EFFECT OF THIOCYANATE ON HUMAN SKIN COLLAGENASE ACTIVITY

<jats:title>ABSTRACT</jats:title><jats:p>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: <jats:list list-type="explicit-label"> <jats:list-item><jats:p> 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.</jats:p></jats:list-item> <jats:list-item><jats:p> Using the immunodiffusion technique, α<jats:sub>2</jats:sub>‐macroglobulin was detected up to the 4th day and α<jats:sub>1</jats:sub>‐antitrypsin until the 7th day.</jats:p></jats:list-item> <jats:list-item><jats:p> 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 α<jats:sub>1</jats:sub>‐antitrypsin, but no collagenase activity was detected in the area of α<jats:sub>2</jats:sub>‐macroglobulin.</jats:p></jats:list-item> <jats:list-item><jats:p> By treatment with NaSCN, the α<jats:sub>2</jats:sub>‐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 α<jats:sub>2</jats:sub>‐macroglobulin rather than α<jats:sub>1</jats:sub>‐antitrypsin.</jats:p></jats:list-item> </jats:list></jats:p>