Dissecting the EphA3/Ephrin-A5 interactions using a novel functional mutagenesis screen Journal Article
| Authors: | Smith, F. M.; Vearing, C.; Lackmann, M.; Treutlein, H.; Himanen, J.; Chen, K.; Saul, A.; Nikolov, D.; Boyd, A. W. |
| Article Title: | Dissecting the EphA3/Ephrin-A5 interactions using a novel functional mutagenesis screen |
| Abstract: | The EphA3 receptor tyrosine kinase preferentially binds ephrin-A5, a member of the corresponding sub-family of membrane-associated ligands. Their interaction regulates critical cell communication functions in normal development and may play a role in neoplasia. Here we describe a random mutagenesis approach, which we employed to study the molecular determinants of the EphA3/ephrin-A5 recognition. Selection and functional characterization of EphA3 point mutants with impaired ephrin-A5 binding from a yeast expression library defined three EphA3 surface areas that are essential for the EphA3/ephrin-A5 interaction. Two of these map to regions identified previously in the crystal structure of the homologous EphB2-ephrin-B2 complex as potential ligand/receptor interfaces. In addition, we identify a third EphA3/ephrin-A5 interface that falls outside the structurally characterized interaction domains. Functional analysis of EphA3 mutants reveals that all three Eph/ephrin contact areas are essential for the assembly of signaling-competent, oligomeric receptor-ligand complexes. |
| Keywords: | human cell; nonhuman; protein domain; protein motif; animal cell; complex formation; protein protein interaction; protein binding; protein tyrosine kinase; animalia; amino acid sequence; cell membranes; crystal structure; models, molecular; dimerization; binding sites; receptor protein-tyrosine kinases; yeast; protein structure; point mutation; protein interaction mapping; genetic screening; biochemistry; mutagenesis; oligomerization; complexation; ephrin-a5; receptor down regulation; cytoarchitecture; chemical bonds; ephrin receptor; ephrin a5; humans; human; priority journal; article; cell communication functions; receptor-ligand complexes; ephrin a3 |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 279 |
| Issue: | 10 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2004-03-05 |
| Start Page: | 9522 |
| End Page: | 9531 |
| Language: | English |
| DOI: | 10.1074/jbc.M309326200 |
| PROVIDER: | scopus |
| PUBMED: | 14660665 |
| DOI/URL: | |
| Notes: | J. Biol. Chem. -- Cited By (since 1996):56 -- Export Date: 16 June 2014 -- CODEN: JBCHA -- Source: Scopus |
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