scholarly journals A novel fibronectin binding site required for fibronectin fibril growth during matrix assembly

2001 ◽  
Vol 154 (5) ◽  
pp. 1081-1088 ◽  
Author(s):  
Jan L. Sechler ◽  
Hongwei Rao ◽  
Anne Marie Cumiskey ◽  
Irbert Vega-Colón ◽  
Michael S. Smith ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Binding Site ◽  
Large Deletion ◽  
Specific Function ◽  
Binding Assays ◽  
Cell Binding ◽  
Matrix Assembly ◽  
Type Iii ◽  
Fibronectin Binding ◽  
Cell Binding Domain

Fibronectin (FN) assembly into a fibrillar extracellular matrix is a stepwise process requiring participation from multiple FN domains. Fibril formation is regulated in part by segments within the first seven type III repeats (III1–7). To define the specific function(s) of this region, recombinant FNs (recFNs) containing an overlapping set of deletions were tested for the ability to assemble into fibrils. Surprisingly, recFN lacking type III repeat III1 (FNΔIII1), which contains a cryptic FN binding site and has been suggested to be essential for fibril assembly, formed a matrix identical in all respects to a native FN matrix. Similarly, displacement of the cell binding domain in repeats III9–10 to a position close to the NH2-terminal assembly domain, as well as a large deletion spanning repeats III4–7, had no effect on assembly. In contrast, two deletions that included repeat III2, ΔIII1–2 and ΔIII2–5, caused significant reductions in fibril elongation, although binding of FN to the cell surface and initiation of assembly still proceeded. Using individual repeats in binding assays, we show that III2 but not III1 contains an FN binding site. Thus, these results pinpoint repeat III2 as an important module for FN–FN interactions during fibril growth.

scholarly journals A fibronectin self-assembly site involved in fibronectin matrix assembly: reconstruction in a synthetic peptide.

1992 ◽  
Vol 118 (2) ◽  
pp. 421-429 ◽  
Author(s):  
A Morla ◽  
E Ruoslahti
Keyword(s):  
Extracellular Matrix ◽  
Binding Site ◽  
Self Assembly ◽  
Synthetic Peptide ◽  
Active Form ◽  
Matrix Form ◽  
Matrix Assembly ◽  
Type Iii ◽  
Fibronectin Matrix ◽  
Fibronectin Binding

The active form of fibronectin is its extracellular matrix form, which allows for the attachment of cells and influences both the growth and migration of cells. The matrix form is assembled by cells; however, many cells are defective in this regard. Several regions within fibronectin have been shown to play a role in matrix assembly by cells. One such region has been localized into the first type III repeat of fibronectin (Chernousov, M. A., F. J. Fogerty, V. E. Koteliansky, and D. F. Mosher. J. Biol. Chem. 266:10851-10858). We have identified this site as a fibronectin-fibronectin binding site and reproduced it as a synthetic peptide. This site is contained in a 14-kD fragment that corresponds to portions of the first two type III repeats. The 14-kD fragment was found to bind to cell monolayers and to inhibit fibronectin matrix assembly. The 14-kD fragment only slightly reduced the binding of fibronectin to cell surfaces but it significantly inhibited the subsequent incorporation of fibronectin into the extracellular matrix. The 14-kD fragment also bound to purified fibronectin and inhibited fibronectin-fibronectin binding. A synthetic 31-amino acid peptide (P1) representing a segment of the 14-kD fragment retained the ability to inhibit fibronectin-fibronectin binding. Peptide P1 specifically bound fibronectin from plasma in affinity chromatography, whereas a column containing another peptide from the 14-kD fragment did not. These results define a fibronectin-fibronectin binding site that appears to promote matrix assembly by allowing the assembly of fibronectin molecules into nascent fibrils. The 14-kD fragment and the P1 peptide that contain this site inhibit matrix assembly by competing for the fibronectin-fibronectin binding.

scholarly journals Binding to Human Extracellular Matrix byNeisseria meningitidis

1998 ◽  
Vol 66 (4) ◽  
pp. 1791-1794 ◽  
Author(s):  
Thomas Eberhard ◽  
Ritva Virkola ◽  
Timo Korhonen ◽  
Göran Kronvall ◽  
Måns Ullberg
Keyword(s):  
Extracellular Matrix ◽  
Neisseria Meningitidis ◽  
Binding Site ◽  
Binding Domain ◽  
Cell Binding ◽  
Matrix Components ◽  
Cell Binding Domain ◽  
Healthy Carriers ◽  
Better Than

ABSTRACT Adhesion of Neisseria meningitidis strains to extracellular matrix (ECM) and purified matrix components was examined. Most strains bound to subendothelial ECM as well as to immobilized fibronectin and types I, III, and V collagen. Strains from healthy carriers adhered significantly better than isolates from patients. The binding site was localized to the central 75-kDa cell-binding domain of the fibronectin molecule. This domain has not been described previously to interact with bacterial structures.

