Control of red cell urea and water permeability by sulfhydryl reagents.
The binding constant for pCMBS (p-chloromercuribenzenesulfonate) inhibition of human red cell water transport has been determined to be 160 +/- 30 microM and that for urea transport inhibition to be 0.09 +/- 0.06 microM, indicating that there are separate sites for the two inhibition processes. The reaction kinetics show that both processes consist of a bimolecular association between pCMBS and the membrane site followed by a conformational change. Both processes are very slow and the on rate constant for the water inhibition process is about 10(5) times slower than usual for inhibitor binding to membrane transport proteins. pCMBS binding to the water transport inhibition site can be reversed by cysteine while that to the urea transport inhibition site can not be reversed. The specific stilbene anion exchange inhibitor, DBDS (4,4'-dibenzamidostilbene-2,2'-disulfonate) causes a significant change in the time-course of pCMBS inhibition of water transport, consistent with a linkage between anion exchange and water transport. Consideration of available sulfhydryl groups on band 3 suggests that the urea transport inhibition site is on band 3, but is not a sulfhydryl group, and that, if the water transport inhibition site is a sulfhydryl group, it is located on another protein complexed to band 3, possibly band 4.5.
D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D002731:4-Chloromercuribenzenesulfonate / Q000378:metabolism / Q000494:pharmacology; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D001834:Body Water / Q000378:metabolism*; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003545:Cysteine / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D011487:Protein Conformation / Q000187:drug effects; D013267:Stilbenes / Q000037:antagonists & inhibitors; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D014508:Urea / Q000097:blood*
D001457:Anion Exchange Protein 1, Erythrocyte; D013267:Stilbenes; D013439:Sulfhydryl Reagents; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D002731:4-Chloromercuribenzenesulfonate; C036010:4,4'-dibenzamido-2,2'-stilbenedisulfonic acid; D014508:Urea; D003545:Cysteine