Assembly of fibronectin fibrils occurs at the surface of substrate-attached cells and is mediated by the first to the fifth type I modules in the N-terminal 70 kDa portion of the molecule. The first type III module (III1) of fibronectin, not present in the 70 kDa portion, contains a conformation-dependent binding site for the 70 kDa N-terminal region of fibronectin, suggesting that the III1 module on cell-surface fibronectin may serve as a binding site for fibronectin's N-terminus on substrate-attached cells. To explore this possiblility, we compared the ability of mutant recombinant 70 kDa proteins containing deletions of one or several of the first five type I modules to bind to fibroblasts and to III1. Proteins containing the fourth and fifth type I modules (70KΔI1-3) bound specifically to III1 in solid-phase binding assays; proteins lacking I4 and I5 did not bind. N-terminal molecules containing the fourth and fifth type I modules also bound to fibroblasts, suggesting that III1-like binding sites are present on the cell surface. However, the high-affinity binding sites on fibroblasts for fibronectin or the 70 kDa protein displayed more complex determinants, inasmuch as 70 kDa deletion mutants lacking I4 and I5 also bound to the cell surface, and deletion mutants lacking I1-3 and I4-5 both competed only partially for binding of 125I-labelled fibronectin or 70 kDa protein. These data indicate that the N-terminal part of fibronectin binds to III1 via I4 and I5 and that interactions in addition to that of I4 and I5 with III1 are important for cell-surface-mediated fibronectin polymerization.
Research Article|April 01 1997
N-terminal type I modules required for fibronectin binding to fibroblasts and to fibronectin's III1 module
*Department of Physiology and Cell Biology (A-134), Neil Hellman Medical Research Building, Albany Medical College of Union University, 47 New Scotland Avenue, Albany, NY 12208
‡To whom correspondence should be sent.
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Jane SOTTILE, Deane F. MOSHER; N-terminal type I modules required for fibronectin binding to fibroblasts and to fibronectin's III1 module. Biochem J 1 April 1997; 323 (1): 51–60. doi: https://doi.org/10.1042/bj3230051
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