Sulfhydryl groups are essential for organic anion exchange in canine renal brush-border membranes.
The effect of several sulfhydryl-modifying reagents (HgCl2, p-chloromercuribenzenesulfonic acid (PCMBS), N-ethylmaleimide) on the renal organic anion exchanger was studied. The transport of p-amino[3H]hippurate, a prototypic organic anion, was examined employing brush-border membrane vesicles isolated from the outer cortex of canine kidneys. HgCl2, PCMBS and N-ethylmaleimide inactivated p-aminohippurate transport with IC50 values of 38, 78 and 190 microM. The rate of p-aminohippurate inactivation by N-ethylmaleimide followed apparent pseudo-first-order reaction kinetics. A replot of the data gave a linear relationship between the apparent rate constants and the N-ethylmaleimide concentration with a slope of 0.8. The data are consistent with a simple bimolecular reaction mechanism and imply that one molecule of N-ethylmaleimide inactivates one essential sulfhydryl group per active transport unit. Substrate (1 mM p-aminohippurate) affected the rate of the N-ethylmaleimide (1.3 mM) inactivation: the t1/2 values for inactivation in the presence and absence of p-aminohippurate were 7.4 and 3.7 min, respectively. The results demonstrate that there are essential sulfhydryl groups for organic anion transport in the brush-border membrane. Moreover, the ability of substrate to alter sulfhydryl reactivity suggests that the latter may play a dynamic role in the transport process.
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000818:Animals; D027321:Anion Transport Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D004285:Dogs; D005033:Ethylmaleimide / Q000494:pharmacology; D007672:Kidney Cortex / Q000378:metabolism*; D007700:Kinetics; D008627:Mercuric Chloride / Q000494:pharmacology; D008871:Microvilli / Q000187:drug effects / Q000378:metabolism*; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D010130:p-Aminohippuric Acid / Q000378:metabolism
D027321:Anion Transport Proteins; D002352:Carrier Proteins; D013439:Sulfhydryl Reagents; D008627:Mercuric Chloride; D002731:4-Chloromercuribenzenesulfonate; D005033:Ethylmaleimide; D010130:p-Aminohippuric Acid