Characterization of an in vivo hormonally regulated phosphodiesterase 3 (PDE3) associated with a liver Golgi-endosomal fraction.
The biochemical properties of an in vivo hormonally regulated low Km cAMP phosphodiesterase (PDE) activity associated with a liver Golgi-endosomal (GE) fraction have been characterized. DEAE-Sephacel chromatography of a GE fraction solubilized by a lysosomal extract resulted in the sequential elution of three peaks of activity (numbered I, II, and III), while ion-exchange HPLC resolved five peaks of activity (numbered 1, 2, 3, 4, and 5). Based on the sensitivity of the eluted activity to cGMP and selected phosphodiesterase inhibitors, two phosphodiesterase isoforms were resolved: a cGMP-stimulated and EHNA-inhibited PDE2, eluted in DEAE-Sephacel peak I and HPLC peak 2 and a cGMP-, a cilostamide-, and ICI 118233-inhibited PDE3, eluted in DEAE-Sephacel peak III and HPLC peaks 3, 4, and 5. GE fractions isolated after acute treatments with insulin, tetraiodoglucagon, and growth hormone displayed an increase in phosphodiesterase activity relative to saline-injected controls, as did GE fractions from genetically obese and hyperinsulinemic rats relative to lean littermates. In all experimental rats, an increase in PDE3 activity associated with DEAE-Sephacel peak III and HPLC peaks 4 and 5 was observed relative to control animals. Furthermore, in genetically obese Zucker rats, an increase in the sensitivity of PDE activity to cilostamide and in the amount of PDE activity immunoprecipitated by an antibody to adipose tissue PDE3 was observed relative to lean littermates. These results extend earlier studies on isolated hepatocytes and show that liver PDE3 is the main if not sole PDE isoform activated by insulin, glucagon, and growth hormone in vivo.
Geoffroy
V
V
INSERM U530, Groupe Hospitalier Necker, Enfants-Malades, Paris, France.
Fouque
F
F
Lugnier
C
C
Desbuquois
B
B
Benelli
C
C
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D006152:Cyclic GMP / Q000494:pharmacology; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; D011992:Endosomes / Q000201:enzymology*; D015971:Gene Expression Regulation, Enzymologic; D005934:Glucagon / Q000031:analogs & derivatives; D006056:Golgi Apparatus / Q000201:enzymology*; D013006:Growth Hormone / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008297:Male; D009765:Obesity; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D011724:Pyridazines; D015363:Quinolones; D051381:Rats; D011924:Rats, Zucker; D013347:Subcellular Fractions / Q000201:enzymology
D006728:Hormones; D007328:Insulin; D007527:Isoenzymes; D010726:Phosphodiesterase Inhibitors; D011724:Pyridazines; D015363:Quinolones; C021294:cilostamide; D013006:Growth Hormone; D005934:Glucagon; C054775:ICI 118233; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; D006152:Cyclic GMP