Modification by papain of the structure and function of band 3, the erythrocyte anion transport protein.
Extracellular papain is known to inhibit the anion transport function of the band 3 protein of the human red blood cell membrane. Previous work [Jennings, M. L., & Passow, H. (1979) Biochim. Biophys. Acta 554, 498-519] had suggested that this inhibition may result from the removal by papain of 5 000-10 000 daltons from the 35 000-dalton chymotryptic peptide of band 3. The present work shows, however, that papain also removes a small peptide from the C terminus of the 60 000-dalton chymotryptic peptide. The C-terminal amino acid sequence of this peptide is -Lys-Thr-Tyr. Whether or not this newly discovered action of papain is responsible for inhibiting anion transport is unknown. The effects of extracellular papain on the band 3 function have been characterized in detail. Papain inhibits Cl-Cl exchange in a high Cl medium by almost 90%. This inhibition appears to result from inhibition of the efflux step in the catalytic cycle for the transport, because papain does not inhibit the anion transport when it is assayed under influx-limited conditions. Moreover, since papain has no detectable effect on the dissociation constant for extracellular substrate (SO4) binding, the material removed by papain cannot be involved closely in the outward-facing substrate site. In contrast, removal of this material strongly (12-fold) reduces the affinity of the inhibitor 4,4'-dinitro-2,2'-stilbenedisulfonate for outward-facing sites. Therefore, stilbenedisulfonate binding involves portions of the band 3 molecule which are not intimately related to substrate binding.
D000328:Adult; D001457:Anion Exchange Protein 1, Erythrocyte; D001693:Biological Transport, Active; D001798:Blood Proteins / Q000378:metabolism*; D002712:Chlorides / Q000097:blood; D002918:Chymotrypsin; D006801:Humans; D007700:Kinetics; D010206:Papain / Q000494:pharmacology*; D010446:Peptide Fragments
D001457:Anion Exchange Protein 1, Erythrocyte; D001798:Blood Proteins; D002712:Chlorides; D010446:Peptide Fragments; D002918:Chymotrypsin; D010206:Papain