Identification and partial characterisation of hyaluronidases in Lonomia obliqua venom.
By studying Lonomia obliqua (caterpillar) venom we were able to detect a lytic activity on purified hyaluronic acid. The venom hydrolyses purified chondroitin sulphate, but was unable to degrade either heparan sulphate or dermatan sulphate. Moreover, through purified hyaluronic acid-degrading kinetic assays, we observed that this lytic activity was caused by a hydrolase rather than lyase enzyme. In addition, by using the Reissig colorimetric reaction, we detected this hyaluronic acid hydrolase action as a beta-endohexosaminidase enzyme originating terminal N-acetylglucosamine residues rather than beta-endoglucuronidase, which may originate glucuronic acid residues. Zymogram analysis of the venom detected 49 and 53 kDa molecules with hyaluronic acid lytic activity. An examination of these hyaluronic acid degrading activities as a function of pH showed that these hydrolases had no apparent activities at a pH below 5.0 and higher than 8.0 and displayed their optimal activities at pH ranging from 6.0 to 7.0. Finally, through a fluorescence reaction to hyaluronic acid and confocal microscopy, we confirmed this cleaving action upon hyaluronic acid organised on the extracellular matrix of the dermis of rabbit. The data provide experimental evidence of the presence of hyaluronidases in the L. obliqua venom, probably involved in the harmful effects of the venom.
da C B Gouveia
Ana Isabel
AI
Department of Cell Biology, Federal University of Paraná, Jardim das Américas, 81531-990 Curitiba, Paraná, Brazil.
da Silveira
Rafael B
RB
Nader
Helena B
HB
Dietrich
Carl P
CP
Gremski
Waldemiro
W
Veiga
Silvio S
SS
D000818:Animals; D001180:Arthropod Venoms / Q000737:chemistry / Q000201:enzymology*; D002809:Chondroitin Sulfates / Q000378:metabolism; D003124:Colorimetry; D020405:Dermis / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism; D005453:Fluorescence; D006820:Hyaluronic Acid / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D018613:Microscopy, Confocal; D009036:Moths / Q000737:chemistry* / Q000201:enzymology; D011817:Rabbits
D001180:Arthropod Venoms; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates