The recombinant catalytic domain of human neutrophil collagenase lacks type I collagen substrate specificity.

The coding region for human neutrophil short form procollagenase lacking the hemopexin like domain coding region was amplified by polymerase chain reaction. Recombinant short form procollagenase was expressed in E. coli and purified in a three step procedure. Renaturation of this proenzyme was carried out by an effective new method using Q-Sepharose chromatography. Treatment of short form procollagenase with mercurials resulted in active short form collagenase M(r) 21,000 and an intermediate product of M(r) 23,000. These two products were separated by hydroxamate affinity chromatography. The active, short form collagenase M(r) 21,000 is stable. Despite full proteolytic activity, it lacks type I collagen substrate specificity and forms the basis for crystallisation experiments.