Collagen-binding proteins of rat mammary tumor epithelial cells: a biochemical and immunological study.
Collagen-binding proteins were studied in mammary epithelial cells of 7,12-dimethylbenz[a]anthracene-induced rat mammary tumors. These proteins can be solubilized from cell membranes with 0.1% Triton. Using affinity chromatography on type I collagen-Sepharose and polyacrylamide slab gel electrophoresis, three major proteins of 34,000, 36,000, and 38,000 Da were found. Similar proteins were also present in several other cell types, including both epithelial and mesenchymal cells. Pulse-chase experiments did not indicate a precursor-product relationship of these proteins. Tryptic/chymotryptic peptide maps, however, revealed that the 36,000- and 38,000-Da proteins are very similar but are quite different from the 34,000-Da molecular form. The distribution and function of these proteins were then analyzed by using polyclonal antibodies directed against the entire set of major proteins. In immunofluorescence studies we observed a dense, punctate distribution of fluorescence on the cell surface of isolated and unfixed epithelial organoids and a bright pericellular staining in cultures after fixation. Treatment with the antiserum did not affect attachment and spreading of cuboidal mammary cells to plastic or to a collagen substratum. However, when the antiserum was added to the medium of growing cuboidal cells, it caused the formation of duct-like structures. These studies indicate that collagen-binding proteins may play a role in mammary gland morphology.
Wirl
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Institut für Molekularbiologie der Osterreichischen Akademie der Wissenschaften, Salzburg, Austria.
Pfäffle
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D000818:Animals; D002448:Cell Adhesion; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism*; D046508:Culture Techniques; D004848:Epithelium / Q000378:metabolism / Q000473:pathology; D005455:Fluorescent Antibody Technique; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism* / Q000473:pathology; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D010449:Peptide Mapping; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D038981:Receptors, Collagen
D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D003094:Collagen