CHARACTERISTICS OF 92 KDA TYPE-IV COLLAGENASE GELATINASE PRODUCED BY GRANULOCYTIC-LEUKEMIA CELLS - STRUCTURE, EXPRESSION OF CDNA IN ESCHERICHIA-COLI AND ENZYMATIC-PROPERTIES/

Human neutrophils can be triggered to release the collagenolytic metal loenzymes, interstitial collagenase and 92 kDa type IV collagenase/gel atinase. We have isolated and sequenced a 2.3 kb cDNA from a chronic g ranulocytic leukemia cDNA library that encodes for human neutrophil ty pe IV collagenase. With the exception of one amino-acid substitution a t position 280 (Arg --> Gln), the deduced amino-acid sequences of neut rophil gelatinase are identical to the amino-acid sequences of the enz yme isolated from fibrosarcoma cells. Expression of the cDNA in E. col i yielded a 72 kDa protein having a gelatinolytic activity on zymogram gel. The recombinant enzyme was activated with APMA and trypsin. The activation was accompanied by a reduction in molecular weight of appro ximate to 10 kDa; such a reduction is characteristic of matrix metallo proteinases. The recombinant gelatinase cleaved native type V and XI c ollagens. Native type I collagen was not a substrate for the enzyme. T hese data suggest that native and recombinant 92 kDa type IV collagena se produced in E. coli have similar biochemical properties. The succes sful expression of the collagenase in a prokaryotic system will greatl y facilitate the structure-function characterization of the enzyme and allow a more precise analysis of its physiological and pathological r oles.