The effect of carboxymethylating a single methionine residue on the subunit interactions of glycophorin A.
Human red cell glycophorin A shows an equilibrium between dimeric and monomeric forms which have been disignated PAS-1 and PAS-2, respectively. This equilibrium, which is dependent upon protein concentration is achieved by incubation in sodium dodecyl sulfate solutions at elevated temperatures and is assayed by sodium dodecyl sulfate gel electrophoresis. Carboxymethylation of glycophorin A in guanidine hydrochloride or urea alters the interactions between polypeptide chains so that the lower molecular weight form (PAS-2) is obtained much more readily. If the carboxymethylation is performed at pH 3.0 the reaction is limited to the two methionine residues of glycophorin A which are located at positions 8 and 81 in the sequence. In the presence of sodium dodecyl sulfate, only one of the two methionine residues is carboxymethylated, and glycoprotein modified under these conditions does not exhibit the change in electrophoretic mobility. Experiments with [1-14C]iodoacetic acid demonstrated that Met-81, located in the hydrophobic domain of the protein, is the residue protected by sodium dodecyl sulfate. Modification of Met-81 destabilizes the dimeric form relative to the monomer by weakening the interactions between polypeptide chains. The experiments described in this paper confirm that the hydrophobic domain of glycophorin A is involved in subunit interactions and that Met-81 plays a critical role in those interactions.
D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism; D006023:Glycoproteins / Q000097:blood*; D006146:Guanidines; D006801:Humans; D007461:Iodoacetates; D046911:Macromolecular Substances; D008715:Methionine / Q000032:analysis; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation
D000596:Amino Acids; D006023:Glycoproteins; D006146:Guanidines; D007461:Iodoacetates; D046911:Macromolecular Substances; D008715:Methionine