Glycosaminoglycans that bind cold-insoluble globulin in cell-substratum adhesion sites of murine fibroblasts.

Laterra, John; Ansbacher, Riva; Culp, Lloyd A.

doi:10.1073/pnas.77.11.6662

Cited by 102 publications

(57 citation statements)

References 26 publications

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“…have shown that heparin or heparan sulfate binds to laminin (45) and fibronectin (46)(47)(48) and stabilizes the interaction of collagen and fibronectin (49). This interaction is very specific irn that heparin or heparan sulfate with a low or high sulfate content each has a markedly different affinity for fibronectin (49).…”

Section: Resultsmentioning

confidence: 99%

Influence of collagen substrata on glycosaminoglycan production by B16 melanoma cells.

Luikart

Maniglia

Sartorelli

1983

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

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A cloned metastatic murine melanoma cell line exhibited similar growth characteristics when propagated on either type I collagen, type IV collagen, or plastic. However, cells grown on both types of collagen exhibited an altered cellular morphology and on type IV collagen only, an increased substrate adhesiveness, relative to those maintained on a plastic substratum. Incorporation of [3H]glucosamine and [5S]sulfate into glycosaminoglycans (GAGs) of cells grown on collagen substrates was 20% and 40% less, respectively, than cells grown on plastic, whereas degradation of cell-associated [35S]sulfate-labeled GAGs was similar in cells grown on collagen or plastic. Although the composition of GAGs was similar in all cultures, consisting of approximately 60% chondroitin and 40% heparin or heparan sulfate, the degree of sulfation of the heparin or heparan sulfate molecules was markedly decreased in cultures grown on collagen. The results indicate that the composition of the extracellular matrix influences the biological behavior of B16 melanoma cells, in part by altering the amount and nature of the GAG molecules produced.Glycosaminoglycans (GAGs) are long-chain polyanionic carbohydrates that include hyaluronic acid, chondroitin 4-and 6-sulfates, keratan sulfate, dermatan sulfate, heparin, and heparan sulfate (1). These macromolecules are found in most mammalian cells (2) and are believed to play an important role in migration (3-6), maintenance of morphogenetic structure (7), and cellular differentiation (8, 9). Despite intensive study, little is known about the factors that regulate the biosynthesis of these polysaccharides. However, previous studies have suggested that the extracellular environment can influence the rate of synthesis and secretion of these macromolecules (10, 11).Collagen is a major constituent of the extracellular matrix. Type I collagen is found predominently in bone, dermis, and tendon, whereas type IV collagen is found in basement membranes (12). Various types of collagen bind to GAGs both in their native proteoglycan state and in the form of polysaccharide chains (13)(14)(15)(16)(17)(18). For this reason, the coordinated biosynthesis of collagen and proteoglycan has been investigated (19-21); both of these kinds of macromolecules appear to be produced in the same cytoplasmic region (22) A collagen-mediated reduction in the rate of cell-associated GAG degradation rather than an actual change in the rate of synthesis of these macromolecules has been postulated (26, 27). However, mammary epithelial cells that have spontaneously transformed to a malignant phenotype are less effective than normal cells in decreasing proteoglycan degradation in response to collagen (29). Other malignant lines have been reported to show alterations in both the synthesis of cell surface proteins and their interaction with matrix proteins (30, 31). Because malignant melanoma cells are exposed to type I collagen during local invasion into the dermis and are in contact with type IV collagen during metastatic sprea...

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Section: Resultsmentioning

confidence: 99%

Influence of collagen substrata on glycosaminoglycan production by B16 melanoma cells.

Luikart

Maniglia

Sartorelli

1983

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

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“…Since monoclonal antibodies B3 and 33 clearly recognize different antigens (see also immunocytochemical observations), it is most likely that a small proportion of each antigen exists as a complex in conditioned medium and can thus be precipitated by either antibody. This is not entirely unexpected since antigen-33 appears to be a heparan sulfate proteoglycan (see below) and fibronectin is known to bind both heparin and heparan sulfate (29,38,49). (Antibody-33 does not recognize SDS-denatured antigen and could not be used in Western blots.)…”

Section: Antigen-31--lamininmentioning

confidence: 99%

Extracellular matrix organization in developing muscle: correlation with acetylcholine receptor aggregates.

Bayne¹,

Anderson²,

Fambrough³

1984

The Journal of Cell Biology

177

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Monoclonal antibodies recognizing laminin, heparan sulfate proteoglycan, fibronectin, and two apparently novel connective tissue components have been used to examine the organization of extracellular matrix of skeletal muscle in vivo and in vitro. Four of the five monoclonal antibodies are described for the first time here.Immunocytochemical experiments with frozen-sectioned muscle demonstrated that both the heparan sulfate proteoglycan and laminin exhibited staining patterns identical to that expected for components of the basal lamina. In contrast, the remaining matrix constituents were detected in all regions of muscle connective tissue: the endomysium, perimysium, and epimysium.Embryonic muscle cells developing in culture elaborated an extracellular matrix, each antigen exhibiting a unique distribution. Of particular interest was the organization of extracellular matrix on myotubes: the build-up of matrix components was most apparent in plaques overlying clusters of an integral membrane protein, the acetylcholine receptor (AChR). The heparan sulfate proteoglycan was concentrated at virtually all AChR clusters and showed a remarkable level of congruence with receptor organization; laminin was detected at 70-95% of AChR clusters but often was not completely co-distributed with AChR within the cluster; fibronectin and the two other extracellular matrix antigens occurred at ~20, 8, and 2% of the AChR clusters, respectively, and showed little or no congruence with AChR. From observations on the distribution of extracellular matrix components in tissue cultured fibroblasts and myogenic cells, several ideas about the organization of extracellular matrix are suggested. (a) Congruence between AChR clusters and heparan sulfate proteoglycan suggests the existence of some linkage between the two molecules, possibly important for regulation of AChR distribution within the muscle membrane. (b) The qualitatively different patterns of extracellular matrix organization over myotubes and fibroblasts suggest that each of these cell types uses somewhat different means to regulate the assembly of extracellular matrix components within its domain. (c) The limited co-distribution of different components within the extracellular matrix in vitro and the selective immune precipitation of each antigen from conditioned medium suggest that each extracellular matrix component is secreted in a form that is not complexed with other matrix constituents.The surface of adult vertebrate skeletal muscle fibers is regionally specialized, both at tendon-attachment sites and at the neuromuscular junction. While the mechanisms that control the formation and maintenance of such specialized regions are still not clearly understood, attention has been focused recently on the possible influence of extracellular matrix on muscle surface organization. The extracellular matrix surrounding each muscle fiber can be divided into two obvious morphological layers (see reference 40). Most proximal to the sarcolemma is a felt-like band, -10-15-nm thick,

