Monoclonal antibodies to the glucose transporter from human erythrocytes. Identification of the transporter as a Mr = 55,000 protein.
Using the preparation of purified glucose transporter from human erythrocytes as antigen, we have prepared and characterized six monoclonal antibodies. Three of these antibodies have been shown to be to the glucose transporter by several criteria: they immunoprecipitate the transport activity, the cytochalasin B binding activity, and 75% of the protein from the solubilized purified preparation. The remaining three antibodies were shown to recognize the same polypeptide by a Western blot procedure. All of the antibodies reacted with the deglycosylated transporter and are thus against peptide determinants; most bound to the cytoplasmic domain of the transporter. The antibodies exhibited a range of effects on cytochalasin B binding, from slight enhancement to modest inhibition to strong inhibition; for this reason they must bind to at least three different epitopes. Western blot analysis of erythrocyte membranes prepared in the presence of protease inhibitors showed that all six antibodies bound to a polypeptide of average Mr = 55,000. Moreover, by immunological assay this polypeptide accounted for 5.3% of the membranes protein, a value similar to that given by cytochalasin B binding. Thus, the proposal that the native transporter is a Mr = 100,000 polypeptide is highly unlikely. The antibodies also react with the glucose transporter in other human cell types, but not with that in rodent or avian cells.
W J
WJ
Allard
G E
GE
Lienhard
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001665:Binding Sites; D001786:Blood Glucose / Q000378:metabolism*; D002352:Carrier Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D002642:Chick Embryo; D003571:Cytochalasin B / Q000378:metabolism; D000939:Epitopes / Q000032:analysis; D004910:Erythrocyte Membrane / Q000032:analysis; D004912:Erythrocytes / Q000032:analysis*; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins; D009005:Monosaccharides / Q000097:blood*; D051381:Rats
D000911:Antibodies, Monoclonal; D001786:Blood Glucose; D002352:Carrier Proteins; D000939:Epitopes; D009004:Monosaccharide Transport Proteins; D009005:Monosaccharides; D003571:Cytochalasin B