Purification and renal effects of phospholipase A2 isolated from Bothrops insularis venom

Bothrops insularis venom contains a variety of substances presumably responsible for several pharmacological effects. We investigated the biochemical and biological effects of phospholipase A2 protein isolated from B. insularis venom and the chromatographic profile showed 7 main fractions and the main phospholipase A2 (PLA2) enzymatic activity was detected in fractions IV and V. Fraction IV was submitted to a new chromatographic procedure on ion exchange chromatography, which allowed the elution of 5 main fractions designated as IV-1 to IV-5, from which IV-4 constituted the main fraction. The molecular homogeneity of this fraction was characterized by high-performance liquid chromatography (HPLC) and demonstrated by mass spectrometry (MS), which showed a molecular mass of 13984.20 Da; its N-terminal sequence presented a high amino acid identity (up to 95%) with the PLA2 of Bothrops jararaca and Bothrops asper. Phospholipase A2 isolated from B. insularis (Bi PLA2 ) venom (10 μg/mL) was also studied as to its effect on the renal function of isolated perfused kidneys of Wistar rats (n=6). Bi PLA2 increased perfusion pressure (PP), renal vascular resistance (RVR), urinary flow (UF) and glomerular filtration rate (GFR). Sodium (%TNa+) and chloride tubular reabsorption (%TCl−) decreased at 120 min, without alteration in potassium transport. In conclusion, PLA2 isolated from B. insularis venom promoted renal alterations in the isolated perfused rat kidney.