Modulation of water and urea transport in human red cells: effects of pH and phloretin.
It has previously been shown by Macey and Farmer (Biochim. Biophys. Acta 211:104-106, 1970) that phloretin inhibits urea transport across the human red cell membrane yet has no effect on water transport. Jennings and Solomon (J. Gen. Physiol. 67:381-397, 1976) have shown that there are separate lipid and protein binding sites for phloretin on the red cell membrane. We have now found that urea transport is inhibited by phloretin binding to the lipids with a KI of 25 +/- 8 microM in reasonable agreement with the KD of 54 +/- 5 microM for lipid binding. These experiments show that lipid/protein interactions can alter the conformational state of the urea transport protein. Phloretin binding to the protein site also modulates red cell urea transport, but the modulation is opposed by the specific stilbene anion transport inhibitor, DIDS (4,4'-diisothiocyano-2,2'-stilbene disulfonate), suggesting a linkage between the urea transport protein and band 3. Neither the lipid nor the protein phloretin binding site has any significant effect on water transport. Water transport is, however, inhibited by up to 30% in a pH-dependent manner by DIDS binding, which suggests that the DIDS/band 3 complex can modulate water transport.
Department of Physiology and Biophysics, Harvard Medical School, Boston, Massachusetts 02115.
M R
MR
Toon
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Solomon
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D001665:Binding Sites; D001692:Biological Transport / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D003864:Depression, Chemical; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008563:Membrane Lipids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D010693:Phloretin / Q000494:pharmacology*; D014508:Urea / Q000378:metabolism*; D014867:Water / Q000378:metabolism*
D008563:Membrane Lipids; D008565:Membrane Proteins; D014867:Water; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D014508:Urea; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D010693:Phloretin