The Carboxyl Terminus of B Class Ephrins Constitutes a PDZ Domain Binding Motif*
- From the ‡Programme in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario M5G 1X5, Canada, the§Department of Molecular and Medical Genetics, University of Toronto, Toronto, Ontario M5S 1A8, Canada, and the ‖Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139
Abstract
Ephrin B proteins function as ligands for B class Eph receptor tyrosine kinases and are postulated to possess an intrinsic signaling function. The sequence at the carboxyl terminus of B-type ephrins contains a putative PDZ binding site, providing a possible mechanism through which transmembrane ephrins might interact with cytoplasmic proteins. To test this notion, a day 10.5 mouse embryonic expression library was screened with a biotinylated peptide corresponding to the carboxyl terminus of ephrin B3. Three of the positive cDNAs encoded polypeptides with multiple PDZ domains, representing fragments of the molecule GRIP, the protein syntenin, and PHIP, a novel PDZ domain-containing protein related toCaenorhabditis elegans PAR-3. In addition, the binding specificities of PDZ domains previously predicted by an oriented library approach (Songyang, Z., Fanning, A. S., Fu, C., Xu, J., Marfatia, S. M., Chishti, A. H., Crompton, A., Chan, A. C., Anderson, J. M., and Cantley, L. C. (1997)Science 275, 73–77) identified the tyrosine phosphatase FAP-1 as a potential binding partner for B ephrins. In vitro studies demonstrated that the fifth PDZ domain of FAP-1 and full-length syntenin bound ephrin B1 via the carboxyl-terminal motif. Lastly, syntenin and ephrin B1 could be co-immunoprecipitated from transfected COS-1 cells, suggesting that PDZ domain binding of B ephrins can occur in cells. These results indicate that the carboxyl-terminal motif of B ephrins provides a binding site for specific PDZ domain-containing proteins, which might localize the transmembrane ligands for interactions with Eph receptors or participate in signaling within ephrin B-expressing cells.
Footnotes
-
↵* This work was supported by the Medical Research Council (MRC) of Canada and by a Howard Hughes Medical Institute International Research Scholar Award (to T. P.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
-
↵¶ Supported by a graduate studentship from the MRC of Canada.
-
↵** Distinguished Scientist of the MRC of Canada. To whom correspondence should be addressed. Tel.: 416-586-8262; Fax: 416-586-8857; E-mail: pawson{at}mshri.on.ca.
-
↵2 G. Mbamalu, S. Holland, and T. Pawson, unpublished results.
-
↵3 Z. Songyang, unpublished results.
- Abbreviations:
- PDZ
-
PSD95/Dlg, ZO-1
- GST
-
glutathione S-transferase
- FAP
-
Fas-associated phosphatase.
-
- Received June 16, 1998.
- Revision received October 15, 1998.
- The American Society for Biochemistry and Molecular Biology, Inc.
