A vertebrate interstitial collagenase inhibitor from bovine scapular cartilage: purification and characterization.

A collagenase inhibitor was purified from bovine cartilage by a combination of gel filtration, ion exchange, concanavalin A-Sepharose affinity chromatography, and elution from preparative sodium dodecyl sulfate-polyacrylamide gels. The inhibitor was purified 370-fold and migrated as a single polypeptide with an Mr of 19,000 on SDS-polyacrylamide gels. It stained positively for carbohydrate with periodic acid-Schiff's reagent and bound to lectins, indicating that it is a glycoprotein. The inhibitory activity was stable to heating up to 60 degrees C and between pH 4 and 10. The inhibition of collagenase by the cartilage inhibitor could not be reversed by trypsin or mersalyl. The inhibitory activity did not require the presence of free sulfhydryl groups, and it could be removed from the cartilage extract by incubating with native collagen, suggesting that the inhibitor binds to collagen. The cartilage inhibitor was effective against human and mouse interstitial collagenases, but it did not inhibit trypsin or bacterial collagenase.