Human breast milk contains procathepsin D--detection by specific antibodies.
The presence of the zymogen of cathepsin D in human milk was detected using antibodies specific for the proenzyme and by the proteolytic activity at low pH. The antibodies were raised against a synthetic propeptide of human cathepsin D and were tested using immunoprecipitations and western blots of samples from different breast cancer cell lines as well as cytosol fractions of human breast cancer tissues. In all experiments these antibodies recognized specifically procathepsin D. Procathepsin D from human milk was partially activated at low pH. The activity was monitored using hemoglobin 14C proteolytic assay, and it was abolished by pepstatin A--a specific inhibitor of aspartic proteinases. Western blots did not reveal presence of cathepsin B or cathepsin H. These data indicate specific secretion of cathepsin D in human breast milk.
Vĕtvicka
V
V
Department of Microbiology and Immunology, University of Louisville, KY 40292.
Vágner
J
J
Baudys
M
M
Tang
J
J
Foundling
S I
SI
Fusek
M
M
D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000201:enzymology; D002402:Cathepsin D / Q000032:analysis* / Q000276:immunology; D003470:Culture Media; D004792:Enzyme Precursors / Q000032:analysis* / Q000276:immunology; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008895:Milk, Human / Q000201:enzymology*; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured
D000906:Antibodies; D003470:Culture Media; D004792:Enzyme Precursors; C058345:procathepsin D; D002402:Cathepsin D