Inhibition of Anion Permeability by Amphiphilic Compounds in Human Red Cell: Evidence for an Interaction of Niflumic Acid with the Band 3 Protein

Overview

Journal J Membr Biol

Date 1979 Apr 20

PMID 376851

Citations 23

Authors

J L Cousin

R MOTAIS

Affiliations

Soon will be listed here.

Abstract

In human erythrocyte, permeability to the anion is instantaneously, reversibly, and noncompetitively inhibited by the nonsteroidal anti-inflammatory drug, niflumic acid. The active form of this powerful inhibitor (I50 = 6 X 10(-7) M) is the ionic form. We demonstrated that: (i) The binding of niflumic acid to the membrane of unsealed ghosts show one saturable and one linear component over the concentration range studied. The saturable component vanishes when chloride transport is fully inhibited by covalently bound 4-acetamido-4'-isothiocyano stilbene-2,2'-disulfonic acid (SITS). Our estimate of these SITS protectable niflumate binding sites (about 9 x 10(5) per cell) agrees with the number of protein molecules per cell in band 3. These sites are half-saturated with 10(-6) M niflumic acid, a concentration very close to I50. (ii) Niflumic acid inhibits the binding reaction of SITS with anion controlling transport sites. These results indicate that niflumic acid and SITS are mutually exclusive inhibitors, suggesting that niflumic acid interacts with the protein in band 3. Niflumic acid also decreases glucose and ouabain-insensitive sodium permeabilities. However, these effects are produced at a very high concentration of niflumic acid (in millimolar range), suggesting unspecific action, possibly through lipid phase.

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References

Rothstein A, Cabantchik Z, Knauf P . Mechanism of anion transport in red blood cells: role of membrane proteins. Fed Proc. 1976; 35(1):3-10. View

McLaughlin S . Salicylates and phospholipid bilayer membranes. Nature. 1973; 243(5404):234-6. DOI: 10.1038/243234a0. View

Lefevre P . Sugar transport in the red blood cell: structure-activity relationships in substrates and antagonists. Pharmacol Rev. 1961; 13:39-70. View

Andersen O, Finkelstein A, Katz I, Cass A . Effect of phloretin on the permeability of thin lipid membranes. J Gen Physiol. 1976; 67(6):749-71. PMC: 2214976. DOI: 10.1085/jgp.67.6.749. View

Fairbanks G, Steck T, Wallach D . Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971; 10(13):2606-17. DOI: 10.1021/bi00789a030. View

Dunn M . The effects of transport inhibitors on sodium outflux and influx in red blood cells: evidence for exchange diffusion. J Clin Invest. 1970; 49(10):1804-14. PMC: 322669. DOI: 10.1172/JCI106398. View

Zaki L, FASOLD H, Schuhmann B, Passow H . Chemical modification of membrane proteins in relation to inhibition of anion exchange in human red blood cells. J Cell Physiol. 1975; 86(3 Pt 1):471-94. DOI: 10.1002/jcp.1040860305. View

Passow H, FASOLD H, Lepke S, Pring M, Schuhmann B . Chemical and enzymatic modification of membrane proteins and anion transport in human red blood cells. Adv Exp Med Biol. 1977; 84:353-79. DOI: 10.1007/978-1-4684-3279-4_17. View

MOTAIS R, Cousin J . [Demonstration of the inhibitory effect of ethacrynic acid on permeability to chlorine]. C R Acad Hebd Seances Acad Sci D. 1976; 283(1):63-5. View

10.

Lin S, Spudich J . Binding of cytochalasin B to a red cell membrane protein. Biochem Biophys Res Commun. 1974; 61(4):1471-6. DOI: 10.1016/s0006-291x(74)80449-3. View

11.

Schnell K . On the mechanism of inhibition of the sulfate transfer across the human erythrocyte membrane. Biochim Biophys Acta. 1972; 282(1):265-76. DOI: 10.1016/0005-2736(72)90333-1. View

12.

Cass A, Dalmark M . Equilibrium dialysis of ions in nystatin-treated red cells. Nat New Biol. 1973; 244(132):47-9. DOI: 10.1038/newbio244047a0. View

13.

DODGE J, Mitchell C, HANAHAN D . The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963; 100:119-30. DOI: 10.1016/0003-9861(63)90042-0. View

14.

Cabantchik Z, Balshin M, Breuer W, Markus H, Rothstein A . A comparison of intact human red blood cells and resealed and leaky ghosts with respect to their interactions with surface labelling agents and proteolytic enzymes. Biochim Biophys Acta. 1975; 382(4):621-33. DOI: 10.1016/0005-2736(75)90227-8. View

15.

Passow H, Schnell K . Chemical modifiers of passive ion permeability of the erythrocyte membrane. Experientia. 1969; 25(5):460-8. DOI: 10.1007/BF01900757. View

16.

Brown P, Feinstein M, Shaafi R . Membrane proteins related to water transport in human erythrocytes. Nature. 1975; 254(5500):523-5. DOI: 10.1038/254523a0. View

17.

Cabantchik Z, Rothstein A . The nature of the membrane sites controlling anion permeability of human red blood cells as determined by studies with disulfonic stilbene derivatives. J Membr Biol. 1972; 10(3):311-30. DOI: 10.1007/BF01867863. View

18.

Sheetz M, Singer S . Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions. Proc Natl Acad Sci U S A. 1974; 71(11):4457-61. PMC: 433905. DOI: 10.1073/pnas.71.11.4457. View

19.

Rice W, Steck T . Pyruvate flux into resealed ghosts from human erythrocytes. Biochim Biophys Acta. 1976; 433(1):39-53. DOI: 10.1016/0005-2736(76)90176-0. View

20.

DEUTICKE B, Rickert I, Beyer E . Stereoselective, SH-dependent transfer of lactate in mammalian erythrocytes. Biochim Biophys Acta. 1978; 507(1):137-55. DOI: 10.1016/0005-2736(78)90381-4. View