title: Mapping of ankyrin binding determinants on the erythroid anion exchanger, AE1.
abstract: The association of ankyrin with the AE1 anion exchanger contributes an essential function to the mechanical and viscoelastic properties of the erythrocyte and constitutes the best understood link between the plasma membrane and the underlying membrane skeleton. The AE1 binding domain of ankyrin consists of 24 tandem repeats of a 33-amino acid motif that is present on a wide variety of otherwise unrelated proteins. The experiments described in this paper are aimed at identifying the specific amino acid sequences in AE1 that comprise the ankyrin binding site. We have exploited a cell-free binding assay to quantify the binding affinity of anion exchangers and a recombinant fragment of ANK1, R13-H. Our previous study (Ding, Y., Casey, J. R. and Kopito, R. R. (1995) J. Biol. Chem. 269, 32201-32208) identified an essential role of the amino-terminal 79 AE1 residues in ankyrin binding. The present study extends these findings to show that these 79 amino acids, although necessary, are not sufficient for ankyrin binding. Using chimeras between AE1 and the closely related anion exchanger AE2, which does not bind ankyrin, we have defined a 40-residue region of AE1 between positions 155 and 195 that is also essential for ankyrin binding.
authors.affiliation: Department of Biological Sciences, Stanford University, Stanford, California 94305-5020, USA.
authors.forename: Y
authors.initials: Y
authors.lastname: Ding
authors.forename: S
authors.initials: S
authors.lastname: Kobayashi
authors.forename: R
authors.initials: R
authors.lastname: Kopito
mesh_terms: D001457:Anion Exchange Protein 1, Erythrocyte / Q000378:metabolism*; D017487:Ankyrins / Q000378:metabolism*; D001665:Binding Sites; D002460:Cell Line; D006801:Humans; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D001457:Anion Exchange Protein 1, Erythrocyte; D017487:Ankyrins; D011993:Recombinant Fusion Proteins