Marfatia, S. M., Morais-Cabral, J. H., Kim, A. C., Byron, O. and Chishti, A. H. (1997) The PDZ domain of human erythrocyte p55 mediates its binding to the cytoplasmic carboxyl terminus of glycophorin C: analysis of the binding interface by in vitromutagenesis. Journal of Biological Chemistry, 272(39), pp. 24191-24197. (doi: 10.1074/jbc.272.39.24191) (PMID:9305870)
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Abstract
The PDZ domain, also known as the GLGF repeat/DHR domain, is an ∼90-amino acid motif discovered in a recently identified family of proteins termed MAGUKs (membrane-associated guanylatekinase homologues). Sequence comparison analysis has since identified PDZ domains in over 50 proteins. Like SH2 and SH3 domains, the PDZ domains mediate specific protein-protein interactions, whose specificities appear to be dictated by the primary structure of the PDZ domain as well as its binding target. Using recombinant fusion proteins and a blot overlay assay, we show that a single copy of the PDZ domain in human erythrocyte p55 binds to the carboxyl terminus of the cytoplasmic domain of human erythroid glycophorin C. Deletion mutagenesis of 21 amino acids at the amino terminus of the p55 PDZ domain completely abrogates its binding activity for glycophorin C. Using an alanine scan and surface plasmon resonance technique, we identify residues in the cytoplasmic domain of glycophorin C that are critical for its interaction with the PDZ domain. The recognition specificity of the p55 PDZ domain appears to be unique, since the three PDZ domains of hDlg (human lymphocyte homologue of the Drosophiladiscs large tumor suppressor) do not bind the cytoplasmic domain of glycophorin C. Taken together with our previous studies, these results complete the identification of interacting domains in the ternary complex between p55, glycophorin C, and protein 4.1. Implications of these findings are discussed in terms of binding specificity and the regulation of cytoskeleton-membrane interactions.
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Byron, Professor Olwyn |
Authors: | Marfatia, S. M., Morais-Cabral, J. H., Kim, A. C., Byron, O., and Chishti, A. H. |
College/School: | College of Medical Veterinary and Life Sciences |
Journal Name: | Journal of Biological Chemistry |
ISSN: | 0021-9258 |
ISSN (Online): | 1083-351X |
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