EFFECT OF THIOCYANATE ON HUMAN SKIN COLLAGENASE ACTIVITY

Using tissue culture, human skin collagenase activity and its relationship to serum collagenase inhibitors and the effect of thiocyanate was studied. The following was found: 1) Collagenase activity did not become apparent in the culture medium before 48 hours; it reached a peak on the 4th day, gradually decreased, and then became elevated again on the 7th day of culture. 2) Using the immunodiffusion technique, α2-macroglobulin was detected up to the 4th day and α1-antitrypsin until the 7th day. 3) On the 2nd day of culture, two separate peaks of collagenase activity were evident by gel filtration on a Sephadex G-200 column. One peak was eluted in the area of α1-antitrypsin, but no collagenase activity was detected in the area of α2-macroglobulin. 4) By treatment with NaSCN, the α2-macroglobulin was dissociated from the enzyme-serum collagenase inhibitor complex, and skin collagenase was partially purified as a single peak of activity. From these results, the main serum constituent of collagenase inhibitors is considered to be α2-macroglobulin rather than α1-antitrypsin.