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. 1983 Jan 1;81(1):127–152. doi: 10.1085/jgp.81.1.127

Chloride net efflux from intact erythrocytes under slippage conditions. Evidence for a positive charge on the anion binding/transport site

PMCID: PMC2215566  PMID: 6833995

Abstract

Tracer chloride and potassium net efflux from valinomycin-treated human erythrocytes were measured into media of different chloride concentrations, Clo, at 25 degrees C and pH 7.8. Net efflux was maximal [45-50 mmol (kg cell solids)-1 min-1] at Clo = 0. It decreased hyperbolically with increasing Clo to 14-16 mmol (kg cell solids)-1 min- 1. Half-maximal inhibition occurred at Clo = 3 mM. In the presence of the anion exchange inhibitor DNDS, net efflux was reduced to 5 mmol (kg cell solids)-1 min-1, independent of Clo. Of the three phenomenological components of net efflux, the Clo-inhibitable (DNDS-inhibitable) component was tentatively attributed to "slippage," that is, net transport mediated by the occasional return of the empty transporter. The Clo-independent (DNDS-inhibitable) component was tentatively attributed to movement of chloride through the anion transporter without the usual conformational change of the transport site on the protein ("tunneling"). These concepts of slippage and tunneling are shown to be compatible with a model that describes the anion transporter as a specialized single-site, two-barrier channel that can undergo conformational changes between two states. Net chloride efflux when the slippage component dominated (Clo = 0.7 mM) was accelerated by a more negative (inside) membrane potential. It appears that the single anion binding-and-transport site on each transporter has one net positive charge and that is neutralized when a chloride ion is bound.

