[Characterization of lysosomal-type hyaluronidase in the culture medium of 2 cell lines derived from human hepatomas].
A sensitive assay for hyaluronidase was developed using as a substrate, hyaluronic acid insolubilized on polystyrene microtest plates. Hyaluronic acid was measured exploiting the fact that it can bind immune complexes made up with hyaluronectin and alkaline phosphatase-conjugated anti-hyaluronectin antibodies. Hyaluronidase was detected in both cell line culture media. Optimum pH was between 3.25 and 3.75. Sodium chloride dependence was absolute, and the optimum concentration of sodium chloride was between 0.2 and 0.3 M. The activity was not affected by dialysis, and was suppressed by a 5 minute heating at 50 degrees C or by protease treatment. The molecular weight was 68 K as determined by gel permeation chromatography. The results are close to those reported for human lysosomal hyaluronidase.
D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D002460:Cell Line; D003470:Culture Media; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008113:Liver Neoplasms / Q000201:enzymology*; D008247:Lysosomes / Q000201:enzymology*
D003470:Culture Media; D006821:Hyaluronoglucosaminidase
Caractérisation d'une hyaluronidase de type lysosomal dans le milieu de culture de deux lignées cellulaires dérivées d'hépatomes humains.