scholarly journals Fibronectin and Its Role in Human Infective Diseases

Cells ◽  
2019 ◽  
Vol 8 (12) ◽  
pp. 1516 ◽  
Author(s):  
Pietro Speziale ◽  
Carla Renata Arciola ◽  
Giampiero Pietrocola
Keyword(s):  
Binding Site ◽  
Host Cells ◽  
Cell Binding ◽  
Receptor Binding Site ◽  
Number Of Microorganisms ◽  
Fibronectin Binding ◽  
Fibronectin Domains ◽  
Host Infection ◽  
Antigenic Profile ◽  
Cell Binding Domain

Fibronectin is a multidomain glycoprotein ubiquitously detected in extracellular fluids and matrices of a variety of animal and human tissues where it functions as a key link between matrices and cells. Fibronectin has also emerged as the target for a large number of microorganisms, particularly bacteria. There are clear indications that the binding of microorganism’ receptors to fibronectin promotes attachment to and infection of host cells. Each bacterium may use different receptors which recognize specific fibronectin domains, mostly the N-terminal domain and the central cell-binding domain. In many cases, fibronectin receptors have actions over and above that of simple adhesion: In fact, adhesion is often the prerequisite for invasion and internalization of microorganisms in the cells of colonized tissues. This review updates the current understanding of fibronectin receptors of several microorganisms with emphasis on their biochemical and structural properties and the role they can play in the onset and progression of host infection diseases. Furthermore, we describe the antigenic profile and discuss the possibility of designing adhesion inhibitors based on the structure of the fibronectin-binding site in the receptor or the receptor-binding site in fibronectin.

scholarly journals Role of type III homology repeats in cell adhesive function within the cell-binding domain of fibronectin.

1991 ◽  
Vol 266 (5) ◽  
pp. 3045-3051
Author(s):  
F Kimizuka ◽  
Y Ohdate ◽  
Y Kawase ◽  
T Shimojo ◽  
Y Taguchi ◽  
...  
Keyword(s):  
Binding Domain ◽  
Cell Binding ◽  
Type Iii ◽  
Cell Adhesive ◽  
Cell Binding Domain

Role of Ninth Type-III Domain of Fibronectin in the Mediation of Cell-Binding Domain Adsorption on Surfaces with Different Chemistries

Langmuir ◽  
2018 ◽  
Vol 34 (33) ◽  
pp. 9847-9855 ◽  
Author(s):  
Tianjie Li ◽  
Lijing Hao ◽  
Jiangyu Li ◽  
Chang Du ◽  
Yingjun Wang
Keyword(s):  
Binding Domain ◽  
Cell Binding ◽  
Type Iii ◽  
Cell Binding Domain

scholarly journals Induction of fibronectin matrix assembly in human fibrosarcoma cells by dexamethasone.

1987 ◽  
Vol 104 (3) ◽  
pp. 601-610 ◽  
Author(s):  
P J McKeown-Longo ◽  
C A Etzler
Keyword(s):  
Extracellular Matrix ◽  
Surface Protein ◽  
Order Rate Constant ◽  
Binding Activity ◽  
Matrix Assembly ◽  
Cultured Fibroblasts ◽  
Fibrosarcoma Cells ◽  
The Matrix ◽  
Fibronectin Binding ◽  
Human Fibrosarcoma Cells

Previous studies have suggested that the assembly of fibronectin into the extracellular matrix of cultured fibroblasts is mediated by specific matrix assembly receptors that recognize a binding site in the amino terminus of the fibronectin molecule (McKeown-Longo, P.J., and D.F. Mosher, 1985, J. Cell Biol., 100:364-374). In the presence of dexamethasone, human fibrosarcoma cells (HT-1080) acquired the ability to specifically bind exogenous plasma fibronectin and incorporate it into a detergent-insoluble extracellular matrix. Dexamethasone-induced fibronectin binding to HT-1080 cells was time dependent, dose dependent, and inhibited by cycloheximide. Saturation binding curves indicated that dexamethasone induced the appearance of 7.7 X 10(4) matrix assembly receptors per cell. The induced receptors exhibited a dissociation constant (KD) for soluble fibronectin of 5.0 X 10(-8) M. In parallel experiments, normal fibroblasts exhibited 4.1 X 10(5) receptors (KD = 5.3 X 10(-8) M) per cell. In the presence of cycloheximide, the induced fibronectin-binding activity on HT-1080 cells returned to uninduced levels within 12 h. In contrast, fibronectin-binding activity on normal fibroblasts was stable in the presence of cycloheximide for up to 54 h. The first-order rate constant (Kt = 2.07 X 10(-4) min-1) for the transfer of receptor-bound fibronectin to extracellular matrix was four- to fivefold less than that for normal fibroblasts (Kt = 1.32 X 10(-3) min-1). Lactoperoxidase-catalyzed iodination of HT-1080 monolayers indicated that a 48,000-mol-wt cell surface protein was enhanced with dexamethasone. The results from these experiments suggest that dexamethasone induces functional matrix assembly receptors on the surface of HT-1080 cells; however, the rate of incorporation of fibronectin into the matrix is much slower than that of normal fibroblasts.

scholarly journals TSG-6 binds via its CUB_C domain to the cell-binding domain of fibronectin and increases fibronectin matrix assembly

2008 ◽  
Vol 27 (3) ◽  
pp. 201-210 ◽  
Author(s):  
S KUZNETSOVA ◽  
D MAHONEY ◽  
G MARTINMANSO ◽  
T ALI ◽  
H NENTWICH ◽  
...  
Keyword(s):  
Binding Domain ◽  
Cell Binding ◽  
Matrix Assembly ◽  
Fibronectin Matrix ◽  
Cell Binding Domain

scholarly journals 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.

