STUDIES ON THE ABILITY OF 65-KDA AND 92-KDA TUMOR-CELL GELATINASES TO DEGRADE TYPE-IV COLLAGEN

Two major gelatinolytic metalloproteinases (gelatinases) of 66 kDa and 92 kDa were purified from a tumor cell line. Analysis of collagen degradation showed that native full-length Engelbreth-Holm- Swarm (EHS) type IV collagen was not cleaved by the purified gelatinases under conditions where native pepsin-extracted human placental type IV and V collagen and heat-denatured collagens were markedly degraded. However, EHS type IV collagen degradation was noted at 37 OC, i.e. under conditions that would favor denaturation of the collagen molecule in solution. The pattern of degradation of human placental type IV and V collagen appeared similar for both gelatinases. Zymogram analysis of gelatinase activity in the absence of sodium dodecyl sulfate (SDS) (to eliminate possible SDS-mediated denaturation of type IV collagen) confirmed the inability of 65 and 92-kDa gelatinases to degrade native full-length EHS type IV collagen. Under the same conditions and in SDS-polyacrylamide gel electrophoresis zymograms the gelatinases degraded pepsin-predigested EHS type IV collagen and pepsin-extracted human placental type IV collagen. These data suggest that the 65- and 92-kDa tumor cell gelatinases are not true type IV collagenases. Their ability to degrade pepsin-solubilized, or denatured, type IV collagen suggests a specificity for telopeptide precleaved or conformationally altered forms of this molecule. Type IV collagen degradation