Three distinct molecular surfaces in Ephrin-A5 are essential for a functional interaction with EphA3 Journal Article
| Authors: | Day, B.; To, C.; Himanen, J. P.; Smith, F. M.; Nikolov, D. B.; Boyd, A. W.; Lackmann, M. |
| Article Title: | Three distinct molecular surfaces in Ephrin-A5 are essential for a functional interaction with EphA3 |
| Abstract: | Eph receptor tyrosine kinases (Ephs) function as molecular relays that interact with cell surface-bound ephrin ligands to direct the position of migrating cells. Structural studies revealed that, through two distinct contact surfaces on opposite sites of each protein, Eph and ephrin binding domains assemble into symmetric, circular heterotetramers. However, Eph signal initiation requires the assembly of higher order oligomers, suggesting additional points of contact. By screening a random library of EphA3 binding-compromised ephrin-A5 mutants, we have now determined ephrin-A5 residues that are essential for the assembly of high affinity EphA3 signaling complexes. In addition to the two interfaces predicted from the crystal structure of the homologous EphB2-ephrin-B2 complex, we identified a cluster of 10 residues on the ephrin-A5 E α-helix, the E-F loop, the underlying H β-strand, as well as the nearby B-C loop, which define a distinct third surface required for oligomerization and activation of EphA3 signaling. Together with a corresponding third surface region identified recently outside of the minimal ephrin binding domain of EphA3, our findings provide experimental evidence for the essential contribution of three distinct protein-interaction interfaces to assemble functional EphAS signaling complexes. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc. |
| Keywords: | signal transduction; controlled study; human cell; mutation; nonhuman; mutant protein; binding affinity; protein domain; proteins; animal cell; complex formation; protein assembly; protein protein interaction; cell line; protein binding; enzyme activity; gene library; transfection; protein tyrosine kinase; time factors; blotting, western; amino acid sequence; molecular sequence data; sequence homology, amino acid; kinetics; recombinant fusion proteins; nucleotide sequence; ligand; immunoprecipitation; cell migration; protein induction; binding site; crystal structure; models, molecular; mutagenesis, site-directed; protein structure, tertiary; binding energy; protein structure; sequence homology; point mutation; structure analysis; surface plasmon resonance; ephrin receptor b2; mutagenesis; oligomerization; amino acid motifs; tetramer; ephrin; cells; ephrin b2; ephrin-a5; alpha helix; surface property; oligomers; ephrin receptor; oligomer; beta sheet; ephrin a5; receptor, epha3; ephrin receptor a3; molecular relays; molecular surfaces; signalling complexes |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 280 |
| Issue: | 28 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 2005-07-15 |
| Start Page: | 26526 |
| End Page: | 26532 |
| Language: | English |
| DOI: | 10.1074/jbc.M504972200 |
| PUBMED: | 15901737 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 20" - "Export Date: 24 October 2012" - "CODEN: JBCHA" - "Molecular Sequence Numbers: GENBANK: NM_001962;" - "Source: Scopus" |
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