The interaction of phenyldichloroarsine with erythrocytes.
The purpose of the study was to identify binding sites of organic arsenic in the erythrocyte and to explain species differences in binding. Washed erythrocytes were exposed to graded concentrations of [U-14C]phenyldichloroarsine (PDA) in phosphate-buffered saline containing 0.1% glucose and 0.1% bovine serum albumin. At low PDA concentrations, all cells bound the arsenical rapidly (within 10 min) and quantitatively. Human, pig, hamster, guinea pig, and mouse erythrocytes approached saturation at 0.02-0.3 mumol PDA/10(9) cells, depending on the species. Saturation points correlated well with each respective species' erythrocyte glutathione content. In contrast, rat erythrocytes showed no sign of saturation at PDA loads as high as 3.0 mumol/10(9) cells. Hemolysates of PDA-treated erythrocytes were subjected to Sephadex G-75 gel filtration chromatography. 14C from rat hemolysate was distributed between the hemoglobin and small molecular weight (glutathione-containing) fractions. In all other species, the 14C eluted almost exclusively with the glutathione-containing fractions. In equilibrium dialysis experiments, human hemoglobin did not bind PDA, whereas rat hemoglobin bound 2 PDA/mol with Kd approximately 5 microM. In conclusion, glutathione is the principal binding site of phenyldichloroarsine in erythrocytes. In most species, the arsenical does not bind to hemoglobin, even though it has free (titratable) sulfhydryls considerably in excess of the glutathione concentration. In rat erythrocytes, phenlydichloroarsine binds both to glutathione and to hemoglobin. Arsenical binding by rat hemoglobin is presumably due to the unique location of the extra titratable cysteine in that protein.
Division of Biophysical Research, Letterman Army Institute of Research, Presidio of San Francisco, California 94129-6800.
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Chong
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Dill
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McGown
D000818:Animals; D020261:Arsenic Poisoning; D001152:Arsenicals / Q000097:blood*; D002850:Chromatography, Gel; D006224:Cricetinae; D003956:Dialysis; D004912:Erythrocytes / Q000378:metabolism*; D006168:Guinea Pigs; D006454:Hemoglobins / Q000378:metabolism; D008297:Male; D008647:Mesocricetus; D051379:Mice; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains
D001152:Arsenicals; D006454:Hemoglobins; C053663:phenyldichloroarsine