Colocalization of a large heterodimeric proteoglycan with basement membrane proteins in cultured cells.
A novel large heterodimeric dermatan sulfate proteoglycan with core proteins of 460 and 300 kDa, respectively, had been described as a secretory product of human fetal skin fibroblasts (Breuer et al., J. Biol. Chem. 266, 13224-13232 (1991)). Pulse-chase experiments showed a preferential association of the proteoglycan with the cell membrane. Immunogold labeling indicated its localization in fibrils on the cell surface as well as in fibrillar extensions from the cell body. Immunofluorescence studies yielded a fibrillar and punctate staining pattern which was also seen in cultured human and porcine endothelial cells. Dot-like structures were observed in transformed human keratinocytes. Various immunocytochemical double-labeling experiments indicated a remarkable colocalization of the proteoglycan with fibronectin, laminin, perlecan, and type IV collagen whereas only occasionally a colocalization with chondroitin-6-sulfate was found. No evidence for an enrichment of the proteoglycan in vinculin-containing structures was obtained. These results suggest that the proteoglycan is a widely distributed macromolecule which can associate with basement membrane components. Preliminary findings in rat cornea supported this conclusion.
Faber
V
V
Institute of Arteriosclerosis Research, University of Münster, Germany.
Quentin-Hoffmann
E
E
Breuer
B
B
Schittny
J
J
Völker
W
W
Kresse
H
H
D001485:Basement Membrane / Q000737:chemistry*; D002478:Cells, Cultured / Q000737:chemistry; D003094:Collagen / Q000737:chemistry; D003315:Cornea / Q000737:chemistry*; D004730:Endothelium, Vascular / Q000737:chemistry; D005347:Fibroblasts / Q000737:chemistry*; D005353:Fibronectins / Q000737:chemistry; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry; D007797:Laminin / Q000737:chemistry; D011509:Proteoglycans / Q000032:analysis* / Q000737:chemistry
D003094:Collagen; D005353:Fibronectins; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D007797:Laminin; D011509:Proteoglycans; C071927:perlecan