title: Spectrophotometric assay for vertebrate collagenase.
abstract: Collagenase from normal human skin fibroblasts was found to catalyze the hydrolysis of esters and thio esters. This observation led to the development of a rapid, sensitive, continuous spectrophotometric assay for vertebrate collagenase using the thio peptolide Ac-ProLeuGly-S-LeuLeuGly-OC2H5 as substrate in the presence of 4,4'-dithiodipyridine or Ellman's Reagent. A Km of 0.004 M and a kcat of 370,000 h-1 were determined for the thio peptolide-enzyme reaction. The method is able to detect collagenase at concentrations as low as 2 ng/ml.
authors.forename: H
authors.initials: H
authors.lastname: Weingarten
authors.forename: J
authors.initials: J
authors.lastname: Feder
mesh_terms: D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D003012:Microbial Collagenase / Q000032:analysis*; D012867:Skin / Q000201:enzymology; D013053:Spectrophotometry
chemical_list: D003012:Microbial Collagenase