Structure of the pericellular matrix: association of heparan and chondroitin sulfates with fibronectin-procollagen fibers.
Immunofluorescent staining of a pericellular matrix produced by cultured human embryonic skin fibroblasts showed a codistribution among fibronectin, heparan sulfate proteoglycans and part of the chondroitin sulfate in a fibrillar network. Isolated matrix in an "intact" form could be scraped off the dish after detergent solubilization of the cells. On centrifugation in cesium chloride density gradients, most sulfated glycosaminoglycans and matrix proteins remained associated and were recovered at a density of 1.34 g/cm3 (greater or equal to 2 M CsCl). However, when 4 M guanidine hydrochloride was included in the gradient medium, the components dissociated, suggesting that the sulfated glycosaminoglycans are bound to matrix proteins by strong noncovalent linkages. Interactions between sulfated glycosaminoglycans produced by the fibroblasts and fibronectin could also be demonstrated by affinity chromatography on immobilized plasma fibronectin and by immunoprecipitation of fibronectin in conditioned culture medium, which resulted in a coprecipitation of the sulfated glycosaminoglycans. In these two systems, the fibronectin glycosaminoglycan bonds were broken at 0.2 M salt and were apparently weaker than the bonds responsible for the structural integrity of the matrix. These findings implicate heparan and chondroitin sulfate proteoglycans as integral compounds of the pericellular matrix fibers and suggest that the association of the proteoglycans with the fibronectin-procollagen matrix is stabilized by multiple molecular interactions.
D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D002807:Chondroitin / Q000031:analogs & derivatives*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D002846:Chromatography, Affinity; D003470:Culture Media / Q000032:analysis; D005347:Fibroblasts / Q000648:ultrastructure; D005353:Fibronectins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D012867:Skin / Q000196:embryology; D012965:Sodium Chloride / Q000494:pharmacology
D002807:Chondroitin; D002809:Chondroitin Sulfates; D003470:Culture Media; D005353:Fibronectins; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D012965:Sodium Chloride