title: Reconstitution of glucose transport using human erythrocyte band 3.
abstract: Band 3 and the diffuse component of zone 4.5, designated band 4.5.B, have been separately prepared from human erythrocyte membranes and incorporated into the membranes of 150 nm 1-palmitoyl-2-oleoyl phosphatidylcholine vesicles. The rates of glucose influx into these vesicles were measured under zero-trans conditions. Both sets of vesicles exhibited substrate-saturable transport which was inhibited by phloretin. However, the specific activity of the band 3 vesicles, 292 mumol X min-1 X (mg protein)-1, was more than twice that of the band 4.5.B vesicles, and the turnover number of transporters in the band 3 vesicles was at least 4-fold greater than those in the 4.5.B vesicles. Very little background density was visible in the band 4.5 region of erythrocyte membranes protected from degradation. In unprotected membranes, band 4.5.B was abundantly present, could be purified, and had glucose transport activity. Previously we have shown (Biochemistry 19, 1205 (1980] that maltosyl isothiocyanate, an affinity label for the glucose transporter, labelled a single 100 000 Mr protein of the intact erythrocyte membrane. Based upon the results of both affinity labelling and reconstitution we suggest that the native glucose transporter is a component of band 3, and that band 4.5.B contains a partially active fragment of the native transporter.
mesh_terms: D001457:Anion Exchange Protein 1, Erythrocyte; D001786:Blood Glucose / Q000378:metabolism*; D001798:Blood Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009004:Monosaccharide Transport Proteins
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D001786:Blood Glucose; D001798:Blood Proteins; D002352:Carrier Proteins; D009004:Monosaccharide Transport Proteins