Modification of a carboxyl group that appears to cross the permeability barrier in the red blood cell anion transporter.
A recently developed method for converting protein carboxyl groups to alcohols has been used to examine the functional role of carboxyl groups in the red blood cell inorganic anion-transport protein (band 3). A major goal of the work was to investigate the carboxyl group that is protonated during the proton-sulfate cotransport that takes place during net chloride-sulfate exchange. Three kinds of evidence indicate that the chemical modification (Woodward's reagent K followed by borohydride) converts this carboxyl to an alcohol. First, monovalent anion exchange is inhibited irreversibly. Second, the modification stimulates sulfate influx into chloride-loaded cells and nearly eliminates the extracellular pH dependence of the sulfate influx. (The stimulated sulfate influx in the modified cells is inhibitable by stilbenedisulfonate.) Third, the proton influx normally associated with chloride-sulfate exchange is inhibited by the modification. These results would all be expected if the titratable carboxyl group were converted into the untitratable, neutral alcohol. In addition to altering the extracellular pH dependence of sulfate influx, the chemical modification removes the intracellular pH dependence of sulfate efflux. The modification is performed under conditions in which the reagent does not cross the permeability barrier. The large effect on the intracellular pH dependence of sulfate transport suggests that a single carboxyl group can at different times be in contact with the aqueous medium on each side of the permeability barrier.
Jennings
M L
ML
Department of Physiology and Biophysics, University of Texas Medical Branch, Galveston 77550.
Al-Rhaiyel
S
S
D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D000838:Anions; D002264:Carboxylic Acids; D002463:Cell Membrane Permeability; D002712:Chlorides / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007474:Ion Exchange; D013431:Sulfates / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D000838:Anions; D002264:Carboxylic Acids; D002712:Chlorides; D013431:Sulfates