Nj. Hemming et al., LOCALIZATION OF THE PROTEIN 4.1-BINDING SITE ON HUMAN ERYTHROCYTE GLYCOPHORIN-C AND GLYCOPHORIN-D, Biochemical journal, 299, 1994, pp. 191-196
The flexibility of the human erythrocyte membrane is mediated by an un
derlying network of skeletal proteins which interact with the membrane
through ankyrin and protein 4.1. The nature of the membrane attachmen
t site(s) for protein 4.1 has yet to be fully elucidated. In this pape
r we show that purified protein 4.1 binds much less strongly to alkali
-stripped membranes from erythrocytes of individuals with total glycop
horin C and D deficiency (Leach phenotype) than to alkali-stripped nor
mal membranes. We further show that a synthetic peptide corresponding
to amino acid residues 82-98 of the cytoplasmic domain of glycophorin
C specifically binds to purified protein 4.1 and inhibits protein 4.1
binding to alkali-stripped normal membranes. The same synthetic peptid
e binds directly to membranes from individuals with glycophorin C and
D deficiency but not to normal membranes. These results indicate that
glycophorins C and D provide major membrane attachment sites for prote
in 4.1 in normal erythrocytes and that this interaction is mediated by
protein 4.1 binding to amino acid residues 82-98 of glycophorin C and
61-77 of glycophorin D.