A PURIFICATION AND CHARACTERIZATION OF HIGH MOLECULAR WEIGHT HEMORRHAGIN PRESENT IN THE SNAKE VENOM Bothrops pictus

Abstract

A hemorrhagin with metalloprotease activity was purified from the venom of Bothrops pictus
snake using Sephadex G-75 molecular gel filtration and DEAE A-50 ionic exchange column.
Thus a homogeneus protein entity was obtained with 62 kDa under non reducting conditions.
Hemorrhagin is a acid protein, attack both casein and collagen being 0,226 μg as a DHM.
Chelating agent such as EDTA as well as 2 ß-mercaptoetanol and DTT produced strong
inhibition both caseinolitic and hemorrhagic activities. Optimus pH was 7,5 and heating
treatment reduced both activities, At 55 °C recovered activity on casein was 30,4%. On the
other hand hemorrhagin is an antigenic entity showed on double immunodiffusion test and
was neutralizated fulling with 0,5, 1 and 2 doses of antivenom.