1991 ◽  
Vol 114 (6) ◽  
pp. 1295-1305 ◽  
Author(s):  
T Nagai ◽  
N Yamakawa ◽  
S Aota ◽  
S S Yamada ◽  
S K Akiyama ◽  
...  
Keyword(s):  
Cell Spreading ◽  
Binding Domain ◽  
Central Cell ◽  
Site Directed Mutagenesis ◽  
Cell Binding ◽  
Matrix Assembly ◽  
Fibronectin Receptor ◽  
Molar Basis ◽  
Rgd Site ◽  
Cell Binding Domain

Site-directed mutagenesis studies have suggested that additional peptide information in the central cell-binding domain of fibronectin besides the minimal Arg-Gly-Asp (RGD) sequence is required for its full adhesive activity. The nature of this second, synergistic site was analyzed further by protein chemical and immunological approaches using biological assays for adhesion, migration, and matrix assembly. Fragments derived from the cell-binding domain were coupled covalently to plates, and their specific molar activities in mediating BHK cell spreading were compared with that of intact fibronectin. A 37-kD fragment purified from chymotryptic digests of human plasma fibronectin had essentially the same specific molar activity as intact fibronectin. In contrast, other fragments such as an 11.5-kD fragment lacking NH2-terminal sequences of the 37-kD fragment had only poor spreading activity on a molar basis. Furthermore, in competitive inhibition assays of fibronectin-mediated cell spreading, the 37-kD fragment was approximately 325-fold more active than the GRGDS synthetic peptide on a molar basis. mAbs were produced using the 37-kD protein as an immunogen and their epitopes were characterized. Two separate mAbs, one binding close to the RGD site and the other to a site approximately 15 kD distant from the RGD site, individually inhibited BHK cell spreading on fibronectin by greater than 90%. In contrast, an antibody that bound between these two sites had minimal inhibitory activity. The antibodies found to be inhibitory in cell spreading assays for BHK cells also inhibited both fibronectin-mediated cell spreading and migration of human HT-1080 cells, functions which were also dependent on function of the alpha 5 beta 1 integrin (fibronectin receptor). Assembly of endogenously synthesized fibronectin into an extracellular matrix was not significantly inhibited by most of the anti-37-kD mAbs, but was strongly inhibited only by the antibodies binding close to the RGD site or the putative synergy site. These results indicate that a second site distant from the RGD site on fibronectin is crucial for its full biological activity in diverse functions dependent on the alpha 5 beta 1 fibronectin receptor. This site is mapped by mAbs closer to the RGD site than previously expected.

scholarly journals Fibronectin mediates Treponema pallidum cytadherence through recognition of fibronectin cell-binding domain.

1985 ◽  
Vol 161 (3) ◽  
pp. 514-525 ◽  
Author(s):  
D D Thomas ◽  
J B Baseman ◽  
J F Alderete
Keyword(s):  
Treponema Pallidum ◽  
Binding Domain ◽  
Host Cells ◽  
Scatchard Analysis ◽  
Cell Surfaces ◽  
Cell Binding ◽  
High Affinity ◽  
Fibronectin Binding ◽  
Affinity Interaction ◽  
Cell Binding Domain

The specificity of the interaction between Treponema pallidum and fibronectin was demonstrated. Treatment of host cells with only antifibronectin sera and not anticollagen or antilaminin sera, inhibited treponemal cytadsorption. Incubation of fibronectin-coated coverslips with monoclonal antibody to the cell-binding domain of fibronectin reduced treponemal attachment to the same extent as antifibronectin serum. Both iodinated fibronectin and iodinated cell-binding domain bound to T. pallidum in a saturable manner. Specificity of the T. pallidum association with the cell-binding domain was the most effective inhibitor of the binding of either radioiodinated cell-binding domain or fibronectin to T. pallidum. Scatchard analysis gave Kd on the order of 10(-7) M for both cell-binding domain and fibronectin binding to T. pallidum, consistent with the high affinity interaction of these organisms with host cell surfaces. Finally, the same level of attachment of treponemes was achieved on coverslips coated with cell-binding domain as that observed for organisms incubated with fibronectin, indicating that the cell-binding domain polypeptide is functionally identical to fibronectin in mediating T. pallidum adherence.

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