Substrate inactivation of lung thromboxane synthase preferentially decreases thromboxane A2 production.
Bovine lung thromboxane synthase was immobilized on phenyl-Sepharose beads by adsorption. The immobilized enzyme was catalytically active and synthesized both TXA2 and HHT. The production of both products was inhibited by 1-benzylimidazole and furegrelate. Multiple additions of PGH2 dramatically reduced the ability of the enzyme to synthesize TXA2, but did not effect the synthesis of HHT. In addition, 1-benzylimidazole did not protect thromboxane synthase from inactivation with multiple additions of PGH2. When the enzyme was incubated with PGH2 in the presence of 1-benzylimidazole, the synthesis of TXA2 was inhibited. When the inhibitor was removed the enzyme had still been inactivated by PGH2 in the presence of 1-benzylimidazole. Thus the substrate inactivation of the enzyme does not require the production of TXA2. Our data suggests that the synthesis of TXA2 and HHT can be differentially inactivated and may occur at different sites on the enzyme.
Hall
E R
ER
Department of Internal Medicine, University of Texas Medical School, Houston.
Townsend
G L
GL
Linthicum
D S
DS
Frasier-Scott
K F
KF
D000818:Animals; D002417:Cattle; D004789:Enzyme Activation; D007093:Imidazoles / Q000494:pharmacology; D008168:Lung / Q000187:drug effects / Q000201:enzymology*; D008861:Microsomes / Q000187:drug effects / Q000201:enzymology; D011450:Prostaglandin Endoperoxides, Synthetic / Q000494:pharmacology; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000494:pharmacology; D013379:Substrate Specificity; D013928:Thromboxane A2 / Q000096:biosynthesis*; D013930:Thromboxane-A Synthase / Q000378:metabolism*
D007093:Imidazoles; D011450:Prostaglandin Endoperoxides, Synthetic; D011463:Prostaglandins H; C017062:1-benzylimidazole; D044262:Prostaglandin H2; D013928:Thromboxane A2; D013930:Thromboxane-A Synthase