Evaluation of moniotoxic activity of bothropstoxin-1 irradiated with sup(60)Co gamma rays

Abstract

Ionizing radiation has been successfully employed to modify the immunological properties of biomolecules and has been proven to be a powerful tool to attenuate snake venoms toxicity without affecting their immunogenic properties. Snake venoms and their isolated toxins showed structural modifications after gamma radiation process, in aqueous solution, mainly by water radiolysis sub products. Free radical scavengers, such as NaNO3 and t-butanol, present selective effects on specific radical from water radiolysis. The NaNO3 has affinity by aqueous electron, while the t-butanol has affinity by hydroxyl radical. At the present work, we have investigated the miotoxic activity of bothropstoxin-1 (BTHX-1), a K49 phospholipase, present in Bothrops jararacussu crude venom, before and after irradiation process, with or without scavenger substances presence. BTHX-1 was irradiated with 2kGy of 60Co gamma rays, in aqueous solution and in the presence of oxygen. BALB/c mice were inoculated with either native or irradiated toxin, with or without scavenger substances. After 3 hours, blood samples were collected and the miotoxic activity was evaluated by LDH (lactate dehydrogenase) release. The muscular tissue damage was directly related to the LDH amounts released. Irradiated bothropstoxin-1, with or without NaNO3 substance, caused less damage than their native counterpart. But irradiated toxin, in the presence of t-butanol, was so miotoxic as the native BTHX-1. These results indicate that irradiation of toxic proteins can promote significant modifications on their structures, but still retaining many of the original biological properties of their native counterparts. Additionally, some scavengers substances can change these gamma radiation effects.