New evidence for the essential role of arginine residues in anion transport across the red blood cell membrane.
2,3-Butanedione, in the dark and in the presence of borate, reacts rapidly to inactivate the sulfate equilibrium exchange across the human red cell membrane. Reactivation occurs spontaneously after the removal of borate, indicating the reaction of butanedione with essential arginine residues. The inactivation of the transport system depends on the concentration of the reagent, on the incubation time and exhibits pseudo-first-order kinetics. Chloride ions are able to protect the transport system against inactivation with the reagent. This would suggest the participation of the modified residue in the substrate binding site. When the transport system is inhibited to 50-60% by butanedione, the transporter can still bind covalently the anion transport inhibitor 2H2DIDS up to 85 +/- 12% of its total binding capacity. 3H2DIDS concentration was either 3.15, 10 or 20 microM. Modification of resealed ghosts with 50 mM butanedione under conditions where the transport system is to more than 75% inhibited, causes a reduction of only about 30% of the reversibly bound 3H2DIDS.
D027321:Anion Transport Proteins; D001120:Arginine; D002074:Butanones / Q000494:pharmacology*; D002352:Carrier Proteins / Q000097:blood*; D002712:Chlorides / Q000494:pharmacology; D003931:Diacetyl / Q000494:pharmacology*; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D013431:Sulfates / Q000097:blood
D027321:Anion Transport Proteins; D002074:Butanones; D002352:Carrier Proteins; D002712:Chlorides; D013431:Sulfates; D001120:Arginine; D003931:Diacetyl