Rabbit corneal epithelial cells adhere to two distinct heparin-binding synthetic peptides derived from fibronectin.
Fibronectin plays an important role in corneal reepithelialization during corneal wound healing. In this study, rabbit corneal epithelial (RCE) cell adhesion to fibronectin was further defined using proteolytic fragments of fibronectin and chemically synthesized peptides derived from the amino acid sequence of fibronectin. RCE cells adhere to intact fibronectin, the 75 kD fragment containing the RGDS (Arg-Gly-Asp-Ser) cell adhesion-promoting sequence, and the 33/66 kD cell adhesion promoting/heparin-binding fragments of fibronectin. The 75 kD fragment and the 33/66 kD fragments partially inhibited RCE cell adhesion to intact fibronectin, suggesting that these fragments represent distinct sites used by RCE cells to adhere to intact fibronectin. Two chemically synthesized peptides derived from the amino acid sequence of the 33/66 kD fragments of fibronectin, FN-C/H-I (YEKPGSPPREVVPRPRPGV) and FN-C/H-III (YRVRVTPKEKTGPMKE), directly promoted the adhesion of RCE cells. As further evidence that FN-C/H-I and FN-C/H-III play a role in the adhesion of RCE cells to the 33/66 kD fragments of fibronectin, we have shown that soluble FN-C/H-I and FN-C/H-III inhibited RCE cell adhesion on surfaces coated with the 33/66 kD fragments. In addition, polyclonal IgG against FN-C/H-I and FN-C/H-III partially blocked RCE cell adhesion to the 33/66 kD fragments, confirming that these sequences represent adhesion-promoting sites within these fragments. In contrast, two previously described peptides from the 33/66 kD fragments of fibronectin, which promoted the adhesion of a variety of cell types, FN-C/H-II (KNNQKSEPLIGRKKT) and CS-1 (DELPQLVTLPHPNLHG-PEILDVPST), did not support RCE cell adhesion.(ABSTRACT TRUNCATED AT 250 WORDS)
Mooradian
D L
DL
Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455.
McCarthy
J B
JB
Cameron
J D
JD
Skubitz
A P
AP
Furcht
L T
LT
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002448:Cell Adhesion; D002478:Cells, Cultured; D003315:Cornea / Q000378:metabolism / Q000502:physiology*; D004848:Epithelium / Q000378:metabolism / Q000502:physiology; D005353:Fibronectins / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D006493:Heparin / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism / Q000502:physiology*; D011817:Rabbits
D005353:Fibronectins; D007074:Immunoglobulin G; D010446:Peptide Fragments; D006493:Heparin