title: An improved assay of mammalian collagenase activity, and its use to determine hepatic extracellular matrix susceptibility to degradation.
abstract: 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.
mesh_terms: D000818:Animals; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008103:Liver Cirrhosis / Q000378:metabolism; D003012:Microbial Collagenase / Q000032:analysis*; D051381:Rats; D013379:Substrate Specificity
chemical_list: D003094:Collagen; D003012:Microbial Collagenase