Figure 4.

HPLC analysis of hydroxy fatty acids formed from LA by the lipid body LOX and its H608V mutant. Equal amounts of LOX protein were incubated with 0.9 mM LA at room temperature for 30 min. After reduction of lipids with sodium borohydride, the reaction mixture was acidified and the lipids were extracted. Oxygenated fatty acid derivatives were isolated by RP-HPLC, and positional isomers were analyzed by SP-HPLC. Ratios of S and R were determined by CP-HPLC (Insets).