Isolation and characterization of a high molecular weight type IV collagenase isolated from human carcinoma tissue

• Published in: FEBS letters 1993-03, Vol.319 (1), p.35-39
• Main Authors: Tsuda, Tomi T., Kodama, Akira, Yamamura, Masaichi, Matsuzaki, Shohei, Tsuda, Michio
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; APMA, 4-aminophenylmercuric acetate; Biological and medical sciences; Collagen - metabolism; Collagenases - chemistry; Collagenases - isolation & purification; Collagenases - metabolism; DTT, dithiothreitol; ECM, extracellular matrix; Edetic Acid - pharmacology; EDTA, ethylenediaminetetraacetic acid; Enzyme Activation - drug effects; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Gelatin - metabolism; Gelatinases; Glycoproteins - pharmacology; High molecular weight type IV collagenase; HPLC, high performance liquid chromatography; Humans; Hydrogen-Ion Concentration; Hydrolases; Liver Neoplasms - enzymology; Liver Neoplasms - secondary; Matrix Metalloproteinase 9; Metalloendopeptidases - chemistry; Metalloendopeptidases - metabolism; Metastatic carcinoma tissue; MMPs, matrix metalloproteinases; Molecular Weight; PAGE, polyacrylamide gel electrophoresis; Pepsin A - isolation & purification; Phenanthrolines - pharmacology; PMSF, phenylmethylsulfonyl fluoride; SDS, sodium dodecyl sulfate; Substrate Specificity; TIMP, tissue inhibitor of metalloproteinases; Tissue Inhibitor of Metalloproteinases; Trypsin - pharmacology; Type IV collagenase
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(93)80032-P

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.