Adhesion of human red blood cells to polystyrene. Influence of sodium chloride concentration and of neuraminidase treatment

Chloride self-exchange was determined by measuring the rate of 36Cl efflux from human red blood cells at pH 7.2 (0 degrees C) in the presence of fluoride, bromide, iodide, and bicarbonate. The chloride concentration was varied between 10--400 mM and the concentration of other halides and bicarbonate between 10--300 mM. Chloride equilibrium flux showed saturation kinetics. The half-saturation constant increased and the maximum flux decreased in the presence of halides and bicarbonate: the inhibition kinetics were both competitive and noncompetitive. The competitive and the noncompetitive effects increased proportionately in the sequence: fluoride less than bromide less than iodide. The inhibitory action of bicarbonate was predominantly competitive. The noncompetitive effect of chloride (chloride self-inhibition) on chloride transport was less dominant at high inhibitor concentrations. Similarly, the noncompetitive action of the inhibitors was less dominant at high chloride concentrations. The results can be described by a carrier model with two anion binding sites: a transport site, and a second site which modifies the maximum transport rate. Binding to both types of sites increases proportionately in the sequence: fluoride less than chloride less than bromide less than iodide.

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Kinetics of residual chloride transport in human red blood cells after maximum covalent 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid binding.

The Journal of General Physiology ◽

10.1085/jgp.101.5.715 ◽

1993 ◽

Vol 101 (5) ◽

pp. 715-732 ◽

Cited By ~ 5

Author(s):

P K Gasbjerg ◽

J Funder ◽

J Brahm

Keyword(s):

Red Blood Cells ◽

Lipid Bilayers ◽

Blood Cells ◽

Chloride Transport ◽

Covalent Binding ◽

Chloride Concentration ◽

Disulfonic Acid ◽

Human Red Blood Cells ◽

O 19 ◽

Kinetics Of

Irreversible inhibition, 99.8% of control values for chloride transport in human red blood cells, was obtained by well-established methods of maximum covalent binding of 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). The kinetics of the residual chloride transport (0.2%, 106 pmol.cm-2 x s-1) at 38 degrees C, pH 7.2) was studied by means of 36Cl- efflux. The outside apparent affinity, expressed by Ko1/2,c, was 34 mM, as determined by substituting external KCl by sucrose. The residual flux was reversibly inhibited by a reexposure to DIDS, and by 4,4'-dinitrostilbene-2,2'-disulfonate (DNDS), phloretin, salicylate, and alpha-bromo-4-hydroxy-3,5-dinitroacetophenone (Killer III) (Borders, C. L., Jr., D. M. Perez, M. W. Lafferty, A. J. Kondow, J. Brahm, M. B. Fenderson, G. L. Breisford, and V. B. Pett. 1989. Bioorganic Chemistry. 17:96-107), to approximately 0.001% of control cells, which is a flux as low as in lipid bilayers. The reversible DIDS inhibition of the residual chloride flux depended on the extracellular chloride concentration, but was not purely competitive. The half-inhibition concentrations at [Cl(o)] = 150 mM in control cells (Ki,o) and covalently DIDS-treated cells (Ki,c) were: DIDS, Ki,c = 73 nM; DNDS, Ki,o = 6.3 microM, Ki,c = 22 microM; phloretin, Ki,o = 19 microM, Ki,c = 17 microM; salicylate, Ki,o = 4 mM, Ki,c = 8 mM; Killer III, Ki,o = 10 microM, Ki,c = 10 microM.

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Some effects of low pH on chloride exchange in human red blood cells.

The Journal of General Physiology ◽

10.1085/jgp.65.6.731 ◽

1975 ◽

Vol 65 (6) ◽

pp. 731-749 ◽

Cited By ~ 40

Author(s):

R B Gunn ◽

J O Wieth ◽

D C Tosteson

Keyword(s):

Red Blood Cells ◽

Blood Cells ◽

Chloride Concentration ◽

Low Ph ◽

Chloride Ions ◽

Ph Values ◽

Human Red Blood Cells ◽

Chloride Exchange ◽

Red Cell Membranes ◽

Exchange Flux

In order to test the range of pH values over which the titratable carried model for inorganic anion exchange is valid, chloride self-exchange across human red blood cells was examined between pH 4.75 and 5.7 at 0 decrees c. It was found that chloride self-exchange flux had a minimum near pH 5 and increased again with further increase in hydrogen ion activity. The Arrhenius activation energy for chloride exchange was greatly reduced at low pH values. The chloride flux at pH 5.1 did not show the saturation kinetics reported at higher pH values but was proportional to the value of the chloride concentration squared. In addition, the extent of inhibition of chloride self-exchange flux by phloretin was reduced at low pH. Our interpretation of these findings is that the carrier-mediated flux becomes a progressively smaller fraction of the total flux at lower pH values and that a different transport mode requiring two chloride ions to form the permeant species and having a low specificity and temperature dependence becomes significant below pH5. A possible mechanism for this transport is that chloride crosses red cell membranes as dimers of HCl at these very low pH values.

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