2. Academic Validation
  3. Spectrophotometric assay for vertebrate collagenase

Spectrophotometric assay for vertebrate collagenase

  • Anal Biochem. 1985 Jun;147(2):437-40. doi: 10.1016/0003-2697(85)90294-5.
H Weingarten,  J Feder
PMID: 2990254 DOI: 10.1016/0003-2697(85)90294-5
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.

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