This study examines the involvement of cytochromes P-450 and b5 in catalysis of prostaglandin hydroxylation, at the omega- and (omega-1)-positions. To achieve this goal, purified isozymes of cytochromes P-450 from liver and kidney will be utilized. Also, it is planned to isolate the enzyme catalyzing specifically the omega-hydroxylation of prostaglandins. The purification of this enzyme is being attempted by affinity chromatography. If this procedure is not successful, conventional column procedures will be attempted. In an attempt to determine the sequence of metabolic transformation of PGs, it is planned to determine whether beta-oxidation can occur with 19-hydroxy and 20-hydroxy-prostaglandins. Furthermore, the ability of omega-oxidized PGs to undergo omega- or (omega-1)- hydroxylation will be examined.