Spectrophotometric assay for vertebrate collagenase

✍ Scribed by Harold Weingarten; Joseph Feder

• Book ID: 102629786
• Publisher: Elsevier Science
• Year: 1985
• Tongue: English
• Weight: 252 KB
• Volume: 147
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(85)90294-5

✦ Synopsis

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-OCzHs as substrate in the presence of 4,4'dithiodipyridine or Ellman's Reagent. A Km of 0.004 M and a k,,, of 370,000 h-' were determined for the thio peptohde-enzyme reaction. The method is able to detect collagenase at concentrations as low as 2 rig/ml. o 1985 Academic Press. Inc.