Extraction of collagenase from the involuting rat uterus

摘要: Abstract Collagenase (EC 3.4.24.3) activity can be measured directly in homogenates of the involuting rat uterus. Latent forms collagenase are activated by a brief exposure to trypsin; trypsin is then blocked with soybean inhibitor. Homogenizing conditions have been developed that permit 90–95% recovery total active and latent 6000 × g pellet, where it presumably bound its collagen substrate. This insoluble extracted heating 60°C for 4 main 0.04 M Tris · HCl buffer, pH 7.5, containing 0.1 CaCl2. Methods presented estimation extracts; this approximates 65–70% total. Small amounts also from liver kidney. extraction procedure should use purifying without culturing enzyme-producing tissue direct assay activity. The uterus has proven characteristic pattern breakdown products on disc electrophoresis split tropocollagen at interband 41 as shown electron microscopy reconstituted fragments. inhibited EDTA, inhibition not reversed calcium or zinc ions.