Asymmetric and functional reconstitution of band 3 into pre-formed phosphatidylcholine vesicles

Human erythrocyte band 3 protein was purified in 0.1% Triton X-100 and reconstituted into pre-formed phosphatidylcholine vesicles by a Triton X-100-mediated procedure [1]. Band 3 (and its transmembrane domain) could be asymmetrically reconstituted into phosphatidylcholine vesicles with retention of sulfate transport activity which showed behaviour characteristic of red cell anion transport in response to pH, H2DIDS and temperature. Successful reconstitution was also possible using high mol ratios of band 3/phosphatidylcholine (1:200), which are not achieved by any other method.