Rapid methods for isolation of human plasma fibronectin

Miekka, Shirley I.; Ingham, Kenneth C.; Ménaché, D

doi:10.1016/0049-3848(82)90272-9

Cited by 311 publications

(194 citation statements)

References 26 publications

Supporting

Mentioning

192

Contrasting

Order By: Relevance

“…Collagen I was obtained from Upstate Biotechnology (Lake Placid, NY). Human plasma fibronectin was isolated from Cohn's fraction I and II, as described previously (23). Anti-fibronectin III-1 IgG (9D2; see Ref.…”

Section: Methodsmentioning

confidence: 99%

Fibronectin matrix polymerization regulates small airway epithelial cell migration

Hocking

Chang

2003

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

The continuous conversion of soluble fibronectin into extracellular matrix fibrils occurs through a dynamic, cell-dependent process. As the extracellular matrix is assembled, changes in the conformation of matrix proteins may expose biologically active, matricryptic sites that alter cell behavior. In this study, an in vitro model of wound healing was used to determine the role of matrix fibronectin in airway epithelial cell motility. Our findings indicate that, under basal conditions, small airway epithelial cell (SAEC) migration requires active fibronectin matrix polymerization. Furthermore, SAEC migration is increased significantly by the interaction of cells with a recombinant construct containing fibronectin's matricryptic III-1 site. In contrast, addition of increasing amounts of fibronectin to SAECs significantly decreased the rate of cell migration. This fibronectin-induced inhibition of cell migration was overcome by blocking excess fibronectin matrix deposition. These data indicate that SAEC migration is regulated in a biphasic manner by the polymerization of fibronectin in the extracellular matrix and suggest a stimulatory role for fibronectin's matricryptic III-1 site in cell motility.

show abstract

Section: Methodsmentioning

confidence: 99%

Fibronectin matrix polymerization regulates small airway epithelial cell migration

Hocking

Chang

2003

American Journal of Physiology-Lung Cellular and Molecular Phys

View full text Add to dashboard Cite

show abstract

“…Chloroquine, soybean trypsin inhibitor, and bovine serum albumin were purchased from Sigma. Fibronectin was prepared from human plasma as previously described (16). GRGDSP and GRGESP peptides were obtained from Peninsula Laboratories (Belmont, CA).…”

Section: Methodsmentioning

confidence: 99%

Cell Adhesion Regulates Ubiquitin-mediated Degradation of the Platelet-derived Growth Factor Receptor β

Baron

Schwartz

2000

Journal of Biological Chemistry

View full text Add to dashboard Cite

Cross-talk between integrin-mediated adhesion and growth factors has been described in many recent studies; however, the underlying mechanisms remain incompletely understood. We report here that detachment of cells from the extracellular matrix induced a decrease in both the autophosphorylation and protein levels of the platelet-derived growth factor receptor ␤ (PDGF-R ␤), which was completely reversed upon replating cells on fibronectin. The effect occurred in all cells examined but to a greater extent in primary fibroblasts compared with established cell lines. Decreased PDGF-R levels in suspended cells correlated with ubiquitination of the PDGF-R and was blocked by treatment with inhibitors of the proteasome pathway. Unlike PDGF-induced down-regulation, detachment-induced degradation did not require receptor autophosphorylation, internalization, or tyrosine kinase activity. We conclude that cell detachment results in cellular desensitization to PDGF that is mediated by degradation of the PDGF-R via a novel ubiquitin-dependent pathway.Cell adhesion to the extracellular matrix induces a wide array of intracellular signals mediated primarily by members of the integrin family of transmembrane receptors. Integrins act coordinately with growth factors to regulate migration, growth or differentiation, and survival (reviewed in Ref. 1). For instance, several recent studies reported a synergistic effect of PDGF 1 and cell adhesion on autophosphorylation of the PDGF-R and activation of the mitogen-activated protein kinase cascade (2, 3). Moreover, PDGF induces the association of its phosphorylated receptor with integrin ␣ v ␤ 3 in fibroblasts plated on vitronectin, which correlates with enhanced PDGFinduced cell proliferation (4). These effects appear to contribute to the anchorage dependence of growth (1).PDGF-Rs are transmembrane tyrosine kinases that mediate the effects of PDGF on growth and motility in cell types including fibroblasts and smooth muscle cells (5-7). Two structurally related receptors, ␣ and ␤, possess different affinities for the A and B PDGF isoforms and generate similar though not identical cellular responses. Thus, both the ␣ and ␤ receptors stimulate cell proliferation, membrane ruffling, and loss of actin stress fibers, but only the ␤ receptor promotes chemotaxis (5, 6). Binding of PDGF to its receptor induces receptor dimerization and autophosphorylation on several tyrosine residues, which provide docking sites for SH2-containing molecules, thereby initiating a variety of signaling pathways.c-Cbl, one of the numerous proteins that bind to the PDGF-R (8), has recently been identified as a proteasomal E3-like ubiquitin ligase that is involved in down-regulation of tyrosine kinase receptors (9 -11). PDGF-induced down-regulation is a complex process that involves receptor internalization through clathrin-coated pits and requires the receptor tyrosine kinase activity (12, 13). The 100-kDa GTPase dynamin is responsible for pinching off of the vesicles, which results in the delivery of receptors to l...

