Purification of Haemolytic/Necrotic Protein from the Venom of Deathful Hemiscorpius Lepturus Scorpion in Khuzestan Province

Scorpion envenomation is a public health problem in Iran. Scorpions are the second accident after snake envenomation in causing human fatalities. Hemiscorpius lepturus is the most medically important scorpion in Iran with 95% mortality. The venom from H. lepturus is primarily a cytotoxic agent and has hemolytic dermonecrotic, nephrotoxic, and to some extent, hepatotoxic activities. H. lepturus venom collected from Khuzestan province. Reverse phase HPLC (RP-HPLC) was performed to analyze proteins and peptides in crude venom. The collected fractions were dried and re-solubilized in dH2O. The haemolytic test done on obtained RP-HPLC fractions and their molecular weights were determined. The positive fractions in haemolytic test were subjected to dermonecrotic test in Rabbit. The number of 103 fractions collected from C18 RP-HPLC column in the crude venom of H. lepturus. Four fractions with severe hemolytic activities were selected. The SDS-PAGE showed 32-32kDa proteins. The amount of 5 and 20 μg of hemolytic positive fractions were subjected to the dermonecrotic test on rabbit skin and the severity of lesions were studied. These fractions showed a different necrosis level in the skin that the highest activity belonged to the fraction in 157.55 min elution time. The analysis of the protein contents of the venom of Hemiscorpius lepturus showed that the major toxins are related to the proteins with haemolytic/dermonecrotic activities. The severity of four identified proteins is due to the different isoforms of a major protein.