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Selected References

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  1. Barzilay M., Ship S., Cabantchik Z. I. Anion transport in red blood cells. I. Chemical properties of anion recognition sites as revealed by structure-activity relationships of aromatic sulfonic acids. Membr Biochem. 1979;2(2):227–254. doi: 10.3109/09687687909063866. [DOI] [PubMed] [Google Scholar]
  2. Brahm J. Temperature-dependent changes of chloride transport kinetics in human red cells. J Gen Physiol. 1977 Sep;70(3):283–306. doi: 10.1085/jgp.70.3.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cass A., Dalmark M. Chloride transport by self-exchange and by KCl salt diffusion in gramicidin-treated red blood cells. Acta Physiol Scand. 1979 Nov;107(3):193–203. doi: 10.1111/j.1748-1716.1979.tb06463.x. [DOI] [PubMed] [Google Scholar]
  4. Dalmark M. Chloride transport in human red cells. J Physiol. 1975 Aug;250(1):39–64. doi: 10.1113/jphysiol.1975.sp011042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dalmark M. Effects of halides and bicarbonate on chloride transport in human red blood cells. J Gen Physiol. 1976 Feb;67(2):223–234. doi: 10.1085/jgp.67.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dalmark M., Wieth J. O. Temperature dependence of chloride, bromide, iodide, thiocyanate and salicylate transport in human red cells. J Physiol. 1972 Aug;224(3):583–610. doi: 10.1113/jphysiol.1972.sp009914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fröhlich O. The external anion binding site of the human erythrocyte anion transporter: DNDS binding and competition with chloride. J Membr Biol. 1982;65(1-2):111–123. doi: 10.1007/BF01870474. [DOI] [PubMed] [Google Scholar]
  8. Geck P. Eigenschaften eines asymmetrischen Carrier-Modells für den Zuckertrnasport am menschlichen Erythrozyten. Biochim Biophys Acta. 1971 Aug 13;241(2):462–472. doi: 10.1016/0005-2736(71)90045-9. [DOI] [PubMed] [Google Scholar]
  9. Grinstein S., McCulloch L., Rothstein A. Transmembrane effects of irreversible inhibitors of anion transport in red blood cells. Evidence for mobile transport sites. J Gen Physiol. 1979 Apr;73(4):493–514. doi: 10.1085/jgp.73.4.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gunn R. B., Dalmark M., Tosteson D. C., Wieth J. O. Characteristics of chloride transport in human red blood cells. J Gen Physiol. 1973 Feb;61(2):185–206. doi: 10.1085/jgp.61.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gunn R. B., Fröhlich O. Asymmetry in the mechanism for anion exchange in human red blood cell membranes. Evidence for reciprocating sites that react with one transported anion at a time. J Gen Physiol. 1979 Sep;74(3):351–374. doi: 10.1085/jgp.74.3.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hladky S. B., Haydon D. A. Ion transfer across lipid membranes in the presence of gramicidin A. I. Studies of the unit conductance channel. Biochim Biophys Acta. 1972 Aug 9;274(2):294–312. doi: 10.1016/0005-2736(72)90178-2. [DOI] [PubMed] [Google Scholar]
  13. Hoffman J. F., Laris P. C. Determination of membrane potentials in human and Amphiuma red blood cells by means of fluorescent probe. J Physiol. 1974 Jun;239(3):519–552. doi: 10.1113/jphysiol.1974.sp010581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hunter M. J. A quantitative estimate of the non-exchange-restricted chloride permeability of the human red cell. J Physiol. 1971 Oct;218 (Suppl):49P–50P. [PubMed] [Google Scholar]
  15. Hunter M. J. Human erythrocyte anion permeabilities measured under conditions of net charge transfer. J Physiol. 1977 Jun;268(1):35–49. doi: 10.1113/jphysiol.1977.sp011845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jennings M. L. Stoichiometry of a half-turnover of band 3, the chloride transport protein of human erythrocytes. J Gen Physiol. 1982 Feb;79(2):169–185. doi: 10.1085/jgp.79.2.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kaplan J. H., Passow H. Effects of phlorizin on net chloride movements across the valinomycin-treated erythrocyte membrane. J Membr Biol. 1974;19(1):179–194. doi: 10.1007/BF01869977. [DOI] [PubMed] [Google Scholar]
  18. Knauf P. A., Fuhrmann G. F., Rothstein S., Rothstein A. The relationship between anion exchange and net anion flow across the human red blood cell membrane. J Gen Physiol. 1977 Mar;69(3):363–386. doi: 10.1085/jgp.69.3.363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Knauf P. A., Law F. Y., Marchant P. J. Relationship of net chloride flow across the human erythrocyte membrane to the anion exchange mechanism. J Gen Physiol. 1983 Jan;81(1):95–126. doi: 10.1085/jgp.81.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lambert A., Lowe A. G. Chloride/bicarbonate exchange in human erythrocytes. J Physiol. 1978 Feb;275:51–63. doi: 10.1113/jphysiol.1978.sp012177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lieb W. R., Stein W. D. Testing and characterizing the simple carrier. Biochim Biophys Acta. 1974 Dec 10;373(2):178–196. doi: 10.1016/0005-2736(74)90144-8. [DOI] [PubMed] [Google Scholar]
  22. Läuger P. Kinetic properties of ion carriers and channels. J Membr Biol. 1980 Dec 30;57(3):163–78(-RETURN-). doi: 10.1007/BF01869585. [DOI] [PubMed] [Google Scholar]
  23. REGEN D. M., MORGAN H. E. STUDIES OF THE GLUCOSE-TRANSPORT SYSTEM IN THE RABBIT ERYTHROCYTE. Biochim Biophys Acta. 1964 Jan 27;79:151–166. doi: 10.1016/0926-6577(64)90048-8. [DOI] [PubMed] [Google Scholar]
  24. Rothstein A., Cabantchik Z. I., Knauf P. Mechanism of anion transport in red blood cells: role of membrane proteins. Fed Proc. 1976 Jan;35(1):3–10. [PubMed] [Google Scholar]
  25. WILKINSON G. N. Statistical estimations in enzyme kinetics. Biochem J. 1961 Aug;80:324–332. doi: 10.1042/bj0800324. [DOI] [PMC free article] [PubMed] [Google Scholar]

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