[43] Cytochrome P-450 arachidonate oxygenase
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2007, Archives of Biochemistry and BiophysicsCitation Excerpt :Subcellular fractions were stored at −80 °C. Arachidonic acid metabolism was assayed by the method of Capdevila et al. [10,28]. Reaction mixtures (0.25 ml total volume) contained 75–150 μg of protein as indicated in the figure legends, and 30 μM [1-14C] arachidonic acid (53 mCi/mmol) (Perkin Elmer, Torrance, CA).
Peroxisomal proliferator-activated receptor-α-dependent inhibition of endothelial cell proliferation and tumorigenesis
2007, Journal of Biological ChemistryCitation Excerpt :Averaged tumor volumes were calculated using the following formula: tumor volume (mm3) = (length × width2)/2 (37). Western Blot and Immunofluorescence Analyses—Microsomal fractions from mouse liver, temperature-sensitive endothelial, or p60.5 cells were isolated by differential centrifugation as described (38). Microsomal proteins (20–50 μg of protein/well) were resolved by SDS-PAGE in 10% gels and transferred to Immobilon-P membranes (Millipore), and the membranes were incubated with an anti-Cyp2c44 peptide antibody raised against the IGRHQPPSMKDKMKC peptide (GenScript) (39) or rat anti-CYP2C11 antibody (cross-reactive toward mouse Cyp2c29, -2c38, and -2c40, >70% amino acid homology) (40).
P450 CYP2C epoxygenase and CYP4A ω-hydroxylase mediate ciprofibrate-induced PPARα-dependent peroxisomal proliferation
2007, Journal of Lipid ResearchCitation Excerpt :For both incubation times, radioactivity was recovered mainly as [1-14C]AA and also in metabolites eluting earlier than EETs (peak X in Fig. 3A–C). Radioactivity in peak X, which presents a retention time corresponding roughly to that of ω/ω-1 hydroxy acids, such as HEETs (26), increased 10–15% in ciprofibrate-treated cells. However, ω/ω-1 hydroxylation is one of the known routes for EET metabolism (45), and peak X probably contains other oxidized metabolites of AA.