ADAM13 disintegrin and cysteine-rich domains bind to the second heparin-binding domain of fibronectin.
ADAM13 is a member of the disintegrin and metalloprotease protein family that is expressed on cranial neural crest cells surface and is essential for their migration. ADAM13 is an active protease that can cleave fibronectin in vitro and remodel a fibronectin substrate in vivo. Using a recombinant secreted protein containing both disintegrin and cysteine-rich domains of ADAM13, we show that this "adhesive" region of the protein binds directly to fibronectin. Fibronectin fusion proteins corresponding to the various functional domains were used to define the second heparin-binding domain as the ADAM13 binding site. Mutation of the syndecan-binding site (PPRR --> PPTM) within this domain abolishes binding of the recombinant disintegrin and cysteine-rich domains of ADAM13. We further show that the adhesive disintegrin and cysteine-rich domain of ADAM13 can promote cell adhesion via beta(1) integrins. This adhesion requires integrin activation and can be prevented by antibodies to the cysteine-rich domain of ADAM13 and beta(1) integrin. Finally, wild type, but not the E/A mutant of ADAM13 metalloprotease domain, can be shed from the cell surface, releasing the metalloprotease domain associated with the disintegrin and cysteine-rich domains. This suggests that ADAM13 shedding may involve its own metalloprotease activity and that the released protease may interact with both integrins and extracellular matrix proteins.
Gaultier
Alban
A
Department of Cell Biology, University of Virginia School of Medicine, Charlottesville, Virginia 22908, USA.
Cousin
Hélène
H
Darribère
Thierry
T
Alfandari
Dominique
D
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002465:Cell Movement; D003545:Cysteine; D019483:Disintegrins / Q000378:metabolism*; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D019012:Integrin beta1 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D008666:Metalloendopeptidases / Q000378:metabolism*; D011509:Proteoglycans / Q000378:metabolism; D053667:Syndecans
D019483:Disintegrins; D005353:Fibronectins; D019012:Integrin beta1; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D053667:Syndecans; D006493:Heparin; D008666:Metalloendopeptidases; D003545:Cysteine