Role of the I-9 and III-1 modules of fibronectin in formation of an extracellular fibronectin matrix.
Cultured fibroblasts bind soluble protomeric fibronectin and mediate its conversion to insoluble disulfide-bonded multimers. The disulfide-bonded multimers are deposited in fibrillar pericellular matrix. Antifibronectin monoclonal antibodies were analyzed to identify domains of fibronectin required for assembly into matrix. Two antibodies, L8 and 9D2, inhibited binding and insolubilization of 125I-labeled plasma fibronectin by fibroblasts but did not inhibit binding of labeled amino-terminal 70-kDa fragment of fibronectin to matrix assembly sites. Immunoblotting of fibronectin fragments showed that the epitope for 9D2 is in the first type III homology sequence (III-1) whereas the epitope for L8 requires that the last type I sequence of the gelatin binding region (I-9) be contiguous to III-1 and is sensitive to reduction of disulfides in I-9. A 56-kDa gelatin-binding thermolysin fragment of fibronectin that contains III-1 and the L8 and 9D2 epitopes inhibited binding of fibronectin to cell layers 10-fold better than a 40-kDa gelatin-binding fragment that lacks III-1 and the antigenic sites. This 56-kDa fragment, however, did not bind specifically to cell layers. These results indicate that the I-9 and III-1 modules of fibronectin form a functional unit that mediates an interaction, perhaps between protomers, important in the assembly of fibronectin.
Chernousov
M A
MA
Institute of Experimental Cardiology, USSR Cardiology Research Center, Academy of Medical Sciences, Moscow.
Fogerty
F J
FJ
Koteliansky
V E
VE
Mosher
D F
DF
D000911:Antibodies, Monoclonal; D001665:Binding Sites; D001666:Binding Sites, Antibody; D002478:Cells, Cultured; D000939:Epitopes / Q000032:analysis; D005109:Extracellular Matrix / Q000502:physiology*; D005347:Fibroblasts / Q000502:physiology; D005353:Fibronectins / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D007231:Infant, Newborn; D007700:Kinetics; D008297:Male; D008961:Models, Structural; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D012879:Skin Physiological Phenomena
D000911:Antibodies, Monoclonal; D000939:Epitopes; D005353:Fibronectins; D010446:Peptide Fragments