Cytosolic and sarcoplasmic reticulum-associated low Km, cGMP-inhibited cAMP phosphodiesterase in mammalian myocardium.
The distribution and phosphoprotein band patterns of low Km, cGMP-inhibited cAMP phosphodiesterase (cGI PDE) activity were examined in cytosolic and microsomal fractions of human, canine, rabbit and guinea pig left ventricular myocardium following phosphorylation by cAMP-dependent protein kinase, immunoprecipitation with anti-cGI PDE antibodies and SDS-PAGE. The recovery of cGI PDE activity in cytosolic and microsomal fractions was comparable in all four species. Microsomal cGI PDE was comprised chiefly of a approximately 135 kDa phosphoprotein. Cytosolic cGI PDE was comprised solely of approximately 116 kDa and lower molecular weight phosphoproteins. The approximately 135 kDa phosphoprotein probably corresponds to the holoenzyme encoded by the recently cloned cDNA for human myocardial cGI PDE, whose predicted molecular weight is 126 kDa. The approximately 116 kDa phosphoprotein may result from deletion or removal of putative membrane-association domains from the N-terminal region of the holoenzyme. These results suggest that the cytosolic and sarcoplasmic reticulum-associated forms of mammalian myocardial cGI PDE are separate molecular species.
Laboratory of Cellular Metabolism, National Heart, Lung & Blood Institute, Bethesda, Maryland.
C J
CJ
Smith
J
J
Krall
V C
VC
Manganiello
M A
MA
Movsesian
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D000242:Cyclic AMP / Q000494:pharmacology; D006152:Cyclic GMP / Q000494:pharmacology*; D003600:Cytosol / Q000201:enzymology; D004285:Dogs; D006168:Guinea Pigs; D006801:Humans; D007163:Immunosorbent Techniques; D008861:Microsomes / Q000201:enzymology; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology* / Q000648:ultrastructure; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011817:Rabbits; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*
D010750:Phosphoproteins; D000242:Cyclic AMP; D011494:Protein Kinases; D015105:3',5'-Cyclic-AMP Phosphodiesterases; D006152:Cyclic GMP