ORGANIC-ANIONS EXHIBIT DISTINCT INHIBITION PATTERNS ON THE LOW-KM AND HIGH-KM TRANSPORT OF S-(2,4-DINITROPHENYL)GLUTATHIONE THROUGH THE HUMAN ERYTHROCYTE-MEMBRANE

The low-K(m) and high-K(m) components of S-dinitrophenyl-glutathione (DNPSG) uptake by inside-out vesicles of human erythrocytes show different pH profiles and inhibition properties with organic anions. Both components are competitively inhibited by various polyvalent anions, including glutathione conjugates, conjugated steroid hormones and bile salts, and bilirubin ditaurate. A variety of monovalent anions, including glucuronidated and sulphated drugs and taurocholate, inhibit the high-K(m) system only. Taurocholate is taken up by the erythrocyte vesicles in an ATP-dependent manner. The anionic dyes fluorescein, Indocyanine Green and bromosulphophthalein inhibit the low-K(m) system competitively and the high-K(m) system non-competitively. The study shows that interactions between different types of biologically occurring conjugates can occur at the level of the transport step out of erythrocytes. The kinetic properties suggest overlapping substrate specificities for the two systems, in which the low-K(m) component is physiologically more important for transport of glutathione conjugates and polyvalent organic anions, whereas the high-K(m) component is of significance for transport of monovalent organic anions. Low- and high-K(m) transport of DNPSG is also observed in plasma membrane vesicles from rat, pig and bovine erythrocytes.