PDE V subfamily; on the other hand the cAMP sprc~fic enzyme eluting m the third peak can be assigned to the 'RO 20 1724 inhibtted' form. The sigmficance of these findings is dtscussed m relatton to the functtonal characteristic5 of the NIRTG:! cell line Hugh affimty cychc nucleottde phosphodiesterase: PDE V subfamtly: Neuroblastoma cell: Imtdarolinone: Zaprmast

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Purification and characterization of high-affinity cyclic adenosine monophosphate phosphodiesterase from dog kidney

Cited by 83 publications

References 49 publications

The Human Cyclic AMP-specific Phosphodiesterase PDE-46 (HSPDE4A4B) Expressed in Transfected COS7 Cells Occurs as Both Particulate and Cytosolic Species That Exhibit Distinct Kinetics of Inhibition by the Antidepressant Rolipram

The Human Cyclic AMP-specific Phosphodiesterase PDE-46 (HSPDE4A4B) Expressed in Transfected COS7 Cells Occurs as Both Particulate and Cytosolic Species That Exhibit Distinct Kinetics of Inhibition by the Antidepressant Rolipram

Involvement of Cyclic AMP in Vasodilatation by Amrinone: A Comparative Study with 3-Isobutyl-1-Methyl-Xanthine (IBMX)

Characterization of 3':5' cyclic nucleotide phosphodiesterase activities of mouse neuroblastoma N18TG2 cells

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