Isolation and characterization of a high molecular weight type IV collagenase isolated from human carcinoma tissue.
A proform of high molecular weight type IV collagenase was isolated and purified 1230-fold from human metastatic carcinoma tissue. Like matrix metalloproteinases (MMPs), the enzyme was activated by trypsin and degraded type IV collagen and gelatin at a neutral pH, the activity was inhibited by EDTA and o-phenanthroline. However, the molecular weight was much higher than MMPs which degraded type IV collagen, gelatinase A (MMP-2; 72 kDa gelatinase/type IV collagenase) (EC 3.4.24.24), gelatinase B (MMP-9; 92 kDa gelatinase/type IV collagenase) (EC 3.4.24.35), stromelysin-1 (MMP-3; 57 kDa) (EC 3.4.24.17) and stromelysin-2 (MMP-10; 57 kDa) (EC 3.4.24.22). The other significant difference from MMPs was that the enzyme was not activated by 4-aminophenylmercuric acetate nor inhibited by TIMP. Taking together these results, this high molecular weight type IV collagenase might be a newly found enzyme different from MMPs or might have the same configuration as MMPs already reported.
Tsuda
T T
TT
Molecular Life Science, School of Medicine, Tokai University, Kanagawa, Japan.
Kodama
A
A
Yamamura
M
M
Matsuzaki
S
S
Tsuda
M
M
D003094:Collagen / Q000378:metabolism; D017364:Collagenases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases; D006023:Glycoproteins / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008113:Liver Neoplasms / Q000201:enzymology* / Q000556:secondary; D020780:Matrix Metalloproteinase 9; D008666:Metalloendopeptidases / Q000737:chemistry / Q000378:metabolism; D008970:Molecular Weight; D010434:Pepsin A / Q000302:isolation & purification; D010618:Phenanthrolines / Q000494:pharmacology; D013379:Substrate Specificity; D019714:Tissue Inhibitor of Metalloproteinases; D014357:Trypsin / Q000494:pharmacology
D006023:Glycoproteins; D010618:Phenanthrolines; D019714:Tissue Inhibitor of Metalloproteinases; D005780:Gelatin; D003094:Collagen; D004492:Edetic Acid; D014357:Trypsin; D010434:Pepsin A; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; D020780:Matrix Metalloproteinase 9; C025205:1,10-phenanthroline