show abstract

“…Human fibronectin was purified from Cohn's fractions I and 2 (a generous gift from Dr. Ken Ingham, American Red Cross, Bethesda, MD) as previously described (Miekka et al, 1982). Production and purification of recombinant rat 70-and 40-kDa fibronectin fragments and recombinant wild-type fibronectin and fibronectin lacking the RGD site (FN⌬RGD) have been previously described (Sottile and Mosher, 1997;Sottile et al, 2000).…”

Section: Proteinsmentioning

confidence: 99%

Fibronectin Polymerization Regulates the Composition and Stability of Extracellular Matrix Fibrils and Cell-Matrix Adhesions

Sottile

Hocking

2002

MBoC

552

510

View full text Add to dashboard Cite

Remodeling of extracellular matrices occurs during development, wound healing, and in a variety of pathological processes including atherosclerosis, ischemic injury, and angiogenesis. Thus, identifying factors that control the balance between matrix deposition and degradation during tissue remodeling is essential for understanding mechanisms that regulate a variety of normal and pathological processes. Using fibronectin-null cells, we found that fibronectin polymerization into the extracellular matrix is required for the deposition of collagen-I and thrombospondin-1 and that the maintenance of extracellular matrix fibronectin fibrils requires the continual polymerization of a fibronectin matrix. Further, integrin ligation alone is not sufficient to maintain extracellular matrix fibronectin in the absence of fibronectin deposition. Our data also demonstrate that the retention of thrombospondin-1 and collagen I into fibrillar structures within the extracellular matrix depends on an intact fibronectin matrix. An intact fibronectin matrix is also critical for maintaining the composition of cell-matrix adhesion sites; in the absence of fibronectin and fibronectin polymerization, neither ␣5␤1 integrin nor tensin localize to fibrillar cell-matrix adhesion sites. These data indicate that fibronectin polymerization is a critical regulator of extracellular matrix organization and stability. The ability of fibronectin polymerization to act as a switch that controls the organization and composition of the extracellular matrix and cell-matrix adhesion sites provides cells with a means of precisely controlling cell-extracellular matrix signaling events that regulate many aspects of cell behavior including cell proliferation, migration, and differentiation. INTRODUCTIONExtracellular matrix remodeling plays an important role during development, wound healing, atherosclerosis, ischemic injury, and angiogenesis. Perturbing matrix remodeling by preventing the turnover of collagen I or by altering the levels of matrix-degrading proteases or protease inhibitors has been shown to result in fibrosis, arthritis, reduced angiogenesis, and developmental abnormalities (Liu et al., 1995;Vu et al., 1998;Holmbeck et al., 1999;Ducharme et al., 2000). In normal adult tissue, some extracellular matrix components such as elastin and fibrillar collagen have half lives of months to years (Krane, 1985;Debelle and Tamburro, 1999). Other extracellular matrix components, such as proteoglycans, thrombospondin-1 and -2, and vitronectin, can be endocytosed and degraded in the lysosomes (McKeownLongo et al., 1984;Yanagishita and Hascall, 1984; MurphyUllrich and Mosher, 1987;Hausser et al., 1992;Godyna et al., 1995a;Pijuan-Thompson and Gladson, 1997;Memmo and McKeown-Longo, 1998). Extracellular matrix molecules can also be degraded extracellularly by proteases such as matrix metalloproteinases (MMPs), plasminogen activators, and plasmin (Hynes, 1990;Marchina and Barlati, 1996;Shapiro, 1998).Recent data indicate that polymerized forms of extracellular matri...

show abstract

Rapid methods for isolation of human plasma fibronectin

Cited by 311 publications

References 26 publications

Fibronectin matrix polymerization regulates small airway epithelial cell migration

Fibronectin matrix polymerization regulates small airway epithelial cell migration

Cell Adhesion Regulates Ubiquitin-mediated Degradation of the Platelet-derived Growth Factor Receptor β

Fibronectin Polymerization Regulates the Composition and Stability of Extracellular Matrix Fibrils and Cell-Matrix Adhesions

Contact Info

Product

Resources

About