An enzymatic method for distinguishing the stereoisomers of 12-hydroxyeicosatetraenoic acid in human epidermis and psoriatic scale.
Homogenates of normal human epidermis synthesized 12-hydroxyeicosatetraenoic acid (12-HETE) when incubated in vitro with arachidonic acid. The stereoconfigurations of the C-12 hydroxyl isomers were determined by incubation with potato 5-lipoxygenase. The synthesized substrate-specific diHETEs; 5S, 12R and 5S, 12S, were readily separated by high performance liquid chromatography. Using this novel methodology, the normal epidermis was found to synthesize predominantly 12-S-HETE while, in contrast, psoriatic scale was found to contain 12-R-HETE. The 5-lipoxygenase inhibitors, Merck L-651, 896, Takeda AA86I, and the active metabolite of Syntex lonapalene were found to inhibit 12-HETE formation in normal epidermal homogenates.
Opas
E E
EE
Department of Biochemistry and Molecular Biology, Merck Sharp and Dohme Research Laboratories, Rahway, New Jersey 07065.
Argenbright
L W
LW
Humes
J L
JL
D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004817:Epidermis / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D006893:Hydroxyeicosatetraenoic Acids / Q000096:biosynthesis*; D008084:Lipoxygenase / Q000378:metabolism; D016859:Lipoxygenase Inhibitors; D008722:Methods; D009249:NADP / Q000494:pharmacology; D010640:Phenothiazines / Q000494:pharmacology; D011565:Psoriasis / Q000201:enzymology / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D013237:Stereoisomerism
D001095:Arachidonic Acids; D006893:Hydroxyeicosatetraenoic Acids; D016859:Lipoxygenase Inhibitors; D010640:Phenothiazines; D011725:Pyridines; C013109:metapyrone; D016718:Arachidonic Acid; D009249:NADP; D019377:12-Hydroxy-5,8,10,14-eicosatetraenoic Acid; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; C053726:4-bromo-2,7-dimethoxy-3H-phenothiazin-3-one; D008084:Lipoxygenase