Partial purification and characterization of a neutral protease which cleaves the N-terminal propeptides from procollagen.
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A rapid assay procedure was developed for cleavage of the N-terminal propeptides of procollagen. With the assay a neutral procollagen N-protease was purified about 300-fold from chick embryo tendon extract. The enzyme had an apparent molecular weight of 260 000 and a pH optimum of 7.4. Ca2+ was required for enzymic activity but this requirement was partially replaced by Mg2+ or Mn2+. The enzyme was bound to concanavalin A-agarose and therefore was presumably a glycoprotein. The N-propeptides released from type I procollagen were of about 23 000 and 11 000 daltons as estimated by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. The partially purified enzyme was also found to cleave type II procollagen and the N-propeptide obtained was about 18 000 daltons. Heat denaturation of either type I or type II procollagen decreased the rate at which the proteins were cleaved by the N-protease.