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An improved assay of mammalian collagenase activity, and its use to determine hepatic extracellular matrix susceptibility to degradation.

1982; American Association for Clinical Chemistry; Volume: 28; Issue: 10 Linguagem: Inglês

10.1093/clinchem/28.10.2134

1530-8561

William J. Lindblad, George C. Fuller,

Bariatric Surgery and Outcomes

This rapid, sensitive method of determining collagenase (EC 3.4.24.7) activity incorporates several advantages of previous methods. Soluble [14C]acetylated collagen is prepared as the enzyme substrate and collagen-cleavage products are separated from noncleaved collagen by precipitation with dioxane/methanol. The assay is more reproducible than previous methods and has a lower detection limit, 15 mU of enzyme activity. We used the method in a competitive substrate assay with isolated extracellular hepatic matrix from cirrhotic and normal rat liver. Purified collagenase was consistently bound to normal rat matrix to a greater extent than to cirrhotic matrix, suggesting that in hepatic fibrosis the extracellular matrix is not as susceptible to collagenase degradation as that in normal liver.