ISOLATION OF CROTAMINE FROM VENOM OF ARGENTINIAN RATTLESNAKE

FUSCO, L; LEIVA, L; ECHEVERRÍA S.M.; BUSTILLO, S.; GAUNA M.C

Buenos Aires

Jornada; XVIII Jornadas Anuales Multidisciplinarias de Biología; 2016

Sociedad Argentina de Biología

Crotamine is a small cationic peptide originally found in the venom of the South American rattlesnake Crotalus durissus terrificus. It is a 5-kDa mio-neurotoxin that has been shown to possess analgesic effects. At final concentration ranging from 0,1 to 10 M, it was not cytotoxic to normal cells of different types (e.g., muscle cells, human endothelial cells), even after 72 h of exposure, according to works of brazilian authors. Considering regional variation not only between species, but also within a single species, it is our interest to isolate and characterize crotamine from specimens living in the Argentine northeast, which has not been studied yet. In this work we purified crotamine by size-exclusion chromatography and ultrafiltration. When injected i.p. (8,75- 280 ng/Kg body mass) in adult male CF1 mice,it induced a time-dose dependent high analgesic effect by the acetic acid-induced writhing method, where 35 ng/Kg was the minor dose that showed effect. By in vitro assays, at final concentration of 0,53 M, the crotamine isolated from argentinean rattlesnakes, exhibited toxic activity on C2C12 cells, in contrast to the behavior of that obtained from brazilian snake venoms. These preliminary findings highlight differences between crotamines purified from species from distinct geographic areas. Further study is required to elucidate the structure and the properties of the small protein here isolated.