In Brazil, accidents with scorpion stings have been a serious public health problem, and Tityus serrulatus (Ts) is primarily responsible for severe accidents. Epitopes common to both isoforms included active site residues. More peptides were recognized by anti-hyaluronidase serum in TsHyal-1 than in TsHyal-2. We used the SPOT method to map TsHyal-1 and TsHyal-2 epitopes. Addition of native Ts hyaluronidase (0.418 µg) to pre-neutralized Ts venom (13.2 µg venom+0.94 µl anti-hyaluronidase serum) reversed mouse survival. Inhibition of death was also achieved by using the hyaluronidase pharmacological inhibitor aristolochic acid. In vivo neutralization assays showed that 121.6 µl of anti-hyaluronidase serum inhibited mouse death 100%, whereas 60.8 µl and 15.2 µl of serum delayed mouse death. As little as 0.94 µl of anti-hyaluronidase serum neutralized 1 LD 50 (13.2 µg) of Ts venom hyaluronidase activity in vitro. ![]() Purified native Ts hyaluronidase was used to produce anti-hyaluronidase serum in rabbits. We performed gel filtration followed by reversed-phase chromatography to purify native hyaluronidase from Ts venom. ![]() The isoforms are 83% identical, and alignment of their predicted amino acid sequences with other hyaluronidases showed conserved residues between evolutionarily distant organisms. ![]() We examined clones from a cDNA library of the Ts venom gland and described two novel isoforms of hyaluronidase, TsHyal-1 and TsHyal-2.
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