Interaction between anions and the reduced folate/methotrexate transport system in L1210 cell plasma membrane vesicles: directional symmetry and anion specificity for differential mobility of loaded and unloaded carrier.

The effect of various anions on the mediated influx and efflux of [3H]methotrexate by L1210 cell plasma membrane vesicles in a HEPES buffer system was studied. Our results show that flux is stimulated to the same extent in either direction when SO4, Pi, or folate compounds (1,L5-CHO-folate-H4, methotrexate), but not Cl- was present in the opposite compartment. This implies the property of directional symmetry, a condition in which differential mobility of loaded and unloaded carriers occurs in both directions. We also observed a similarity in the specificity of the interaction between various anions and carrier in each orientation of the membrane, in the order, Cl- much less than Pi approximately equal to SO2-4 much less than methotrexate less than 1,L5-CHO-folate-H4. Also, the absolute differential in mobility of loaded and unloaded carrier (assumed from the extent of transstimulation obtained) varied substantially among the anions examined. No stimulation was obtained with Cl-, and stimulation was twofold with Pi, SO2-4 and methotrexate and fourfold with 1,L5-CHO-folate-H4. Transstimulation of flux from either external or internal compartment only occurred when a positive gradient of total anions was maintained in the opposite compartment. Also, no stimulation occurred when the same equivalence of two different anions are present in opposing compartments. The concentration of anions required to transstimulate [3H]methotrexate influx was increased four- to 10-fold when vesicles were equilibrated in 145 mM NaCl.(ABSTRACT TRUNCATED AT 250 WORDS)