Characterisation of erythrocyte transmembrane exchange of trifluoroacetate using 19F-NMR: evidence for transport via the monocarboxylate transporter.
The transport of trifluoroacetate (TFA) and difluorophosphate (DFP) into and out of human and sheep erythrocytes was measured using 19F-NMR. The pathways for the transport in human erythrocytes were characterised by differentiating between the transport inhibition caused by different reagents. (1) Pre-treatment of human erythrocytes with N-ethylmaleimide (10 mM) caused a decrease of the membrane-permeability coefficients for TFA influx and efflux to 0.74 +/- 0.05 and 0.83 +/- 0.09-times, respectively, of those determined in the absence of inhibition. Concomitantly there was no apparent effect on the band-3-mediated transport of DFP. Thus, the decrease of the permeability of TFA is consistent with the inhibition being that of the monocarboxylate transporter. (2) Inhibition of TFA and DFP exchange was also seen in human erythrocytes treated with p-chloromercuriphenylsulfonate (pCMBS). The extent of inhibition reached a maximum value for the pCMBS concentrations beyond which further inhibition was not achieved and there was substantial residual exchange of the two solutes. (3) Residual flux of TFA was found in the presence of high concentrations of the inhibitors, alpha-cyano-4-hydroxycinnamate (> or = 4 mM) or 4,4'-dinitrostilbene-2,2'-disulfonate (> or = 1 mM) when each compound was used alone. (4) Complete inhibition of TFA uptake was obtained when human erythrocytes were treated with both alpha-cyano-4-hydroxycinnamate (4 mM) and a stilbene disulfonate. It was, therefore, concluded that simple diffusion of TFA via the lipid bilayer was negligible in human erythrocytes and that incomplete inhibition of the monocarboxylate transporter occurred when the compounds were used alone.
Xu
A S
AS
Department of Biochemistry, University of Sydney, NSW, Australia.
Kuchel
P W
PW
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002463:Cell Membrane Permeability; D003373:Coumaric Acids; D004058:Diffusion; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005033:Ethylmaleimide; D005459:Fluorides / Q000378:metabolism; D005461:Fluorine; D006801:Humans; D007773:Lactates / Q000378:metabolism*; D019344:Lactic Acid; D009682:Magnetic Resonance Spectroscopy; D010756:Phosphoric Acids / Q000378:metabolism; D011773:Pyruvates / Q000378:metabolism*; D019289:Pyruvic Acid; D012756:Sheep; D014269:Trifluoroacetic Acid / Q000378:metabolism*
D001457:Anion Exchange Protein 1, Erythrocyte; D003373:Coumaric Acids; D007773:Lactates; D010756:Phosphoric Acids; D011773:Pyruvates; C081381:difluorophosphoric acid; C007175:alpha-cyano-4-hydroxycinnamate; D005461:Fluorine; D019344:Lactic Acid; D019289:Pyruvic Acid; D014269:Trifluoroacetic Acid; D005033:Ethylmaleimide; D005459:Fluorides