Biomembrane affinity chromatographic analysis of inhibitor binding to the human red cell nucleoside transporter in immobilized cells, vesicles and proteoliposomes.

The affinity of the human red cell nucleoside transporter for the transport inhibitor nitrobenzylthioinosine decreases upon protein purification. The affinity was highest for the whole cells (Kd, 0.04 nM), lowered upon cytoskeleton depletion (Kd, 0.2 nM) and lowest after partial purification and reconstitution (Kd, 0.3 nM), as determined by frontal affinity chromatography.