Basement membrane glycoprotein laminin binds to heparin

Sakashita, Shigeo; Engvall, Eva; Ruoslahti, Erkki

doi:10.1016/0014-5793(80)80654-5

Cited by 277 publications

(114 citation statements)

References 16 publications

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“…Because fibronectin codistributes with laminin, collagen, and heparan sulfate proteoglycan when extracellular matrix is examined by immunofluorescence microscopy (31)(32)(33)(34), thrombospondin may also codistribute with these proteins. Thrombospondin, laminin, and fibronectin all interact with heparin (9,28,35). In addition, the binding of fibronectin to collagen is enhanced by glycosaminoglycans, and the complexes formed by these interactions are insoluble (36)(37)(38).…”

Section: Discussionmentioning

confidence: 99%

Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix.

Jaffe¹,

Ruggiero²,

Leung³

et al. 1983

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

254

125

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Thrombospondin, a major glycoprotein released from a granules of thrombin-stimulated platelets, is a disulfidebonded trimer of 160-kilodalton subunits. Cultured human foreskin and fetal lung fibroblasts secreted thrombospondin (determined by enzyme-linked immunosorbent assay) into the culture medium in a time-dependent manner (15.7 and 5.8 Thrombospondin is a major platelet a granule glycoprotein that is secreted and then partially bound to platelet membranes when human platelets aggregate in response to thrombin (1)(2)(3)(4)(5)(6)(7)(8). Thrombospondin is a 450-kilodalton (kDal) protein and is composed of three large disulfide-linked subunits (9, 10). During platelet aggregation, thrombin-stimulated platelets develop a membrane-bound lectin-like activity (11-13), which originates from a granules and appears to play a role in mediating platelet aggregation by binding to a specific receptor on other platelets (14,15 3.7 X 10'°Bq). The radioactive postculture medium was removed, centrifuged at 8,000 x g for 2 min to remove cells and debris, and frozen at -35°C. The radioactive cell layers were washed twice, removed with a rubber policeman, pelleted by centrifugation, dissolved by boiling for 5 min in 2% NaDodSO4 containing protease inhibitors as described (20), and frozen until analyzed.In experiments in which the accumulation of thrombospondin antigen was measured by enzyme-linked immunosorbent assay (ELISA), fibroblasts were cultured in minimal essential medium containing 20% rabbit serum in 2-cm2 wells of multiwell plates. When the cells were confluent, the cells were washed, and the medium was replaced with 1 ml of fresh minimal essential medium containing 20% rabbit serum. At various times after the medium change, the postculture medium was removed, centrifuged at 8,000 X g for 2 min, and frozen until assayed. 998The publication costs ofthis article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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Section: Discussionmentioning

confidence: 99%

Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix.

Jaffe¹,

Ruggiero²,

Leung³

et al. 1983

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

254

125

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“…Proteoglycans are known to bind to several components of the extracellular matrix, including fibronectin (50, 53,67), laminin (52,65), and collagen (32,53,54), and also other proteoglycans (15). It has also been shown that a membrane-intercalated heparan sulfate proteoglycan isolated from mammary epithelial cells may interact with polymerized actin (47).…”

mentioning

confidence: 99%

The melanoma proteoglycan: restricted expression on microspikes, a specific microdomain of the cell surface.

Garrigues

Lark

Lara

et al. 1986

The Journal of Cell Biology

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Abstract.A cell surface chondroitin sulfate proteoglycan associated with human melanomas and defined by mAb's F24.47 and 48.7 has been characterized biochemically and localized by indirect immunogold electron microscopy. These antibodies recognize distinct epitopes on the intact proteoglycan. In addition, mAb 48.7 also recognizes an epitope on a 250,000-D glycoprotein and is therefore similar to antibody 9.2.27 (described by Bumol, T. E, and R. A. Reisfeld, 1982, Proc. Natl. Acad. Sci. USA., 79:1245-1249. Furthermore, it was shown that the glycosaminoglycan chains released by alkaline borohydride treatment of the proteoglycan recognized by mAb 48.7 had a size of '~60,000 D. Since the intact proteoglycan was estimated to be 420,000 D, there are probably three chondroitin sulfate chains attached to the 250,000-D core glycoprotein. Furthermore, an oligosaccharide fraction containing 42 % of the 3H activity (glucosamine as precursor) was isolated.Immunolocalization studies using whole-mount electron microscopy revealed that the chondroitin sulfate proteoglycan was present almost exclusively on microspikes, a microdomain of the melanoma cell surface. These processes were present as 1-2-~tm structures on the upper cell surface and as longer (up to 20 ~tm) structures at the cell periphery. Peripheral microspikes were involved in the initial interactions between adjacent cells and formed complex footpads that made contact with the substratum. Immunogold-labeled cells were also thin sectioned and the specific localization of the chondroitin sulfate proteoglycan antigen was quantitated. The data confirmed the results of wholemount microscopy and demonstrated a statistically significant association of the antigen with the microspike processes as compared with other areas of the cell surface. By using two different mAb's (48.7 and F24.47) that recognize epitopes on either the core glycoprotein or the intact proteoglycan, respectively, we have demonstrated that both molecules have the same restricted distribution at the cell surface. The specific localization of the antigen to microspikes at the cell surface suggests it may play a role in cell-cell contact and cell-substratum adhesion, which could be important in the metastatic process.

