Human and mouse eLOX3 have distinct substrate specificities: implications for their linkage with lipoxygenases in skin.
Genetic and biochemical evidence suggests a functional link between human 12R-lipoxygenase (12R-LOX) and epidermal lipoxygenase-3 (eLOX3) in normal differentiation of the epidermis; LOX-derived fatty acid hydroperoxide is isomerized by the atypical eLOX3 into a specific epoxyalcohol that is a potential mediator in the pathway. Mouse epidermis expresses a different complement of LOX enzymes, and therefore this metabolic linkage could differ. To test this concept, we compared the substrate specificities of recombinant mouse and human eLOX3 toward sixteen hydroperoxy stereoisomers of arachidonic and linoleic acids. Both enzymes metabolized R-hydroperoxides 2-3 times faster than the corresponding S enantiomers. Whereas 12R-hydroperoxyeicosatetraenoic acid (12R-HPETE) is the best substrate for human eLOX3 (2.4 s(-1); at 30 microM substrate), mouse eLOX3 shows the highest turnover with 8R-HPETE (2.9 s(-1)) followed by 8S-HPETE (1.3 s(-1)). Novel product structures were characterized from reactions of mouse eLOX3 with 5S-, 8R-, and 8S-HPETEs. 8S-HPETE is converted specifically to a single epoxyalcohol, identified as 10R-hydroxy-8S,9S-epoxyeicosa-5Z,11Z,14Z-trienoic acid. The substrate preference of mouse eLOX3 and the unique occurrence of an 8S-LOX enzyme in mouse skin point to a potential LOX pathway for the production of epoxyalcohol in murine epidermal differentiation.
Yu
Zheyong
Z
Division of Clinical Pharmacology, Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Schneider
Claus
C
Boeglin
William E
WE
Brash
Alan R
AR
D000818:Animals; D001092:Arachidonate 12-Lipoxygenase / Q000737:chemistry*; D001095:Arachidonic Acids / Q000737:chemistry*; D004789:Enzyme Activation; D006801:Humans; D008041:Linoleic Acids / Q000737:chemistry*; D008084:Lipoxygenase / Q000737:chemistry*; D051379:Mice; D012867:Skin / Q000201:enzymology*; D013045:Species Specificity
D001095:Arachidonic Acids; D008041:Linoleic Acids; C095864:lipoxygenase 3; D008084:Lipoxygenase; D001092:Arachidonate 12-Lipoxygenase