Table 3.
Species (mutant) | Relative product share, % | |
15-HETE | 12-HETE | |
Human (WT) | 78.7 ± 1.6 | 21.3 ± 1.6 |
Human (Met419Thr) | 34.7 ± 8.7 | 65.3 ± 8.7 |
Human (Thr594Val) | 81.5 ± 1.2 | 18.6 ± 0.9 |
Gibbon (WT) | 46.1 ± 5.2 | 53.9 ± 5.2 |
Gibbon (Ile418Ala) | <1 | 99 ± 14 |
Gibbon (Ile418Phe) | 83.6 ± 3.5 | 16.4 ± 3.5 |
Human and gibbon ALOX15 orthologs were expressed in E. coli. The product pattern was analyzed by consecutive RP-HPLC, SP-HPLC, and chiral-phase HPLC. Four independent expression experiments were performed, and each sample was quantified twice. Means ± SD of the relative shares of the two major reaction products are given.