Platelet cyclic adenosine monophosphate phosphodiesterases: targets for regulating platelet-related thrombosis.
Platelets contain two cyclic adenosine monophosphate (cAMP) phosphodiesterases (PDEs) that regulate the level of cAMP, the major inhibitor of platelet activation pathways. PDE3A hydrolyzes cAMP to 5' AMP with a low K (m). PDE3A is inhibited by cyclic guanosine monophosphate (cGMP), which provides a feedback control and controls basal levels of cAMP. In contrast, PDE2A hydrolyzes both cAMP and cGMP with a high K (m), is allosterically stimulated by cGMP at moderate levels, and may control the stimulated levels of cAMP. Using affinity labeling, chemical modification, and site-directed mutagenesis of highly conserved amino acids, the amino acids required for catalytic activity and/or metal binding are H752 and H756. The singular binding sites for cAMP include N845, E971, and F972, whereas the unique amino acids interacting with cGMP are Y751, H836, H849, and D950. Residues E866 and F1004 are present in both the overlapping cGMP and cAMP sites. Two inhibitors of PDE3A are used in clinical medicine: milrinone and cilostazol. Three amino acids, Y751, D950, and F1004, show decreased sensitivity to both inhibitors (increased K (i)). These inhibitors mimic cGMP as an inhibitor of PDE3A rather than compete for cAMP binding. New nonhydrolyzable affinity labels inactivate PDE3A and are protected by Sp-cAMPS, a nonhydrolyzable substrate of the enzyme. These compounds have the potential to identify amino acids that are unique for PDE3A. An inhibitor of platelet PDE2A increases cAMP more than inhibitors of PDE3A but has much less effect on platelet activation, suggesting that these enzymes are present in different compartments of the cell.
Colman
Robert W
RW
The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, Pennsylvania 19140, USA. colmanr@astro.temple.edu
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000097:blood* / Q000235:genetics; D000818:Animals; D001792:Blood Platelets / Q000502:physiology*; D017124:Conserved Sequence; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; D006801:Humans; D008954:Models, Biological; D015539:Platelet Activation; D013927:Thrombosis / Q000097:blood*
D015105:3',5'-Cyclic-AMP Phosphodiesterases; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; C517287:PDE3A protein, human