Journal
CHEMICAL RESEARCH IN TOXICOLOGY
Volume 14, Issue 6, Pages 727-737Publisher
AMER CHEMICAL SOC
DOI: 10.1021/tx010005p
Keywords
-
Funding
- NCI NIH HHS [R01 CA39416, R35 CA44353] Funding Source: Medline
- NIEHS NIH HHS [F32 ES05919, P30 ES00267] Funding Source: Medline
Ask authors/readers for more resources
Oxidation of the mycotoxin aflatoxin (AF) B-1 yields the 8,9-epoxide, which nonenzymatically hydrolyzes rapidly to a dihydrodiol that in turn undergoes slow, base-catalyzed ring opening to a dialdehyde [Johnson, W. W., Harris, T. M., and Guengerich F. P. (1996) J. Ant. Chem. Sec. 118, 8213-8220]. AFB(1) dialdehyde does not bind to DNA but can react with protein lysine groups. One enzyme induced by cancer chemopreventive agents is AFB(1) aldehyde reductase (AFAR), which catalyzes the NADPH-dependent reduction of the dialdehyde to a dialcohol. AFB(1) dialdehyde is known to convert, nonenzymatically to AFB(1) dihydrodiol at neutral pH, and we reinvestigated the enzymatic reaction by preparing AFB(1) dialdehyde at pH 10 and then used this to initiate reactions (at neutral pH) with rat and human AFAR isozymes. Two monoalcohols were identified as products, and their identities were established by (NaBH4)-H-2 reduction, chemical cleavage, and mass spectrometry. The monoalcohol corresponding to reduction at C-8 formed first in reactions catalyzed by either the rat or the human AFAR. This C-8 monoalcohol was further reduced to AFB(1) dialcohol by AFAR. The other monoalcohol (C-6a) was formed but not reduced to the dialcohol rapidly. Steady-state kinetic parameters were Estimated for the reduction of AFB(1) dialdehyde by rat and human AFAR to the monoalcohols. The apparent k(cat) and K-m values were not adequate to rationalize the observed DeltaA(340) spectral changes in a kinetic model. Simulation fitting was done and yielded parameters indicative of greater enzyme efficiency. A survey of 12 human liver cytosol samples showed a variation of 2.3-fold in AFAR activity. Rats treated with AFB(1) excreted the dialcohol and a monoalcohol in urine. The results of these studies are consistent with a role of(rat and human) AFAR in protection against AFB(1) toxicity.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available