In vitro methylation of inorganic arsenic in mouse intestinal cecum.
The capacity of mouse intestinal cecal microflora to methylate inorganic arsenicals (iAs) was examined in vitro under conditions of restricted bacterial growth. Cecal contents incubated under anaerobic conditions at 37 degrees C for 21 hr methylated up to 40% of either 0.1 microM arsenite (iAsIII) or 0.1 microM arsenate (iAsV). Methylarsenic (MAs) was the predominant metabolite; however, about 3% of either substrate was converted to dimethylarsenic (DMAs). Over the first 6 hr, the rate of methylation was several times greater for iAsIII than for iAsV. There was a 3-hr delay in the production of methylated metabolites from iAsV, suggesting that reduction of iAsV to iAsIII before methylation could be rate limiting. Over the concentration range of 0.1 to 10 microM of iAsIII or iAsV, there was an approximately linear increase in the production of MAs and DMAs. There was evidence of saturation or inhibition of methylation at 100 microM of either substrate. Substrate concentration had little effect on MAs/DMAs ratio. Incubation of cecal contents at 0 degrees C abolished methylation of either arsenical. Under aerobic or anaerobic conditions, cecal tissue homogenates produced little MAs or DMAs from either arsenical. Addition of potential methyl group donors, L-methionine and methylcobalamin, into cecal contents significantly increased the rate of methylation, especially for iAsV. Addition of glutathione, but not L-cysteine, had a similar effect. Selenite, a recognized inhibitor of iAs methylation in mammalian tissues, inhibited methylation of either substrate by cecal contents. These data suggest that cecal microflora are a high capacity methylation system that might contribute significantly to methylation of iAs in intact animals.
Hall
L L
LL
Experimental Toxicology Division, United States Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA.
George
S E
SE
Kohan
M J
MJ
Styblo
M
M
Thomas
D J
DJ
D000818:Animals; D020261:Arsenic Poisoning; D001152:Arsenicals / Q000737:chemistry / Q000378:metabolism*; D002432:Cecum / Q000378:metabolism / Q000382:microbiology*; D002855:Chromatography, Thin Layer; D005978:Glutathione / Q000737:chemistry; D006090:Gram-Negative Bacteria / Q000187:drug effects*; D006094:Gram-Positive Bacteria / Q000187:drug effects*; D008297:Male; D008715:Methionine / Q000737:chemistry; D008745:Methylation / Q000187:drug effects; D051379:Mice; D018038:Sodium Selenite / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000494:pharmacology; D014805:Vitamin B 12 / Q000031:analogs & derivatives / Q000737:chemistry
D001152:Arsenicals; D013438:Sulfhydryl Compounds; D008715:Methionine; C019476:mecobalamin; D005978:Glutathione; D018038:Sodium Selenite; D014805:Vitamin B 12