Human fibroblasts bind directly to fibrinogen at RGD sites through integrin alpha(v)beta3.
Fibroblast migration into the blood clot initially filling a wound requires close interaction between fibroblasts and the matrix of the fibrin clot. However, very little is known about the specific receptor-ligand interactions that mediate fibroblast attachment to fibrin. Using an attachment assay developed to measure even relatively weak interactions, we demonstrate here that normal human dermal fibroblasts can attach to substrates coated with fibrinogen, fibrin, or the fibrinogen breakdown product I-9D. Fibroblast attachment to these ligands did not require the presence of fibronectin on the cell surface or as a component of the substrate. Cells treated with cycloheximide and monensin, to limit the synthesis and secretion of endogenous fibronectin, attached as well as untreated cells. The synthetic peptide GRGDS inhibited adhesion to fibrinogen, fibrin, and fibrinogen I-9D by about 60%, while the control peptide GRGES had no substantial effect. We conclude that attachment to these ligands is mediated at least partially by direct interactions between the substrates and one specific receptor, the integrin alpha(v)beta3. Affinity chromatography demonstrated that alpha(v)beta3 from detergent lysates of fibroblasts bound to a fibrinogen matrix and was eluted with EDTA. Furthermore, antibodies against the alpha(v)beta3 complex or against the alpha(v) subunit inhibited fibroblast attachment to fibrinogen and fibrin by 50-70%. An inhibitory antibody against the integrin beta1 subunit had no effect. The observation that integrin antagonists could not produce complete inhibition suggests that there may be other fibroblast cell surface proteins that can bind directly to fibrinogen.
Gailit
J
J
Department of Dermatology, State University of New York at Stony Brook, 11794, USA.
Clarke
C
C
Newman
D
D
Tonnesen
M G
MG
Mosesson
M W
MW
Clark
R A
RA
D002448:Cell Adhesion / Q000502:physiology; D002478:Cells, Cultured; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D006801:Humans; D008297:Male; D009842:Oligopeptides / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D019030:Receptors, Vitronectin / Q000378:metabolism*; D012867:Skin / Q000166:cytology; D014945:Wound Healing
D009842:Oligopeptides; D010446:Peptide Fragments; D019030:Receptors, Vitronectin; C047981:arginyl-glycyl-aspartic acid; D005337:Fibrin; D005340:Fibrinogen