Hyaluronidase polymorphism detected by polyacrylamide gel electrophoresis. Application to hyaluronidases from bacteria, slime molds, bee and snake venoms, bovine testes, rat liver lysosomes, and human serum.
A gel electrophoretic technique which allows detection of hyaluronidase activity in the gel has been devised. The principle is that the high-molecular-weight substrate, hyaluronic acid, is included in the gel, where it cannot move in the electrical field. After the run, the gel is incubated under conditions allowing the enzyme to degrade the substrate. Upon staining with "Stains-all" dye (Eastman Kodak Co., 2718), zones of hyaluronidase activity appear as pink bands in a blue background. The sensitivity limit is less than 3 fkat equivalent to 2.2 NF mU. The method is applicable to all types of hyaluronidases and chondroitinase ABC. It enabled to be shown that some hyaluronidases are polymorphic. This technique also made it possible to detect easily hyaluronidase activity in normal human serum. This analytical method represents a convenient step in the purification of hyaluronidase.
D000818:Animals; D001419:Bacteria / Q000201:enzymology; D001514:Bee Venoms / Q000032:analysis; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000097:blood / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D009235:Myxomycetes / Q000201:enzymology; D051381:Rats; D011919:Rats, Inbred Strains; D012910:Snake Venoms / Q000032:analysis; D013045:Species Specificity; D013194:Staining and Labeling; D013737:Testis / Q000201:enzymology
D001514:Bee Venoms; D012910:Snake Venoms; D006821:Hyaluronoglucosaminidase