Isolation and characterization of basement membrane and cell proteoheparan sulphates from HR9 cells

Abstract

The mouse teratocarcinoma cell line HR9 was investigated for proteoheparan sulphate production. Four species of proteoheparan sulphate molecules were isolated and purified to homogeneity. The proteoheparan sulphate isolated from the tissue-culture medium contains four heparan sulphate side-chains of 25 kDa each, and its core protein has an approximate molecular mass of 50 kDa. The proteoheparan sulphates associated with the cells were separated into three individual species: cell proteoheparan sulphate I exhibits structural characteristics which are very similar to the proteoheparan sulphate isolated from the tissue culture medium; cell proteoheparan sulphates II and III contain one heparan sulphate chain of 25 kDa and 20 kDa, and core proteins of approximately 30 kDa and 25 kDa respectively. Antisera, raised against the medium form, react specifically with basement membranes in various tissues by immunofluorescence. This staining pattern was compared to the pattern observed with an antiserum which we have obtained to a proteoheparan sulphate species isolated from the plasma membrane of bovine aortic endothelial cells. The structural and immunological data suggest that basement membrane and plasma membrane proteoheparan sulphates are different biosynthetic products and are not directly related to each other.