Abstract
Four forms of cytochrome P‐450 were separated and purified to electrophoretic homogeneity from human fetal livers. These forms of cytochrome P‐450, termed P‐450HFLa, P‐450HFLb, P‐450HFLc and P‐450HFLd, were distinguishable from each other in their molecular weights, spectral properties, immunochemical properties and mutagen‐producing activities from promntagens. The molecular weights of P‐450HFLa, b, c and d were estimated to be 51,500, 49,000, 51,500 and 50,000, respectively. Antibodies to P‐450HFLa recognized P‐450HFLc but not P‐450HFLb or d, and antibodies to rat P‐448‐H (P‐450IA2) cross‐reacted with P‐450HFLb but not with other forms of cytochrome P‐450. The N‐terminal amino acid sequence of P‐450HFLc was highly homologous, but not identical, to that of P‐450HFLa. Each form of cytochrome P‐450 catalyzed mutagenic activation of aflatoxin Bl (AFB1), 2‐amino‐3‐methylimidazo[4,5‐f]quinoline (IQ) and 2‐amino‐6‐methyldipyrido‐[l,2‐a:3′,2′‐d]imidazole (Glu‐P‐1) at different rates. P‐450 HFLa showed activities to produce mutagen(s) from AFB1, IQ and to a lesser extent from Glu‐P‐1. P‐450 HFLb activated IQ at a faster rate than did the other forms. P‐450 HFLc produced a mutagen from AFB1 and Glu‐P‐1 but not from IQ. P‐450 HFLd did not activate these promutagens at significant rates.
Keywords: Human fetal liver, Cytochrome P‐450, Purification, Promutagens
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In the present study, the apparent molecular weight of P‐450 HFLa was estimated to be 51,500 rather than 51.0006) when bovine serum albumin, catalase and aldolase were used as the standard proteins and when the mobility of P‐450 HFLa was compared to that of P‐450NF.33) We therefore wish to revise the molecular weight of P‐450 HFLa to 51,500.
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