Gelatin-degrading type IV collagenase isolated from human small cell lung cancer.
The goal of this study has been to identify and characterize metalloproteinases from a highly metastatic human small cell lung cancer cell line. The cytosol isolated from NCI-H82 lung cancer cells propagated as solid tumors in nude mice contained a gelatinolytic enzyme that was subjected to ammonium sulfate precipitation, zinc chelate Sepharose column chromatography, anion exchange chromatography and gel permeation chromatography. This purification scheme resulted in a 280-fold enrichment of an active gelatin and type IV collagen-degrading enzyme. On gelatin zymography two bands of gelatinolytic activity were detected, corresponding to Mr of 75,000 and 63,000. Gelatinolytic activity was inhibited by metal chelators, tetracyclines, and serum. On immunoblotting using an affinity-purified polyclonal rabbit antibody to a peptide region of type IV collagenase, the tumor enzyme was identified as type IV collagenase. A second tumor metalloproteinase of Mr = 29,000, which degraded proteoglycan substrates, was also isolated.
Zucker
S
S
Department of Research, Veterans Administration Medical Center, Northport, N.Y.
Wieman
J
J
Lysik
R M
RM
Imhof
B
B
Nagase
H
H
Ramamurthy
N
N
Liotta
L A
LA
Golub
L M
LM
D000818:Animals; D018288:Carcinoma, Small Cell / Q000201:enzymology*; D003094:Collagen / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D008175:Lung Neoplasms / Q000201:enzymology*; D051379:Mice; D008819:Mice, Nude; D008970:Molecular Weight; D009368:Neoplasm Transplantation; D010434:Pepsin A / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D005780:Gelatin; D003094:Collagen; D010434:Pepsin A; D018093:Gelatinases