Local structural difference between human and bovine band 3 in the anion transport inhibitor-binding region

We have examined molecular properties of inhibitor-complexed human and bovine band 3, an anion transport protein of erythrocyte membrane, in order to demonstrate the structural characteristics of the inhibitor binding region. Band 3 modified with DIDS (4,4'-diisothiocyano-2,2'-stilbenedisulfonate), a potent anion transport inhibitor, generated a positive circular dichroic band at a wavelength of 345 nm, corresponding to a DIDS chromophore. The dichroic spectra of human band 3-DIDS complex and its bovine counterpart differed markedly in their ellipticity. Under the conditions that H2DIDS (the dihydro-derivative of DIDS) cross-linked two chymotryptic fragments of human band 3, the reagent failed to cross-link the equivalent bovine fragments. The inhibitory effect of PLP (pyridoxal 5'-phosphate), a substrate and affinity label, on phosphate influx into red blood cells was more pronounced for human band 3 than for bovine band 3. The residue Lys-562 of human band 3 was found to be modified with PLP, while the corresponding residue of bovine band 3 was devoid of reactivity with PLP.