The collagenase of Entamoeba histolytica.
The present work was designed to investigate the capacity of trophozoites of Entamoeba histolytica to adhere to and digest human collagen types I and III in vitro. The time-course of binding of ameba to both human collagen types I and III was similar. However, the kinetics of detachment were different for each collagen type. Trophozoites of E. histolytica cultured on heat-reconstituted type I collagen gels produced a well-defined area of lysis. Quantitative studies using 14C-labeled collagen revealed that after 24 h of incubation, Entamoeba digested three and a half times more type I than type III collagen, thus suggesting the presence of a collagenase with higher specificity for type I collagen. This activity was optimum with trophozoites harvested after 42 h in culture (1.5 X 10(5) trophozoites/ml). The digestion of type I collagen was a function of the number of trophozoites, and was inhibited by EDTA, L-cysteine, and serum, but not by soybean trypsin inhibitor, phenylmethanesulfonyl fluoride, or N-ethylmaleimide (NEM). Electrophoretic analysis of the type I collagen fragments revealed three main classes of polypeptides of 75,000, 50,000, and 25,000 daltons. Subsequent proteolysis of these collagen fragments was probably carried out by other proteases derived from trophozoites. This activity was inhibited with 10 mM NEM. Collagenase activity appeared to be located at the plasma membrane and direct contact of the ameba with the substrate is required for collagen digestion. The results suggest that collagenase activity of E. histolytica may play an important role in tissue invasion.
D000268:Adhesiveness; D000818:Animals; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000254:growth & development / Q000469:parasitology; D005615:Freezing; D007700:Kinetics; D003012:Microbial Collagenase / Q000378:metabolism*; D014461:Ultracentrifugation
D003470:Culture Media; D003094:Collagen; D003012:Microbial Collagenase