Extractable Collagenase and Carcino Genesis of the Mouse Skin

Abstract

Collagenolytic activity has been demonstrated in the early phase of chemical carcinogenesis of mouse skin following 3-methylcholanthrene application dropwise in acetone or painted on the skin in benzene. In addition very high levels of collagenase could be detected in mouse skin papillomas and carcinomas. In all the tissues investigated, collagenase activity was extracted from the 6000 × g sediment of tissue homogenates with 5 M urea in 50 mM Tris-HCl buffer, pH 7.5. After dialyzing the extract, the enzyme was precipitated with ammonium sulfate and the activity determined against ¹⁴C-collagen substrate in solution. This procedure was found suitable for the detection and estimation of collagenase activity in skin tissues with high turnover of collagen and thus offers an attractive alternative to tissue culture methods.