Abstract
The Eph family of receptor tyrosine kinases and their ephrin ligands are mediators of cell-cell communication. Cleavage of ephrin-A2 by the ADAM10 membrane metalloprotease enables contact repulsion between Eph- and ephrin-expressing cells. How ADAM10 interacts with ephrins in a regulated manner to cleave only Eph bound ephrin molecules remains unclear. The structure of ADAM10 disintegrin and cysteine-rich domains and the functional studies presented here define an essential substrate-recognition module for functional interaction of ADAM10 with the ephrin-A5/EphA3 complex. While ADAM10 constitutively associates with EphA3, the formation of a functional EphA3/ephrin-A5 complex creates a new molecular recognition motif for the ADAM10 cysteine-rich domain that positions the proteinase domain for effective ephrin-A5 cleavage. Surprisingly, the cleavage occurs in trans, with ADAM10 and its substrate being on the membranes of opposing cells. Our data suggest a simple mechanism for regulating ADAM10-mediated ephrin proteolysis, which ensures that only Eph bound ephrins are recognized and cleaved.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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ADAM Proteins / chemistry
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ADAM Proteins / genetics
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ADAM Proteins / metabolism*
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ADAM10 Protein
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Amino Acid Sequence
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Amyloid Precursor Protein Secretases
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Blotting, Western
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Cell Line
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Cell Line, Tumor
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Conserved Sequence
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Crystallography, X-Ray
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Cysteine / chemistry
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Disulfides / chemistry
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Ephrin-A2 / metabolism*
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Ephrin-A3 / chemistry
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Ephrin-A3 / metabolism*
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Ephrin-A5 / chemistry
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Ephrin-A5 / metabolism*
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Green Fluorescent Proteins / metabolism
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Humans
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Hydrolysis
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Ligands
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Membrane Proteins / chemistry
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Microscopy, Confocal
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis
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Phylogeny
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Precipitin Tests
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Protein Folding
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Protein Structure, Secondary
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Protein Structure, Tertiary
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RNA Interference
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RNA, Small Interfering / metabolism
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Receptor, EphA3 / metabolism
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Sequence Homology, Amino Acid
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Substrate Specificity
Substances
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Disulfides
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Ephrin-A2
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Ephrin-A3
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Ephrin-A5
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Ligands
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Membrane Proteins
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RNA, Small Interfering
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Green Fluorescent Proteins
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Receptor, EphA3
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Amyloid Precursor Protein Secretases
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ADAM Proteins
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ADAM10 Protein
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ADAM10 protein, human
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Cysteine