Human erythrocyte band 3 polymorphism (band 3 Memphis): characterization of the structural modification (Lys 56----Glu) by protein chemistry methods.
Band 3 variants occur rather frequently in different populations. Based on sodium dodecyl sulfate (SDS)-polyacrylamide electrophoretic properties, a widespread polymorphism (band 3 Memphis) has been previously described. It corresponds to a protein that has been hypothesized to be elongated in its N-terminal cytoplasmic domain. Band 3 from a heterozygote subject for this polymorphism and that displays a normal reactivity towards stilbene disulfonates has been isolated and its primary structure determined by protein chemistry. Reverse-phase high performance liquid chromatography tryptic peptide mapping showed, as the only difference with controls, that the enzymatic cleavage between the two N-terminal peptides did not occur, yielding a 69 residue-long fragment. Further cleavages of this peptide (cyanogen bromide, V8 protease), amino acid composition, and sequence analyses demonstrated that the lysine at position 56 was replaced by a glutamic acid. Thus, surprisingly, a single amino acid change is responsible for the large difference in the electrophoretic behavior. This result suggests that single amino acid substitutions may similarly be involved in the structural modification of several other protein variants, described as elongated or shortened based only on SDS-polyacrylamide electrophoresis studies. When deletions/insertions were confirmed by sequence analysis, their extent was often different from that expected from electrophoresis.
Unité INSERM 299, Hôpital de Bicêtre, Le Kremlin-Bicêtre, France.
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D
Yannoukakos
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Vasseur
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Driancourt
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Blouquit
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Delaunay
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Wajcman
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Bursaux
D001457:Anion Exchange Protein 1, Erythrocyte / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D003488:Cyanogen Bromide; D004591:Electrophoresis, Polyacrylamide Gel; D005971:Glutamates; D018698:Glutamic Acid; D006579:Heterozygote; D006801:Humans; D008239:Lysine; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D011110:Polymorphism, Genetic; D012697:Serine Endopeptidases / Q000378:metabolism; D013267:Stilbenes / Q000378:metabolism; D014357:Trypsin / Q000378:metabolism
D001457:Anion Exchange Protein 1, Erythrocyte; D005971:Glutamates; D010446:Peptide Fragments; D013267:Stilbenes; C070316:band 3 protein Memphis; C006711:benzoylamido-4'-aminostilbene-2,2'-disulfonate; D018698:Glutamic Acid; D012697:Serine Endopeptidases; D002918:Chymotrypsin; C028344:glutamyl endopeptidase; D014357:Trypsin; D008239:Lysine; D003488:Cyanogen Bromide