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

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.