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Biomembrane affinity chromatographic analysis of nitrobenzylthioinosine binding to the reconstituted human red cell nucleoside transporter
Author(s) -
Haneskog Lars,
Lundqvist Andreas,
Lundahl Per
Publication year - 1998
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/(sici)1099-1352(199812)11:1/6<58::aid-jmr390>3.0.co;2-s
Subject(s) - nucleoside , nucleoside transporter , chemistry , transporter , red blood cell , biological membrane , chromatography , affinity chromatography , membrane , biochemistry , enzyme , gene
Solute interactions with membrane proteins can be analyzed by biomembrane affinity chromatography (BAC), previously applied to the human red cell glucose transporter. As a novel example, frontal BAC analysis of interactions between the nucleoside transport inhibitor nitrobenzylthioinosine (NBTI) and immobilized reconstituted nucleoside and glucose transporters from human red cells revealed two binding sites, presumably corresponding to the two transporters. The affinities and amounts of sites were determined by use of a double rectangular hyperbolic equation. The K d value for NBTI binding to the nucleoside transporter in egg phospholipid proteoliposomes was 0.38 ± 0.08 nM (22 °C, I = 0.16, pH 7.4), lower than previously reported for reconstituted systems. The molar ratio between the amounts of nucleoside transporter sites for NBTI and glucose transporter sites for cytochalasin B was 4.5 ± 0.6%. Copyright © 1998 John Wiley & Sons, Ltd.