Type I collagen contains at least 14 cryptic fibronectin binding sites of similar affinity

Ingham, Kenneth C.; Brew, Shelesa A.; Migliorini, Mary

doi:10.1016/s0003-9861(02)00454-x

Cited by 29 publications

(62 citation statements)

References 19 publications

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“…To explore the FN-collagen interaction, we tested by NMR a series of synthetic single-stranded peptides covering ␣ 1 (I) residues G 763 -R 816 for binding the 2 subfragments of the GBD, [8][9] FnI and 6 FnI 1-2 FnII 7 FnI. The tested region of collagen spans the MMP-1 cleavage site, G 775 /I 776 , which has been shown to influence FN-collagen interactions (6).…”

Section: Gbd Interacts With a Type I Collagen Peptide Adjacent To Thementioning

confidence: 99%

“…Peptides spanning that region (15) or MMP-1 cleavage at this site (6) inhibit attachment. However, attempts to reconstitute the FN-collagen interaction in vitro by using synthetic peptides failed to find strong, specific, interactions (8,9), and the precise sequence determinants for FN-binding to collagen remained unknown.Here, we report a tight interaction between GBD and a synthetic peptide from the ␣ 1 chain of collagen type I [␣ 1 (I)], defined by using NMR and fluorescence. Both GBD subfragments bind the same peptide sequence just C-terminal to the MMP-1 cleavage site; the tightest binding, however, is mediated by the [8][9] FnI domain pair that also binds ␣ 2 (I).…”

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confidence: 99%

“…The tested region of collagen spans the MMP-1 cleavage site, G 775 /I 776 , which has been shown to influence FN-collagen interactions (6). Of the FN fragments, [8][9] FnI is known to associate with ␣ 1 (I) fragments and gelatin (16,17), and 6 FnI 1-2 FnII 7 FnI binds gelatin more strongly than the shorter 6 FnI 1-2 FnII (18). The NMR titrations in this study often resulted in unfavorable intermediate exchange regimes; therefore, we tried to optimize the experimental conditions in each case by using different temperatures and different magnetic field strengths, 500 and 950 MHz.…”

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confidence: 99%

“…Both GBD subfragments only bound tightly to a peptide spanning ␣ 1 (I) residues G 778 -G 799 (Fig. 2 A and B), with dissociation constants, K d , of 5 Ϯ 1 M for [8][9] FnI (25°C) and 57 Ϯ 6 M for …”

mentioning

confidence: 99%

“…Both GBD subfragments bind the same peptide sequence just C-terminal to the MMP-1 cleavage site; the tightest binding, however, is mediated by the [8][9] FnI domain pair that also binds ␣ 2 (I). A 2.1-Å crystal structure of [8][9] FnI in complex with the ␣ 1 (I) peptide shows an extended single-strand peptide conformation, which suggests that FN stabilizes a noncanonical collagen structure upon binding. We explore this hypothesis in thermal denaturation experiments using model triple-helical peptides.…”

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confidence: 99%

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Identification and structural analysis of type I collagen sites in complex with fibronectin fragments

Erat¹,

Slatter

Lowe

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

136

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Collagen and fibronectin are major components of vertebrate extracellular matrices. Their association and distribution control the development and properties of diverse tissues, but thus far no structural information has been available for the complex formed. Here, we report binding of a peptide, derived from the ␣1 chain of type I collagen, to the gelatin-binding domain of human fibronectin and present the crystal structure of this peptide in complex with the 8 -9 FnI domain pair. Both gelatin-binding domain subfragments, 6 FnI 1-2 FnII 7 FnI and 8 -9 FnI, bind the same specific sequence on D-period 4 of collagen I ␣1, adjacent to the MMP-1 cleavage site. 8 -9 FnI also binds the equivalent sequence of the ␣2 chain. The collagen peptide adopts an antiparallel ␤-strand conformation, similar to structures of proteins from pathogenic bacteria bound to FnI domains. Analysis of the type I collagen sequence suggests multiple putative fibronectin-binding sites compatible with our structural model. We demonstrate, by kinetic unfolding experiments, that the triple-helical collagen state is destabilized by 8 -9 FnI. This finding suggests a role for fibronectin in collagen proteolysis and tissue remodeling.collagen destabilization ͉ extracellular matrix ͉ protein structure C ollagen is the most abundant protein in mammals, accounting for Ϸ25% of the body-protein content. It is crucial for effects as diverse as cell differentiation, cell migration, and mechanical properties of tissues. Many human diseases have their origin in mutations that affect interactions of collagen with other extracellular matrix (ECM) molecules and cell receptors (1). Type I collagen fibrils form spontaneously in vitro; in vivo, however, formation requires integrin receptors and fibronectin (FN) (2). FN is a large glycosylated protein composed of multiple copies of 3 classes of domains, FnI, FnII, and FnIII, that forms fibrils in the ECM (3). It plays a role in many important physiological processes, such as embryogenesis, wound healing, hemostasis, and thrombosis (4), and disruption of the FN gene is embryonic lethal in mice (5).The interaction of these 2 proteins has long been demonstrated in vitro by using denatured collagen (gelatin) (6, 7) and isolated collagen type I chains or chain fragments (8, 9). The collagen-binding site on FN has been localized to the 42-kDa gelatin-binding domain (GBD; for an overview of FN and collagen fragments see Fig. 1) (10, 11), and anti-GBD antibodies inhibit collagen organization in fibroblast cultures (12). Similarly, experiments in cultures have identified the collagenase/ MMP-1 (13) cleavage site in type I collagen as important for FN-mediated attachment of fibroblasts to collagen ECM (14). Peptides spanning that region (15) or MMP-1 cleavage at this site (6) inhibit attachment. However, attempts to reconstitute the FN-collagen interaction in vitro by using synthetic peptides failed to find strong, specific, interactions (8,9), and the precise sequence determinants for FN-binding to collagen remained unknown...

