Mitogenic activity of procathepsin D purified from conditioned medium of breast-cancer cells by affinity chromatography on pepstatinyl agarose.
A simple procedure for the affinity purification of procathepsin D from tissue culture medium conditioned by breast-cancer cells is described. This procedure yielded 2 micrograms of procathepsin D/100 ml medium. The procathepsin D was approximately 95% pure as judged by silver staining of polyacrylamide gels, the major contaminant being mature cathepsin D. The ability of procathepsin D to stimulate the proliferation of oestrogen-responsive MCF-7 breast cancer cells was determined. The purified procathepsin D had no mitogenic effect alone or in combination with oestradiol or other growth factors. These data suggest that procathepsin D does not act as an oestrogen-regulated autocrine growth factor for malignant breast epithelial calls.
Stewart
A J
AJ
Department of Pathology, Royal Victoria Infirmary, Newcastle upon Tyne, UK.
Piggott
N H
NH
May
F E
FE
Westley
B R
BR
D001943:Breast Neoplasms / Q000737:chemistry / Q000473:pathology*; D002402:Cathepsin D / Q000302:isolation & purification / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002846:Chromatography, Affinity; D004792:Enzyme Precursors / Q000302:isolation & purification / Q000494:pharmacology*; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008934:Mitogens; D010436:Pepstatins; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; D008934:Mitogens; D010436:Pepstatins; C014195:Streptomyces pepsin inhibitor; D004958:Estradiol; C058345:procathepsin D; D002402:Cathepsin D; C031375:pepstatin