Identification and characterization of a novel matrix-degrading protease from hormone-dependent human breast cancer cells.
A novel matrix-degrading enzyme was identified from human breast cancer cells. This enzyme appears as major gelatinase in hormone-dependent breast cancer cell lines and has as an apparent molecular mass of 80 kDa on gelatin zymography. The 80-kDa enzyme has a unique metal ion specificity. In addition to calcium ions, the gelatinolytic activity can be supported by manganese and/or magnesium. Unlike 92- and 72-kDa gelatinases and other known members of the metalloproteinase family, the 80-kDa protease is not activated by p-aminophenylmercuric acetate and its gelatinolytic activity is not inhibited by tissue inhibitor of metalloproteinase 2. It is active over the pH range 7.5-9.5 with an optimum at pH 8.5. The enzyme degrades gelatin and type IV collagen. The proteolytic activity of the enzyme is inhibited by EDTA and leupeptin. These unique features clearly distinguish the 80-kDa protease from the known 92-and 72-kDa gelatinases. The expression of 80-kDa enzyme can be detected in hormone-dependent human breast cancer cell lines in vitro and in tumors grown from these cells in athymic nude mice.
Vincent T. Lombardi Cancer Research Center, Georgetown University Medical Center, Washington, D.C. 20007.
Y E
YE
Shi
J
J
Torri
L
L
Yieh
A
A
Wellstein
M E
ME
Lippman
R B
RB
Dickson
D000595:Amino Acid Sequence; D001943:Breast Neoplasms / Q000201:enzymology*; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D005260:Female; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000494:pharmacology; D009376:Neoplasms, Hormone-Dependent / Q000201:enzymology*; D010434:Pepsin A / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives / Q000494:pharmacology; D013379:Substrate Specificity; D019716:Tissue Inhibitor of Metalloproteinase-2; D014407:Tumor Cells, Cultured
D009363:Neoplasm Proteins; D019716:Tissue Inhibitor of Metalloproteinase-2; C010024:4-aminophenylmercuriacetate; D005780:Gelatin; D004492:Edetic Acid; D010450:Endopeptidases; D010434:Pepsin A; D018093:Gelatinases; D010662:Phenylmercuric Acetate