Distribution and function of multiple anion transporter proteins in brain tumor cell lines in relation to glucose...

The anion transport, "band 3," family of proteins in mammalian brain performs the same functions as that of erythroid band 3. These functions are anion transport, ankyrin binding, and generation of senescent cell antigen, an aging antigen that terminates the life of cells. The presence of 5-7 band 3 related proteins in brain tissue was suggested by the reaction of antibodies to synthetic peptides of erythroid band 3 with a number of bands in immunoblots. Since there are a number of different cell types in brain, tissue cultures of neural cell types were examined to determine whether multiple band 3 related proteins are present in each cell type or whether several band 3 related proteins are present in each cell type. The tumor cell lines exhibit anion transport and are inhibited by the anion transport inhibitors 4,4'-diisothiocyano-2,2' disulfonic acid (DIDS), phenylglyoxal, and furosemide. Glucose transport is inhibited by cytochalasin B and the anion transport inhibitor, phenylglyoxal, in these cell lines, but not by 4,4'-diisothiocyano-2,2' disulfonic acid. Furosemide gave partial inhibition of most, but not all, cell lines. Since phenylglyoxal inhibits anion transport by binding to an arginine near the transport site, inhibition of glucose transport by phenylglyoxal suggests that an arginine lies in the substrate binding site. The number of cytochalasin B and DIDS binding sites was quantitated on cell lines as an approximation of the number of glucose transporter and anion transporter sites, respectively.