Characterisation of chondroitin/dermatan sulphate proteoglycans synthesised by rat mammary myoepithelial and fibroblastic cell lines.
Chondroitin sulphate proteoglycans were isolated from the culture medium of rat mammary gland fibroblast (Rama 27) and myoepithelial (Rama 401) cell lines which had been labelled with [35S]sulphate. Chromatography on Sepharose CL-4B indicated that the Rama 401 proteoglycan was larger than the Rama 27 proteoglycan (Kav values 0.47 and 0.56, respectively). Treatment of the proteoglycans with alkaline NaBH4 yielded chondroitin sulphate chains with average M(r) values of 37,000 (Rama 401) and 21,000 (Rama 27). Structural analysis of the glycosaminoglycan chains indicated that both were co-polymers of chondroitin and dermatan sulphate although there were differences in the amounts and distribution of the disaccharide repeating units. The M(r) values of the core proteins, determined by immunoblotting, were about 43,000 and 46,000 (Rama 27) and 44,500 (Rama 401). Using an antibody to chondroitin sulphate proteoglycan in immunofluorescence experiments, the proteoglycan was demonstrated on the surface of both cell lines. Rama 27 cells additionally possessed an extensive fibrous extracellular matrix which also stained with the antibody. Staining of sections of lactating mammary gland suggested that the proteoglycan was present in the basement membrane as well as the stromal connective tissue. The presence of chondroitin sulphate proteoglycan in the basement membrane was confirmed by ultrastructural immunolocalisation.
Warburton
M J
MJ
Department of Histopathology, St George's Hospital Medical School, London, UK.
Kimbell
R
R
Ferns
S A
SA
Hayman
A R
AR
Perusinghe
N
N
Monaghan
P
P
D000818:Animals; D001704:Biopolymers; D002240:Carbohydrate Sequence; D002460:Cell Line; D002809:Chondroitin Sulfates / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D003871:Dermatan Sulfate / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D004848:Epithelium / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011509:Proteoglycans / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification*; D051381:Rats; D013462:Sulfur Radioisotopes
D001704:Biopolymers; D011509:Proteoglycans; D013462:Sulfur Radioisotopes; C069524:dermatan sulfate chondroitin sulfate; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates