Assay of collagenase activity by a rapid, sensitive, and specific method.
A new method for the assay of collagenase activity has been developed, whereby the collagen cleavage products, after initial collagenase digestion, are degraded further by a mixture of trypsin and alpha-chymotrypsin. The degradation products are soluble in TCA and can be conveniently separated from the remaining uncleaved collagen substrate by rapid filtration. The enzyme assay is shown to be reproducible and sensitive, and it lends itself to a convenient and rapid determination of collagenase activity in relatively large numbers of samples. The applicability of this method is demonstrated by the detection of increased collagenase activity in skin fibroblast cultures derived from a patient with recessive dystrophic epidermolysis bullosa.
D001419:Bacteria / Q000201:enzymology; D002918:Chymotrypsin; D004820:Epidermolysis Bullosa / Q000201:enzymology; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D008722:Methods; D003012:Microbial Collagenase / Q000032:analysis*; D009504:Neutrophils / Q000201:enzymology*; D012867:Skin / Q000201:enzymology*; D012995:Solubility; D013696:Temperature; D014238:Trichloroacetic Acid; D014357:Trypsin
D014238:Trichloroacetic Acid; D002918:Chymotrypsin; D014357:Trypsin; D003012:Microbial Collagenase