Location of a fibronectin domain involved in newt epidermal cell migration.
The interaction of migrating newt epidermal cells with the extracellular matrix protein, fibronectin, was studied. Pieces of nitrocellulose coated with intact human plasma fibronectin or proteolytically derived fragments were implanted into wounded limbs so that the coated nitrocellulose served as wound bed for migrating epidermal cells as they attempted to form a wound epithelium. Epidermal cells migrated very poorly on nitrocellulose pieces coated with (a) a 27-kD amino-terminal heparin-binding fragment, (b) a 46-kD gelatin-binding fragment, (c) a combined 33- and 66-kD carboxy-terminal heparin-binding preparation representing peptide sequences in the A and B chains, respectively, or (d) a 31-kD carboxy-terminal fragment from the A chain, containing a free sulfhydryl group. In contrast, epidermal cells readily migrated onto nitrocellulose coated with a mixture of fragments from the middle of the molecule (80-125kD) that bind neither heparin nor gelatin. Attempts to block migration on fibronectin-coated nitrocellulose using IB10, a monoclonal antibody that blocks Chinese hamster ovary cell attachment to fibronectin, were unsuccessful despite saturation of the epitope against which IB10 is directed. In contrast, a polyclonal anti-fibronectin antibody did inhibit migration. These results show that the ability of fibronectin to support newt epidermal cell migration is not shared equally by all regions of the molecule, but is restricted to a domain in the middle third. They also suggest that the site supporting migration is separate and distinct from the site mediating Chinese hamster ovary cell attachment.
D000818:Animals; D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D002465:Cell Movement; D003101:Collodion; D000078404:Epidermal Cells; D005109:Extracellular Matrix / Q000502:physiology; D005353:Fibronectins / Q000276:immunology / Q000502:physiology*; D008297:Male; D010446:Peptide Fragments; D011487:Protein Conformation; D012455:Salamandridae; D013329:Structure-Activity Relationship; D014945:Wound Healing
D000911:Antibodies, Monoclonal; D005353:Fibronectins; D010446:Peptide Fragments; D003101:Collodion