Monoclonal antibody to fibronectin which inhibits extracellular matrix assembly

Chernousov, Michael A.; Faerman, Alexander; Frid, Maria G.; Printseva, O. Yu.; Koteliansky, Victor

doi:10.1016/0014-5793(87)81255-3

Cited by 34 publications

(23 citation statements)

References 15 publications

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“…One possible explanation is that domain III-1 is opened in fibrils, but the cysteine becomes buried in an FN-FN bond. Previous work has implicated III-1 in FN fibril assembly (27)(28)(29). Conversely, domains III-6 and III-12 showed no labeling in solution but showed significant labeling in stretched fibrils, suggesting that these domains are unfolded specifically in matrix fibrils.…”

Section: Table 2 Relative Rates Of Fn-iii Domain Openingmentioning

confidence: 83%

Probing the Folded State of Fibronectin Type III Domains in Stretched Fibrils by Measuring Buried Cysteine Accessibility

Lemmon

Ohashi

Erickson

2011

Journal of Biological Chemistry

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Fibronectin (FN) is an extracellular matrix protein that is assembled into fibrils by cells during tissue morphogenesis and wound healing. FN matrix fibrils are highly elastic, but the mechanism of elasticity has been debated: it may be achieved by mechanical unfolding of FN-III domains or by a conformational change of the molecule without domain unfolding. Here, we investigate the folded state of FN-III domains in FN fibrils by measuring the accessibility of buried cysteines. Four of the 15 FN-III domains (III-2, -3, -9, and -11) appear to unfold in both stretched fibrils and in solution, suggesting that these domains spontaneously open and close even in the absence of tension. Two FN-III domains (III-6 and -12) appear to unfold only in fibrils and not in solution. These results suggest that domain unfolding can at best contribute partially to the 4-fold extensibility of fibronectin fibrils. Fibronectin (FN)2 is a dimer of a 250-kDa protein that forms insoluble fibrils that are a critical component of the extracellular matrix. It is made up of a series of three domain types connected end-to-end (see Fig. 1A) (1). The N terminus of FN consists of nine FN-I and two FN-II domains, each of which contains an internal disulfide bond. The C terminus of FN consists of three FN-I domains and contains intermolecular disulfides that link the two FN monomers to form the dimer. The central portion of the FN molecule is made up of 15 tandem FN-III domains; these domains each consist of seven ␤-strands folded into a ␤-sandwich structure and contain no internal disulfides (see Fig. 1B). FN in solution adopts a compact conformation in which the second to fourth FN-III domains (III-2-4) of one subunit interact with the 12 th to 14 th FN-III domains (III-12-14) of the other, causing the FN dimer to fold onto itself (see Fig. 1A) (2).FN interacts with both cells and other extracellular matrix proteins. The N-terminal domains contain binding sites for the extracellular matrix proteins collagen and fibrin, whereas the ninth and tenth FN-III domains bind to integrins at the cell surface (3-6). Upon attachment to cells, FN is stretched and assembled into fibrils by cell contraction: matrix assembly is inhibited if cell contractility is inhibited (7) or if tension within the matrix is released (8). Although the exact mechanism of FN fibril formation is still unknown, studies have shown that FN fibrils are stretched to up to 4 times their resting length by cells and are highly elastic structures (9 -12). This elasticity requires cell contraction; disruption of cytoskeletal contraction relaxes stretched fibrils (10).Two different mechanisms have been proposed to explain the elasticity of FN matrix fibrils (see Fig. 1A) (13). One theory contends that the elasticity is a function of FN-III domains unfolding under tension (see Fig. 1A, mechanism 2) (14, 15). These domains contain no internal disulfides and can be mechanically unfolded by atomic force microscopy (16, 17). It is unclear how relevant these atomic force microscopy pulling force...

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Section: Table 2 Relative Rates Of Fn-iii Domain Openingmentioning

confidence: 83%

Probing the Folded State of Fibronectin Type III Domains in Stretched Fibrils by Measuring Buried Cysteine Accessibility

Lemmon

Ohashi

Erickson

2011

Journal of Biological Chemistry

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“…The first FNIII domain consists of a beta sandwich that may be stretched into an open conformation by local cell traction forces. A cryptic site is revealed in this open configuration that is important for intermolecular fibronectin binding, and this site contains a specific epitope for the L8 monoclonal antibody (Chernousov et al, 1987;Hocking et al, 1994;Smith et al, 2007;Zhong et al, 1998). The L8 antibody labeled Fig.…”

Section: Discussionmentioning

confidence: 96%

Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix

2009

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“…Remodeling of the fibronectin matrix in response to III1c occurs rapidly, does not require the presence of cells and is not accompanied by changes in the levels of fibronectin matrix. Previous studies have indicated that the III1 region of fibronectin participates in homophilic binding interactions among fibronectin molecules, which can regulate the assembly of the fibronectin matrix (Chernousov et al, 1987;Chernousov et al, 1991;Hocking et al, 1994;Hocking et al, 1996;Morla et al, 1994;Morla and Ruoslahti, 1992). Reports on the effects of the III1c peptide on the IST-9 Fig.…”

Section: Discussionmentioning

confidence: 99%

Stimulation of extracellular matrix remodeling by the first type III repeat in fibronectin

Klein

Zheng

Ambesi

et al. 2003

Journal of Cell Science

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The fibronectin matrix contains cryptic sites which are thought to modulate cellular biological responses. One of these sites, located in fibronectin's first type III repeat (III1c), influences signaling pathways that are relevant to cytoskeletal organization and cell cycle progression. The purpose of this study was to identify possible mechanisms responsible for the effects of III1c on cell behavior. Recombinant peptides representing various type III repeats of fibronectin were compared for their effects on fibronectin matrix organization and activation of intracellular signaling pathways. III1c and III13 but not III11c or III10 bound to monolayers of human skin fibroblasts in a dose- and time-dependent manner and were localized to the extracellular matrix. Binding of III13, but not III1c, to matrix was sensitive to heparitinase, suggesting that the association of III1c with the matrix was not dependent on heparan sulfate proteoglycans. Quantitative and morphological assessment indicated that, in contrast to previously published reports, the binding of III1c to cell layers did not result in the loss or disruption of matrix fibronectin. Binding of III1c but not III13 to the extracellular matrix did result in the loss of a conformationally sensitive epitope present within the EDA type III module of cellular fibronectin. III1c-induced loss of the EDA epitope did not require the presence of cells, occurred within 1 hour and was associated with the activation of p38 mitogen-activated protein kinase (MAPK) followed by the formation of filopodia. Maximal phosphorylation of p38 MAPK occurred within 1 hour, whereas cytoskeletal changes did not appear until 12 hours later. These findings are consistent with a model in which the binding of III1c to the extracellular matrix results in a conformational remodeling of the fibronectin matrix, which has both short- and long-term effects on cell physiology.

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Monoclonal antibody to fibronectin which inhibits extracellular matrix assembly

Cited by 34 publications

References 15 publications

Probing the Folded State of Fibronectin Type III Domains in Stretched Fibrils by Measuring Buried Cysteine Accessibility

Probing the Folded State of Fibronectin Type III Domains in Stretched Fibrils by Measuring Buried Cysteine Accessibility

Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix

Stimulation of extracellular matrix remodeling by the first type III repeat in fibronectin

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