title: Extraction of collagenase from the 6000 times g sediment of uterine and skin tissues of mice. A comparative study.
abstract: An enzyme capable of digesting native collagen in solution at neutral pH was extracted from the 6 000 times g sediment of the involuting uterus of the mouse and of the back skins of mice and rats. The collagenase could be dissociated at cold-room temperature from the sediment in about equal amounts when neutral Tris buffer containing 1.0M NaCl or 5M urea was used for the extraction step. The enzyme has been concentrated by ammonium sulfate precipitation and the activity was measured by using [14C]collagen in solution at pH 7.5. Collagen breakdown products were identified by disc electrophoresis. The amount of enzyme extracted was a function of temperature and salt concentration. As 5M urea extracted collagenase from the sediment in a relatively short time, this method of extraction seems to be a useful tool for serial experiments in the study of collagenase activity in collagen-rich tissues.
mesh_terms: D000375:Aging; D000818:Animals; D003840:Deoxycholic Acid; D005260:Female; D051379:Mice; D003012:Microbial Collagenase / Q000302:isolation & purification*; D009928:Organ Specificity; D009994:Osmolar Concentration; D011092:Polyethylene Glycols; D011247:Pregnancy; D051381:Rats; D012867:Skin / Q000201:enzymology* / Q000254:growth & development; D013045:Species Specificity; D013696:Temperature; D013861:Thiocyanates; D014508:Urea; D014599:Uterus / Q000201:enzymology* / Q000254:growth & development
chemical_list: D013861:Thiocyanates; D003840:Deoxycholic Acid; D011092:Polyethylene Glycols; D014508:Urea; D003012:Microbial Collagenase