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Section: Gbd Interacts With a Type I Collagen Peptide Adjacent To Thementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Identification and structural analysis of type I collagen sites in complex with fibronectin fragments

Erat¹,

Slatter

Lowe

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

136

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Cell spreading and proliferation in response to the composition and mechanics of engineered fibrillar extracellular matrices

Chen

DelRio

Peterson

et al. 2013

Biotech & Bioengineering

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The extracellular matrix (ECM) consists of a complex mixture of biochemical and physical stimuli that together regulate cell behavior. In this study, we engineer a model ECM consisting of fibrillar Type-1 collagen plus fibronectin that allows systematic examination of the effects of matrix composition and mechanics on cells. On this combined protein matrix, cells exhibit intermediate degrees of spreading and proliferation compared to their responses on collagen or fibronectin alone. Adhesion to the combination matrix could be blocked by peptides containing the sequence arginine-glycine-aspartic acid (RGD) and by antibodies against α1 integrin, suggesting cell-matrix engagement was mediated by a combination of integrin receptors that recognize fibronectin and collagen. Regardless of integrin engagement, cells were sensitive to the mechanical properties of the combination ECM, suggesting that cells could process biochemical and mechanical cues simultaneously and independently.

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Matrix‐mediated retention of in vitro osteogenic differentiation potential and in vivo bone‐forming capacity by human adult bone marrow‐derived mesenchymal stem cells during ex vivo expansion

Mauney

Kirker‐Head

Abrahamson

et al. 2006

J Biomedical Materials Res

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Mesenchymal stem cells (MSCs) represent an attractive cell source for tissue engineering applications, since they are readily isolated from adult bone marrow and have the ability to differentiate along multiple mesenchymal lineages, including osteogenic. Currently, utilization of MSCs for bone tissue engineering is limited because of the attenuation of their osteogenic differentiation potential and in vivo bone-forming capacity following ex vivo expansion on conventional tissue culture plastic (TCP). Previously, we demonstrated that a denatured type I collagen (DC) matrix promotes the maintenance of MSC in vitro osteogenic differentiation potential during ex vivo expansion in contrast to TCP. In this study, we further demonstrate that the maintenance of MSC osteogenic differentiation potential is primarily due to the ability of DC matrix to influence the retention of early passage osteogenic functions in late passage (LP) cells during ex vivo expansion, in contrast to solely enhancing attenuated LP cellular functions during osteogenic differentiation. Serum-associated factors played a significant role in influencing the retention of MSC osteogenic differentiation potential during expansion on the DC matrix. Significantly, the results show that although LP cells expanded ex vivo on TCP highly attentuate their in vivo bone-forming capacity, the expansion of MSCs on DC matrix preserves this ability as determined by histological, histomorphometric, and bone mineral density evaluations of MSC-seeded hydroxyapatite/tricalcium phosphate scaffolds following an 8-week implantation period within a heterotopic muscle pouch model. These findings provide further insight into the importance of matrix-mediated effects on MSC function and selective factors important in this process.

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Type I collagen contains at least 14 cryptic fibronectin binding sites of similar affinity

Cited by 29 publications

References 19 publications

Identification and structural analysis of type I collagen sites in complex with fibronectin fragments

Identification and structural analysis of type I collagen sites in complex with fibronectin fragments

Cell spreading and proliferation in response to the composition and mechanics of engineered fibrillar extracellular matrices

Matrix‐mediated retention of in vitro osteogenic differentiation potential and in vivo bone‐forming capacity by human adult bone marrow‐derived mesenchymal stem cells during ex vivo expansion

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