Interactions of the chelating agent 2,3-dimercaptopropane-1-sulfonate with red blood cells in vitro. II. Effects on metalloproteins.
The implications of the carrier mediated uptake of 2,3-dimercaptopropane-1-sulfonate (DMPS) (D.B. Wildenauer et al., Chem.-Biol. Interact., 42 (1982) 165) on cytoplasmic components of human red blood cells have been investigated in vitro. The water-soluble chelating agent caused a mobilization of metals (zinc and copper) from metalloproteins which resulted in a permeation of the membrane. Furthermore, a cytoplasmic protein was found to be attached to the membrane after DMPS treatment of red blood cells. The protein was isolated and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), amino acid analysis and finger-printing as carbonic anhydrase. The enzyme could be solubilized from the membrane by addition of beta-mercaptoethanol, suggesting an involvement of sulfhydryl-groups. In a reconstitution experiment, DMPS-treated human carbonic anhydrase could be attached to inside-out vesicles which were prepared from human erythrocytes. In contrast, bovine carbonic anhydrase, which is known to lack sulfhydryl-groups, failed to bind to the same vesicles. Moreover, attachment of carbonic anhydrase to the membrane did not occur when intact bovine erythrocytes were treated with DMPS. It is suggested that zinc-depletion of carbonic anhydrase causes the liberation of a sulfhydryl-group of the enzyme. This is followed by a disulfide formation with a component of the membrane which results in the observed membrane attachment.
D000596:Amino Acids / Q000032:analysis; D002250:Carbon Radioisotopes; D002256:Carbonic Anhydrases / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D004112:Dimercaprol / Q000031:analogs & derivatives*; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000032:analysis; D014494:Unithiol / Q000378:metabolism / Q000494:pharmacology*; D015032:Zinc / Q000378:metabolism*
D000596:Amino Acids; D002250:Carbon Radioisotopes; D008565:Membrane Proteins; D004112:Dimercaprol; D014494:Unithiol; D003300:Copper; D002256:Carbonic Anhydrases; D015032:Zinc