Monoclonal antibody characterization of two distant sites required for function of the central cell-binding domain of fibronectin in cell adhesion, cell migration, and matrix assembly.

Nagai, Toshihiko; Yamakawa, Naomi; Aota, Shin-Ichin; Yamada, Susan S.; Akiyama, Steven K.; Olden, Kenneth; Yamada, Kenneth M.

doi:10.1083/jcb.114.6.1295

Cited by 150 publications

(94 citation statements)

References 56 publications

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“…Therefore, the relative contribution of the RGD sequence alone to the overall binding affinity strongly differs between these integrins, and the simple presence of an appropriately exposed RGD motif might be sufficient to mediate binding of ~VI]3. In contrast, recognition by integrins alIb133 and c~5131 seems to require additional binding criteria, as exemplified by the well-defined synergy region in fibronectin [29]. The absence of such additional binding structures in fibrillin-1 might thus explain an insufficient affinity for integrins ~Ilbl~3 and c~5131.…”

Section: Discussionmentioning

confidence: 99%

Cell adhesion and integrin binding to recombinant human fibrillin‐1

et al. 1996

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Fibrillin-1 is a major constituent of tissue microfibrfls that occur in most connective tissues, either in close association with or independent of elastin. To test possible cell-adhesive functions of this protein, we used recombinant human fibrlllin-1 polypeptides produced in a mammalian expression system in cell attachment and solid-phase integrin binding assays. necessitates the use of buffers containing denaturing agents and cannot separate fibrillin-1 and fibrillin-2 [18], we chose to use recombinant human fibrillin-1 polypeptides for a study of possible cell-adhesive functions. These human fibrillin-1 peptides were produced in a mammalian expression system and purified under non-denaturing conditions [19]. Extensive characterization indicated correct folding [19]. Using an experimental approach already applied to characterize cell-adhesive and integrin-binding functions of fibulins [20], laminin [21] and collagen VI [22], we demonstrate the ability of cells to adhere to fibrillin-1 via an RGD motif located in the fourth 8-cysteine domain. In addition we identify integrin aV[33 as a major cellular receptor for this site.

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Section: Discussionmentioning

confidence: 99%

Cell adhesion and integrin binding to recombinant human fibrillin‐1

et al. 1996

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“…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)(4)(5)(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).…”

Section: Fibronectin (Fn)mentioning

confidence: 99%

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: Role Of Fibronectin In Ovarian Cancermentioning

confidence: 99%

“…Although fibronectin has many biological activities, its role in cancer is well documented: promotion of cell adhesion and migration, key steps in the metastatic process (Nagai et al 1991, Kenny et al 2008. Fibronectin celladhesive sites have been identified in protease-resistant fragments of 110-120, 75 and 37 kDa, which are derived from the internal section of the protein (Ruoslahti et al 1981, Hayashi & Yamada 1983, Zardi et al 1985, Nagai et al 1991. The cell-adhesive activity attributed to these cleaved fragments is mediated via an Arg-Gly-Asp (RGD) motif that serves as a ligand recognition site for several integrins (Pierschbacher & Ruoslahti 1984, Yamada & Kennedy 1984.…”

Section: Role Of Fibronectin In Ovarian Cancermentioning

confidence: 99%

WOMEN IN CANCER THEMATIC REVIEW: Ovarian cancer–peritoneal cell interactions promote extracellular matrix processing

Ricciardelli

Lokman

Ween

et al. 2016

Endocrine-Related Cancer

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Ovarian cancer has a distinct tendency for metastasising via shedding of cancerous cells into the peritoneal cavity and implanting onto the peritoneum that lines the pelvic organs. Once ovarian cancer cells adhere to the peritoneal cells, they migrate through the peritoneal layer and invade the local organs. Alterations in the extracellular environment are critical for tumour initiation, progression and intra-peritoneal dissemination. To increase our understanding of the molecular mechanisms involved in ovarian cancer metastasis and to identify novel therapeutic targets, we recently studied the interaction of ovarian cancer and peritoneal cells using a proteomic approach. We identified several extracellular matrix (ECM) proteins including, fibronectin, TGFBI, periostin, annexin A2 and PAI-1 that were processed as a result of the ovarian cancerperitoneal cell interaction. This review focuses on the functional role of these proteins in ovarian cancer metastasis. Our findings together with published literature support the notion that ECM processing via the plasminogen-plasmin pathway promotes the colonisation and attachment of ovarian cancer cells to the peritoneum and actively contributes to the early steps of ovarian cancer metastasis.

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Monoclonal antibody characterization of two distant sites required for function of the central cell-binding domain of fibronectin in cell adhesion, cell migration, and matrix assembly.

Cited by 150 publications

References 56 publications

Cell adhesion and integrin binding to recombinant human fibrillin‐1

Cell adhesion and integrin binding to recombinant human fibrillin‐1

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

WOMEN IN CANCER THEMATIC REVIEW: Ovarian cancer–peritoneal cell interactions promote extracellular matrix processing

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