Red-cell glycophorin A-band 3 interactions associated with the movement of band 3 to the cell surface.
We have examined the mechanism by which glycophorin A (GPA) facilitates the movement of the human red-cell anion exchanger (band 3, AE1) to the cell surface. GPA itself forms stable dimers in membranes and detergent solution. Four mutants of human GPA with impaired dimerization were prepared (L75I, I76A, G79L and G83L). All four GPA mutants enhanced band 3 translocation to the Xenopus oocyte plasma membrane in the same way as wild-type GPA, showing that the GPA monomer is sufficient to mediate this process. Cell-surface expression of the natural band 3 mutant G701D has an absolute requirement for GPA. GPA monomers also rescued the cell-surface expression of this mutant band 3. Taking into account other evidence, we infer that the site of GPA interaction with band 3 is located outside the GPA dimerization interface but within the GPA transmembrane span. The results of examination of the cell-surface expression of GPA and band 3 in different K562 erythroleukaemia cell clones stably transfected with band 3 are consistent with the movement of GPA and band 3 to the cell surface together. We discuss the pathways by which band 3 moves to the cell surface in the presence and the absence of GPA, concluding that GPA has a role in enhancing the folding and maturation of band 3. We propose that GPA functions in erythroid cells to assist with the incorporation of large amounts of properly folded band 3 into the membrane within a limited time span during erythroid maturation.
Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, U.K.
M T
MT
Young
R
R
Beckmann
A M
AM
Toye
M J
MJ
Tanner
D000818:Animals; D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers; D019281:Dimerization; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism*; D006021:Glycophorins / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D021381:Protein Transport; D014981:Xenopus
D001457:Anion Exchange Protein 1, Erythrocyte; D017931:DNA Primers; D006021:Glycophorins