Cyclic AMP metabolism by swine adipocyte microsomal and plasma membranes.
Extracellular cyclic AMP is source of extracellular adenosine in brain and kidney. Whether this occurs in adipose tissue is unknown. The present study evaluated the capacity of swine adipocyte plasma membranes to metabolize cyclic AMP to AMP and adenosine, via phosphodiesterase (PDE) and 5'-nucleotidase (5'-NT), respectively. Plasma membranes (PM) and microsomal membranes (MM) were isolated from over-the-shoulder subcutaneous adipose tissue of 3 month-old male miniature swine. The purity of the membrane fractions was determined and PDE and 5'-NT activities in PM and MM fractions were corrected for cross-contamination. The maximal activity of MM-PDE was 7-fold greater than that of PM-PDE. MM-PDE was 100% inhibited by 5 microM cilostamide, while PM-PDE was unaffected by this PDE3B inhibitor. Inhibitors of PDE1, PDE2, PDE4 and PDE5 also failed to inhibit PM-PDE. However, 1 mM DPSPX inhibited PM-PDE activity by 72%. When PM were incubated with 0.8 microM cyclic AMP for 20 min, AMP accumulation was four times that of adenosine. These data demonstrate that cyclic AMP can be converted to AMP and adenosine by the PM-bound enzymes 5'-NT and PDE, and suggest that the PM-PDE responsible for extracellular cyclic AMP metabolism to AMP is distinct from the intracellular MM-PDE.
Zacher
L A
LA
Department of Animal and Nutritional Sciences, University of New Hampshire, Durham 03824, USA.
Carey
G B
GB
D015720:5'-Nucleotidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D000241:Adenosine / Q000378:metabolism; D000249:Adenosine Monophosphate; D017667:Adipocytes / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D000818:Animals; D015415:Biomarkers; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D007425:Intracellular Membranes / Q000201:enzymology / Q000378:metabolism; D007700:Kinetics; D008565:Membrane Proteins / Q000737:chemistry; D008861:Microsomes / Q000201:enzymology / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000378:metabolism; D020033:Protein Isoforms / Q000032:analysis; D058915:Purinergic P1 Receptor Antagonists; D015363:Quinolones / Q000494:pharmacology; D015290:Second Messenger Systems; D013552:Swine; D014970:Xanthines / Q000494:pharmacology
D015415:Biomarkers; D008565:Membrane Proteins; D010726:Phosphodiesterase Inhibitors; D020033:Protein Isoforms; D058915:Purinergic P1 Receptor Antagonists; D015363:Quinolones; D014970:Xanthines; D000249:Adenosine Monophosphate; C021294:cilostamide; C053896:1,3-dipropyl-8-(4-sulfophenyl)xanthine; D000242:Cyclic AMP; D015720:5'-Nucleotidase; D010727:Phosphoric Diester Hydrolases; D000241:Adenosine