Heparan sulfate proteoglycan of cultured cells: Demonstration of a lipid- and a matrix-associated form

Norling, Börje; Glimelius, Bengt; Wasteson, Åke

doi:10.1016/0006-291x(81)90259-x

Cited by 89 publications

(32 citation statements)

References 19 publications

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“…A class of cell surface heparan sulfate has been shown to exist covalently linked to protein in a proteoglycan form (22,36). Certain heparan sulfate proteoglycans insert into lipid bilayers via a hydrophobic core protein and may exist as integral membrane components on the surface of the cell (20,30,35). These plasma membrane forms of heparan proteoglycan should be so tightly bound to the cell surface that they would mediate at least simple cytoplasmic attachment responses to heparan sulfate-binding matrices such as that provided by PF4 or FN.…”

Section: Discussionmentioning

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

267

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...

show abstract

Section: Discussionmentioning

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

267

143

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“…Proteoheparan sulphates from hepatocytes (Kjellen et al, 1981), glial cells and fibroblasts (Norling et al, 1981) as well as epithelial cells (Rapraeger & Bernfield, 1983) have been found to be membrane-associated. The proteoglycan studied in the present work is presumably also a membraneassociated form.…”

Section: Discussionmentioning

confidence: 99%

The core protein of fibroblast proteoheparan sulphate consists of disulphide-bonded subunits

Cöster¹,

Malström²,

Carlstedt³

et al. 1983

Biochemical Journal

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Fibroblast proteoheparan sulphate has a disulphide-bonded subunit structure. The core protein appears to consist of two polypeptides each of Mr 80 000-100 000. As shown elsewhere [Carlstedt, Cöster, Malmström & Fransson (1983) J. Biol. Chem. in the press], both polypeptide molecules carry four to six heparan sulphate side chains (approx. Mr 20 000) and an unknown number of oligosaccharide units, giving the whole macromolecule an Mr in the range 300 000-400 000.

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“…(Branford White and Hardy, 1977, Kanwar, et al 1981, Kraemer, 1971, Mutoh, et al, 1978, Norling, et al, 1981and Oldberg, et al, 1979. The location of heparin or heparan sulfate proteoglycans in the thyroid tissue remains to be studied.…”

Section: Discussionmentioning

confidence: 99%

Presence of heparan sulfate proteoglycan in thyroid tissue.

Shishiba

Yanagishita

1983

Endocrinol Japon

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Glycosaminoglycan (GAG) was extracted from the porcine thyroid gland with a buffer comprising 5.3 M guanidine-HCl and proteolytic enzyme inhibitors and was fractionated by subsequent isodensity CsCl centrifugation.60% of uronic acid positive materials was accumulated in the bottom one-fourth fraction with high buoyant density. More than 90% of this uronic acid positive material in the thyroid tissue was heparin or heparan sulfate (sensitive to nitrous acid treatment) and the rest was chondroitin sulfate or dermatan sulfate (sensitive to chondroitinase ABC treatment). When the accumulated high buoyant density GAG was analyzed on a Sepharose CL-6-B column, approximately 14% of the heparin sulfate were in the macromolecular portion as a form of proteoglycan because it was destroyed by the papain digestion or alkaline borohydride treatment which extensively digests protein or releases GAG from protein by the elimination reaction, respectively.This study demonstrates the existence of heparan sulfate proteoglycan in thyroid tissue for the first time.

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Heparan sulfate proteoglycan of cultured cells: Demonstration of a lipid- and a matrix-associated form

Cited by 89 publications

References 19 publications

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

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

The core protein of fibroblast proteoheparan sulphate consists of disulphide-bonded subunits

Presence of heparan sulfate proteoglycan in thyroid tissue.

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