Reconstruction of aryl hydrocarbon hydroxylase from rabbit intestinal mucosa microsomes.
Benzo(a)pyene hydroxylation activity was solubilized from rabbit intestinal mucosa microsomes and reconstituted with a cytochrome P-450 preparation obtained by fractionation with 6-amino-n-hexyl Sepharose 4B, hydroxylapatite and CM-Sephadex C-50, and partially purified NADPH-cytochrome c reductase. Phosphatidylserine was required for the maximal activity, while phosphatidylcholine had no stimulatory effect. The carbon monoxide difference spectrum of the cytochrome P-450 fraction showed a maximum peak at 450 nm. Although another cytochrome P-450 fraction was active for hexadecane hydroxylation, this fraction had little activity. The results indicate that more than one cytochrome P-450 exists in the intestinal mucosa microsomes.
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D001579:Benzopyrene Hydroxylase / Q000302:isolation & purification / Q000378:metabolism*; D002248:Carbon Monoxide; D002850:Chromatography, Gel; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D006882:Hydroxyapatites; D007413:Intestinal Mucosa / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology*; D009245:NADH Dehydrogenase / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D011817:Rabbits
D006882:Hydroxyapatites; D010718:Phosphatidylserines; D002248:Carbon Monoxide; D003577:Cytochrome P-450 Enzyme System; D001579:Benzopyrene Hydroxylase; D001189:Aryl Hydrocarbon Hydroxylases; D009245:NADH Dehydrogenase