Fibroblasts spread on immobilized fibrin monomer by mobilizing a beta1-class integrin, together with a vitronectin receptor alphavbeta3 on their surface.
Human and murine fibroblasts were found to spread far more avidly on fibrin monomer monolayers than on immobilized fibrinogen, indicating that removal of fibrinopeptides by thrombin is a prerequisite for the fibrin-mediated augmentation of cell spreading. In fact, cell spreading was not efficiently augmented on monolayers of a thrombin-treated dysfibrinogen lacking the release of fibrinopeptide A due to an Aalpha Arg-16 --> Cys substitution. Since a synthetic Arg-Gly-Asp (RGD)-containing peptide inhibited the fibrin-mediated cell spreading, subsequent dissociation of the carboxyl-terminal globular domain of the Aalpha-chains appears to render the RGD segments accessible to the cell-surface integrins. In support of this, fibrin-augmented cell spreading was inhibited by an antibody recognizing a 12-kDa peptide segment with gamma Met-89 at its amino terminus, which is located in close association with the RGD segment at Aalpha 95-97 in the helical coiled-coil interdomainal connector. The fibrin-mediated augmentation of cell spreading was inhibited not only by an antibody against human vitronectin receptor (LM 609) but also by an antibody against the beta1 subunit of integrin (mAb13), suggesting that the beta1-class integrin together with a vitronectin receptor, alphavbeta3, is mobilized onto the surface of fibroblasts upon contact with the fibrin monomer monolayer.
Asakura
S
S
Division of Hemostasis and Thrombosis Research, Jichi Medical School, Minamikawachi-Machi, Tochigi-Ken 329-04, Japan.
Niwa
K
K
Tomozawa
T
T
Jin
Y m
Ym
Madoiwa
S
S
Sakata
Y
Y
Sakai
T
T
Funayama
H
H
Soe
G
G
Forgerty
F
F
Hirata
H
H
Matsuda
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D015703:Antigens, CD / Q000378:metabolism*; D002448:Cell Adhesion; D005337:Fibrin / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism*; D039661:Integrin beta3; D051379:Mice; D009842:Oligopeptides / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000378:metabolism; D019030:Receptors, Vitronectin / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D019012:Integrin beta1; D039661:Integrin beta3; D009842:Oligopeptides; D010446:Peptide Fragments; D010980:Platelet Membrane Glycoproteins; D011971:Receptors, Immunologic; D019030:Receptors, Vitronectin; C047981:arginyl-glycyl-aspartic acid; D005337:Fibrin