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Blood, Vol. 92 No. 11 (December 1), 1998:
pp. 4428-4438
Functional Cell Surface Expression of Band 3, the Human Red
Blood Cell Anion Exchange Protein (AE1), in K562
Erythroleukemia Cells: Band 3 Enhances the Cell Surface Reactivity
of Rh Antigens
Roland Beckmann,
Jonathan S. Smythe,
David J. Anstee, and
Michael
J.A. Tanner
From the Department of Biochemistry, University of Bristol, UK; and
the International Blood Group Reference Laboratory, Bristol, UK.
Human K562 erythroleukemia cells were transfected with human band 3 (anion exchanger 1 [AE1]) cDNA, using the pBabe
retroviral vector. Stable K562 clones expressing band 3 were isolated
by flow cytometry, and surface expression was quantified by
immunoblotting. The function of band 3 expressed at the cell surface
was demonstrated in chloride transport assays. K562 cells expressing
band 3 also displayed high levels of the Wrb blood group
antigen, confirming the role of band 3 in Wrb expression,
and an increase in the low levels of endogenous Rh antigen activity. We
also performed coexpression experiments with K562 clones that had
previously been transduced with cDNAs encoding RhD or RhcE
polypeptides. The transfection and expression of band 3 in these clones
substantially increased the levels of RhD and cE antigen activity
expressed on the cells and also increased the reactivity of the cells
with antibody to the endogenous Rh glycoprotein (RhGP, Rh50). The
increased reactivity of Rh antigens may result from cell surface or
intracellular interactions of band 3 with the protein complex which
contains the Rh polypeptides and RhGP, or from indirect effects of band
3 on the membrane environment. This work establishes a system for cell
surface expression of band 3 in a mammalian cell line, which will
enable further studies of the protein and its interactions with other
membrane components.
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