Structural recognition of an optimized substrate for the ephrin family of receptor tyrosine kinases.
Ephrin receptor tyrosine kinase A3 (EphA3, EC 2.7.10.1) is a member of a unique branch of the kinome in which downstream signaling occurs in both ligand- and receptor-expressing cells. Consequently, the ephrins and ephrin receptor tyrosine kinases often mediate processes involving cell-cell contact, including cellular adhesion or repulsion, developmental remodeling and neuronal mapping. The receptor is also frequently overexpressed in invasive cancers, including breast, small-cell lung and gastrointestinal cancers. However, little is known about direct substrates of EphA3 kinase and no chemical probes are available. Using a library approach, we found a short peptide sequence that is a good substrate for EphA3 and is suitable for co-crystallization studies. Complex structures show multiple contacts between kinase and substrates; in particular, two residues undergo conformational changes and by mutation are found to be important for substrate binding and turnover. In addition, a difference in catalytic efficiency between EPH kinase family members is observed. These results provide insight into the mechanism of substrate binding to these developmentally integral enzymes.
Davis
Tara L
TL
Structural Genomics Consortium, University of Toronto, Canada.
Walker
John R
JR
Allali-Hassani
Abdellah
A
Parker
Sirlester A
SA
Turk
Benjamin E
BE
Dhe-Paganon
Sirano
S
D018360:Crystallography, X-Ray; D036342:Ephrins / Q000737:chemistry*; D006801:Humans; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D036121:Receptor, EphA3; D013379:Substrate Specificity
D036342:Ephrins; D009363:Neoplasm Proteins; D019151:Peptide Library; C491718:EPHA3 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D036121:Receptor, EphA3
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