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“…The supematant was dialyzed extensively against PBS and passed through a heparin-Sepharose affinity column which was washed sequentially with PBS. 0.5 M NaC1 (in 50 mM Tris, pH 7.4), which elutes quantitatively any contaminating FN (24), l M NaCI (in 50 mM Tris), and finally 2 M NaC1 (in 50 mM Tris) to elute the bound PF4. The identity and purity of the recovered protein was verified by amino acid analysis (12) on a Durrum 500 amino acid analyzer and by polyacrylamide gel electrophoresis.…”

Section: Methodsmentioning

confidence: 99%

“…Fibronectin consists of dimeric subunits (54) and has the potential to interact with several cell surfaceassociated macromolecules including collagen (l 3), certain glycosaminoglycans (24,56), and possibly gangliosides (21), and another unidentified cell surface receptor (37,43,45,56). Accumulating evidence indicates that cell surface glycosaminoglycans and proteoglycans mediate fibroblast attachment and possibly more complex adhesive responses on fibronectincontaining extracellular matrices (10,41,47).…”

mentioning

confidence: 99%

Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin.

Laterra¹,

Silbert²,

Culp³

1983

The Journal of Cell Biology

Self Cite

268

143

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Proteins with affinities for specific glycosaminoglycans (GAG's) were used as probes for testing the potential of cell surface GAG's to mediate cell adhesive responses to extracellular matrices (ECM). Plasma fibronectin (FN) and proteins that bind hyaluronate (cartilage proteoglycan core and link proteins) or heparan sulfate (platelet factor 4 [PF4]) were adsorbed to inert substrata to evaluate attachment and spreading of several 3T3 cell lines. Cells failed to attach to hyaluronate-binding substrata. The rates of attachment on PF4 were identical to those on FN; however, PF4 stimulated formation of broad convex lamellae but not tapered cell processes fibers during the spreading response. PF4-mediated responses were blocked by treating the PF4-adsorbed substratum with heparin (but not chondroitin sulfate), or alternatively the cells with Flavobacter heparinum heparinase (but not chondroitinase ABC). Heparinase treatment did not inhibit cell attachment to FN but did inhibit spreading. Cells spread on PF4 or FN contained similar Ca2+-independent cell-substratum adhesions, as revealed by EGTA-mediated retraction of their substratum-bound processes. Microtubular networks reorganized in cells on PF4 but failed to extend into the broadly spread lamellae, where fine microfilament bundles had developed. Stress fibers, common on FN, failed to develop on PF4. These experiments indicate that (a) heparan sulfate proteoglycans are critical mediators of cell adhesion and heparan sulfate-dependent adhesion via PF4 is comparable in some, but not all, ways to FN-mediated adhesion, (b) the uncharacterized and heparan sulfate-independent "cell surface" receptor for FN permits some but not all aspects of adhesion, and (c) physiologically compatible and complete adhesion of fibroblasts requires binding of extracellular matrix FN to both the unidentified "cell surface" receptor and heparan sulfate proteoglycans.Fibronectin is an extracellular matrix-associated glycoprotein found in vivo that mediates fibroblast attachment, spreading, motility, and longterm survival in vitro when adsorbed to tissue culture substrata. Fibronectin consists of dimeric subunits (54) and has the potential to interact with several cell surfaceassociated macromolecules including collagen (l 3), certain glycosaminoglycans (24, 56), and possibly gangliosides (21), and another unidentified cell surface receptor (37,43,45,56). Accumulating evidence indicates that cell surface glycosaminoglycans and proteoglycans mediate fibroblast attachment and possibly more complex adhesive responses on fibronectincontaining extracellular matrices (10, 41,47). Hyaluronic acid and heparan sulfate are two particular classes of cell surface glycosaminoglycan that have received considerable attention as potential determinants of the state of cellular adhesiveness and motility in numerous experimental systems (2, I l, 25, 34, 44). It has yet to be determined, however, if direct interactions between cell surface glycosaminoglycans and the extracellular matrix determine prop...

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Glycosaminoglycans that bind cold-insoluble globulin in cell-substratum adhesion sites of murine fibroblasts.

Cited by 102 publications

References 26 publications

Influence of collagen substrata on glycosaminoglycan production by B16 melanoma cells.

Influence of collagen substrata on glycosaminoglycan production by B16 melanoma cells.

Extracellular matrix organization in developing muscle: correlation with acetylcholine receptor aggregates.

Cell surface heparan sulfate mediates some adhesive responses to glycosaminoglycan-binding matrices, including fibronectin.

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