The Journal of Membrane Biology

, Volume 102, Issue 3, pp 217–224 | Cite as

Studies on inactivation of anion transport in human red blood cell membrane by reversibly and irreversibly acting arginine-specific reagents

  • T. Julien
  • L. Zaki


A chromophoric derivative of phenylglyoxal, 4-hydroxy-3-nitrophenylglyoxal (HNPG), known to be highly selective for modification of arginine residues in aqueous solution is found to be a potent inhibitor of anion transport across the red cell membrane. In contrast to the action of all other arginine-specific reagents used under the experimental conditions in this laboratory, the action of HNPG on sulfate transport is completely reversible. Hence, a kinetic analysis of its inhibitory effect on SO 4 2− self-exchange could be performed. The effect of increasing chloride concentration on the inhibitory potency of HNPG is consistent with the concept that Cl and HNPG compete for the same site on the anion transporter. The IC50 value for the inhibition of SO 4 2− exchange with HNPG is about 0.13mm at pH 8.0 and 0.36mm at pH 7.4, and the Hill coefficient for the interaction between the transporter and the inhibitor is near one at both pH's. HNPG is able to protect the transport system against inhibition with the (under our experimental conditions) irreversibly acting arginine specific reagent, phenylglyoxal. Partial inactivation of the transport system with phenylglyoxal lowers the maximal rates of SO 4 2− and chloride exchange but does not modify the apparentK s for the substrate anions. Reversibly acting anion transport inhibitors known to interact with the DIDS binding site like salicylate, tetrathionate, APMB, DNDS, and flufenamate are able to protect the transport system against phenylglyoxalation. Other inhibitors like phloretin and phlorizin have no effect.

Key Words

erythrocyte anion transporter arginine substrate binding site 


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Copyright information

© Springer-Verlag New York Inc 1988

Authors and Affiliations

  • T. Julien
    • 1
  • L. Zaki
    • 1
  1. 1.Max-Planck-Institut für BiophysikFrankfurtWest Germany

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