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“…Dr . Chung provided both whole rabbit antiserum and affinitypurified antibody to a protein, GP2, isolated from extracellular matrix secreted by the mouse embryonal carcinoma-derived endodermal cell line M 1536-B3 (9) ; GP-2 has been shown to be a subunit of laminin (34,46). All three preparations were studied by immunotluorescence (Fig.…”

mentioning

confidence: 99%

Laminin, fibronectin, and collagen in synaptic and extrasynaptic portions of muscle fiber basement membrane.

Sanes¹

1982

The Journal of Cell Biology

254

124

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Light and electron microscope immunohistochemical methods were used to study the distribution of several proteins in rat skeletal muscle . The aims were to identify components of muscle fiber basement membrane and to compare the small fraction (0 .1%) of the basement membrane that extends through the synaptic cleft at the neuromuscular junction with the remaining, extrasynaptic portion . Synaptic basement membrane is functionally specialized and plays important roles in neuromuscular function and regeneration . Laminin, fibronectin, Collagen IV, Collagen V, and a collagenous protein (high-salt-soluble protein [HSP]) are all present in muscle fiber basement membrane . Laminin and Collagen IV are concentrated in basal lamina (the feltlike, inner layer of the basement membrane) and are shared by synaptic and extrasynaptic regions . Fibronectin, also present synaptically and extrasynaptically, is present in basal lamina and in the overlying reticular lamina . Collagen V and HSP are present throughout extrasynaptic basement membrane but are absent from synaptic sites; HSP is concentrated in the reticular lamina and on the outer surface of the basal lamina . These results, together with experiments reported previously (Sanes and Hall, 1979, /. Cell Biol. 83 :357-370), provide examples of three classes of components in muscle fiber basement membranesynaptic, extrasynaptic, and shared .Each muscle fiber in vertebrate skeletal muscles is ensheathed by a basement membrane . A small fraction of the basement membrane, -0.1% of the total, occupies the synaptic cleft between nerve and muscle at the neuromuscular junction. Recent experiments have demonstrated that this synaptic portion of the basement membrane is specialized and plays important roles in neuromuscular function and regeneration (1,2,5,19,28,36) . These results motivate molecular analysis of muscle-fiber basement membrane and comparison of its synaptic and extrasynaptic portions. In a previous study, we used immunohistochemical methods to show that several antigens are present in synaptic but not extrasynaptic basement membrane (35). Here, to extend this analysis to extrasynaptic areas, we asked two questions : (a) Are any proteins that have been purified and characterized from other sources present in rat muscle fiber basement membrane? And (b) are proteins which are present in extrasynaptic basement membrane also present at the synapse or are there antigens specific to extrasynaptic regions? We found that three proteins recently shown to be components of basement membranes in several tissues-laminin (9,13,25,32,37,46), collagen IV (4,33,38,45,51), and fibronectin (10,26,29,42,50)-are present in synaptic as well as extrasynaptic muscle fiber basement membrane . Two other proteins-collagen V (4,6,11,15,16,31,33) and a collagenous protein (HSP) isolated from skeletal muscle connective tissueare present in extrasynaptic basement membrane but are excluded from synaptic sites . MATERIALS AND METHODS AntibodiesLAMININ : Antibodies to laminin were provided by ...

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Basement membrane glycoprotein laminin binds to heparin

Cited by 277 publications

References 16 publications

Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix.

Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix.

The melanoma proteoglycan: restricted expression on microspikes, a specific microdomain of the cell surface.

Laminin, fibronectin, and collagen in synaptic and extrasynaptic portions of muscle fiber basement membrane.

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