Springer Nature is making Coronavirus research free. View research | View latest news | Sign up for updates

Action of 1-fluoro-2,4-dinitrobenzene on passive ion permeability of the human red blood cell

  • 28 Accesses

  • 46 Citations

Summary

Dinitrofluorobenzene (DNFB) inhibits the penetration of anions such as sulfate, phosphate, succinate, and lactate, and facilitates the penetration of cations such as K+ and Na+. The phlorizin-glucose insensitive fraction of erythritol permeability is not affected by the agent. The effects of DNFB on ion permeability are similar to those of more specific amino reactive agents like trinitrobenzene sulfonate and 2-methoxy-5-nitrotropone.

Anion permeability reacts more sensitively to DNFB than cation permeability. At a given concentration of DNFB in the medium, the inhibition of anion permeability develops faster than the facilitation of cation permeability. At a given time of exposure, lower concentrations of DNFB are required to produce a nearly maximal response of anion permeability than are necessary for maximal effect on cation permeability.

The response of anion and cation permeability to DNFB is augmented by increasing the pH at which dinitrophenylation is allowed to take place.

DNFB binding to the cell membrane is about one order of magnitude lower than DNFB binding to the whole cell. In the cell membrane, proteins as well as lipids are dinitrophenylated. Among the lipids, only phosphatidylethanolamine binds significant amounts of DNFB. Phosphatidylserine does not seem to react with the agent under the experimental conditions under which DNFB produces its effects on ion permeability.

The experimental results are compatible with the assumption that removal of uncharged NH2-groups by dinitrophenylation of the membrane leads to a concomitant reduction of fixed NH 3 + -groups and hence of the positive membrane charge. This leads to an acceleration of cation movements and an inhibition of anion permeability while nonelectrolyte permeability remains unaffected. However, other explanations of our observations cannot be ruled out.

This is a preview of subscription content, log in to check access.

References

  1. Bartlett, G. R. 1959. Phosphorus assay in column chromatography.J. Biol. Chem. 234:466.

  2. Berg, H. C., Diamond, J. M., Marfey, P. S. 1965. Erythrocyte membrane: chemical modification.Science 150:64.

  3. Bowyer, F., Widdas, W. F. 1955. Erythrocyte permeability to erythritol.J. Physiol. 129:7P.

  4. ——, 1956. The facilitated transfer of glucose and related compounds across the erythrocyte membrane.Disc. Faraday Soc. 21:251.

  5. Dodge, J. T., Mitchell, C., Hanahan, D. 1962. The preparation and chemical characteristics of hemoglobin free ghosts of human erythrocytes.Arch. Biochem. Biophys. 100:119.

  6. Gardos, G., Hoffman, J. F., Passow, H. 1969. Flux measurements in erythrocytes.In: Laboratory Techniques in Membrane Biophysics. H. Passow and R. Stämpfli, editors. p. 9. Springer-Verlag, Berlin-Heidelberg-New York.

  7. Henkart, P., Guidotti, G., Edsall, J. T. 1968. Catalysis of the hydrolysis of 1-fluoro-2,4-dinitrobenzene by carbonic anhydrase.J. Biol. Chem. 243:2447.

  8. Hirs, C. H. W. 1967. Reactions with reactive aryl halides.In: Methods in Enzymology, vol. XI, p. 548. C. H. W. Hirs, editor. Academic Press, New York.

  9. Knauf, P., Rothstein, A. 1968.Quoted in: A. Rothstein. Membrane permeability of erythrocytes.In: Metabolism and Membrane Permeability of Erythrocytes. E. Deutsch, E. Gerlach, and K. Moser, editors. p. 407. Thieme Verlag, Stuttgart.

  10. LaCelle, P., Passow, H. 1971. Permeability of the human red blood cell tomeso-erythritol.J. Membrane Biol. 4:270.

  11. Lepke, S., Passow, H. 1971. The permeability of the human red blood cell to sulfate ions.J. Membrane Biol. 6:158.

  12. Parpart, A. K., Ballantine, R. 1952. Molecular anatomy of the red cell plasma membrane.In: Modern Trends in Physiology and Biochemistry. E. S. G. Barran, editor. p. 135. Academic Press, New York.

  13. Passow, H. 1969a. Passive permeability of the erythrocyte membrane. — An assessment of scope and limitations of the fixed charge hypothesis.Progr. Biophys. Mol. Biol. 19:425.

  14. — 1969b. Ion permeability of erythrocyte ghosts.In: Laboratory Techniques in Membrane Biophysics. H. Passow and R. Stämpfli, editors. p. 21. Springer-Verlag, Berlin-Heidelberg-New York.

  15. — 1971. Effects of pronase on the passive ion permeability of the human red blood cell.J. Membrane Biol. 6:233.

  16. —, Schnell, K. F. 1969. Chemical modifiers of passive ion permeability of the erythrocyte membrane.Experientia 25:460.

  17. Reed, S. F., Scott, N., Swisher, S. N., Marinetti, K. V., Eden, E. K. 1960. Studies of the lipids of the erythrocyte. I. Quantitative analysis of the lipids of normal human red blood cells.J. Lab. Clin. Med. 56:281

  18. Van Deenen, L. L. M. 1969. Membrane lipids and lipophilic proteins.In: The Molecular Basis of Membrane Function. D. C. Tosteson, editor. p. 47. Prentice-Hall, Inc., Englewood Cliffs, N.T.

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Poensgen, J., Passow, H. Action of 1-fluoro-2,4-dinitrobenzene on passive ion permeability of the human red blood cell. J. Membrain Biol. 6, 210–232 (1971). https://doi.org/10.1007/BF01872278

Download citation

Keywords

  • Succinate
  • Phosphatidylethanolamine
  • Phosphatidylserine
  • Erythritol
  • Concomitant Reduction