Properties of an N,N-dimethyl-p-aminoazobenzene oxide reductase purified from rat liver cytosol.
Homogenates of all rat tissues examined, except brain, catalyze reduction of N,N-dimethyl-p-aminoazobenzene N-oxide (DMAB N-oxide) to N,N-dimethyl-p-aminoazobenzene by NADPH. Liver is the most active, and about one third of the homogenate activity of this tissue is recovered in the cytosol fraction. The purified cytosol enzyme has the properties of a tetrameric protein (Mr 370,000) consisting of identical subunits free from chromophores that absorb in the visible spectrum and from metals or other detectable prosthetic groups. The purified reductase is also free from NADPH oxidase and from cytochrome c or azo reductase activities. The enzyme is quite specific for NADPH as reductant and DMAB N-oxide as the electron acceptor. Reduction of other N,N-dimethyl-arylamine or alkylamine oxides as well as N-methylheterocyclicamine oxides could not be detected. Analysis of kinetic data indicate that, at saturating concentrations of the other substrate, 21 microM NADPH and 700 microM DMAB N-oxide are required for half maximal velocity. At infinite concentrations of both substrates the turnover is 150 min-1 at 37 degrees C.
Lashmet Johnson
P R
PR
Clayton Foundation Biochemical Institute, University of Texas, Austin.
Ziegler
D M
DM
D000818:Animals; D003600:Cytosol / Q000201:enzymology; D008099:Liver / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013347:Subcellular Fractions / Q000201:enzymology; D014018:Tissue Distribution; D004124:p-Dimethylaminoazobenzene / Q000493:pharmacokinetics / Q000031:analogs & derivatives
C062917:N,N-dimethyl-4-aminoazobenzene N-oxide; D009249:NADP; D004124:p-Dimethylaminoazobenzene; D009247:NADH, NADPH Oxidoreductases; C019759:azobenzene reductase