Steroidal nonspecific esterase metabolism of N-hydroxy-2-acetylaminofluorene: evidence for selective activation by the cellular reductant NADPH.
The significance of the nonspecific esterases of human mononuclear leukocytes (HMLs) in arylamine carcinogenesis is suggested by data showing that the metabolically formed hydroxamic acid derivative of 2-acetylaminofluorene, N-hydroxy-2-acetylaminofluorene, is a substrate for this class of enzymes. A viable cell assay for the nonspecific esterases using alpha-naphthyl acetate as substrate is described, and data showing this activity to be sensitive to already known substrates for HML esterases as measured by three previously described assays are presented. All four assays of the same esterase activity are shown to be highly sensitive to up- and down-regulation by addition of NADPH or NADP to viable HML cultures. Selective activation of a purified rabbit nonspecific esterase by NADPH, but not by the other cellular reductants, NADH and glutathione, was demonstrated. Cytosols prepared from normal human tissue samples of liver, breast, colon, and brain were also activated by the presence of NADPH. These data do not indicate that steroidal nonspecific esterases are redox-modulated by the presence of mixed disulfides in their structure. Instead, they support the direct and specific influence of NADPH as a widespread activator of esterase activity by a mechanism not yet understood.
Lund-Pero
M
M
Preventive Medicine Institute, Strang Clinic, Division of Biochemical Epidemiology, New York, New York.
Pero
R W
RW
Miller
D G
DG
D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000737:chemistry*; D004249:DNA Damage; D004789:Enzyme Activation; D004950:Esterases / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006881:Hydroxyacetylaminofluorene / Q000737:chemistry / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism*; D010084:Oxidation-Reduction; D011817:Rabbits; D013256:Steroids / Q000378:metabolism
D013256:Steroids; D009243:NAD; D009249:NADP; D006881:Hydroxyacetylaminofluorene; D004247:DNA; D004950:Esterases; D005978:Glutathione