Oxidation and keto reduction of 12-hydroxy-5,8,10,14-eicosatetraenoic acids in bovine corneal epithelial microsomes.
The R and S enantiomers of 12-hydroxyeicosatetraenoic acid (12-HETE) exhibit different biological activities. Although they appear to be produced by different enzymatic pathways, cytochrome P-450 monooxygenase and lipoxygenase, respectively, they display similar metabolism in both corneal epithelium and neutrophils. In corneal epithelial microsomes, both enantiomers are subject to oxidation and keto reduction reactions to form the dihydro metabolite, 12-hydroxy-5,8,14-eicosatrienoic acid (12-HETrE), via a keto intermediate. The apparent Km for the formation of 12-HETrE was 17.9 and 20 microM for 12(R)-HETE and 12(S)-HETE, respectively, and the apparent Vmax of the reaction was 17.4 and 8.2 pmol/mg per min, respectively. Chiral analysis of the dihydro metabolite demonstrated a product enantiospecificity. Arachidonic acid, 12(R)-HETE, 12(S)-HETE and the intermediate of this reaction, 12-oxo-ETrE, were metabolized predominantly to 12(R)-HETrE in a ratio [12(R)-HETrE: 12(S)-HETrE] of 7.3:1, 4.3:1, 1.5:1 and 2.3:1, respectively. 12(R)-HETrE is a potent vasodilator, chemotactic and angiogenic factor whose synthesis is induced in inflamed tissues; 12(S)HETrE is devoid of these properties. 12(R)-HETE, derived from NADPH-dependent cytochrome P-450 monooxygenases, and 12(S)-HETE, derived from 12-lipoxygenase, may both play an important role in regulating the inflammatory response by serving as substrates for the local synthesis of 12(R)-HETrE.
Yamamoto
S
S
Department of Pharmacology, New York Medical College, Valhalla, NY 10595.
Nishimura
M
M
Conners
M S
MS
Stoltz
R A
RA
Falck
J R
JR
Chauhan
K
K
Laniado-Schwartzman
M
M
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D015126:8,11,14-Eicosatrienoic Acid / Q000031:analogs & derivatives* / Q000378:metabolism; D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism; D002417:Cattle; D003315:Cornea / Q000378:metabolism* / Q000648:ultrastructure; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism; D006893:Hydroxyeicosatetraenoic Acids / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D009249:NADP; D010084:Oxidation-Reduction; D013237:Stereoisomerism
D006893:Hydroxyeicosatetraenoic Acids; C067514:12-hydroxy-5,8,14-eicosatrienoic acid; D016718:Arachidonic Acid; D009249:NADP; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D003577:Cytochrome P-450 Enzyme System; D001092:Arachidonate 12-Lipoxygenase; D015126:8,11,14-Eicosatrienoic Acid