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.
Haneskog
L
L
Department of Biochemistry, Biomedical Center, Uppsala University, Box 576, S-751 23 Uppsala, Sweden.
Zeng
C M
CM
Lundqvist
A
A
Lundahl
P
P
D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods*; D003571:Cytochalasin B / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000737:chemistry; D004912:Erythrocytes / Q000737:chemistry*; D051272:Glucose Transporter Type 1; D006801:Humans; D007700:Kinetics; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D009004:Monosaccharide Transport Proteins / Q000737:chemistry; D033703:Nucleoside Transport Proteins; D011510:Proteolipids / Q000737:chemistry*; D013868:Thioinosine / Q000031:analogs & derivatives* / Q000737:chemistry
D002352:Carrier Proteins; D051272:Glucose Transporter Type 1; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D033703:Nucleoside Transport Proteins; D011510:Proteolipids; C495677:SLC2A1 protein, human; C020485:proteoliposomes; D003571:Cytochalasin B; D013868:Thioinosine; C001789:4-nitrobenzylthioinosine