![]() | Only 14 pages are availabe for public view |
Abstract In the present years , it has been noticed in the international scientific literatures a rapid increase in the studies of animal venoms and their constituents not only as research tools for the neurologists, hematologists, physiologists and pharmacologists but also mainly in the pharmaceutical industry , for the development of potential veterinarian and human drugs. Consequently, this submitted work was conducted to look for the polypeptides responsible for the BK potentiating activity in the venom of the scorpion Buthus occitanus. The scorpions were milked by electric stimulation (6volts) at the articular membrane of the telson . from 1500 scorpions, about one gram of.dry venom was obtained within 4 months . The crude venom was dissoluted in distilled water and then centrifuged. The supernatant was collected and dialyzed with spectrapore 3 membrane (cut off M.W. 3500 ) for three days at 4 °C. The dialysate and dialyzed venom were lyophilized and then quantitated by amino acid analysis after acid hydrolysis . The dialysate fraction was not toxic for mice at doses up to 100 lA-g/mouse via intracerebroventricular injection. The potentiation of BK on the isolated guinea pig ileum and the rat uterus were the biological models used to follow the BK potentiating .activity in each step of purification.BK potentiating activity was found in dialysate separated from the bulk of the venom protein.Six chromatographic fractions (A to F) from dialysate were isolated by RP-HPLC. Each fraction was collected separatelly, lyophilized and then quantitated by amino acid composition after acid hydrolysis . Fractions C and F were found as BK potentiators.With system used, fraction F was more hydrophobic as well as more potent than fraction C. Further RP-HPLC purification of these fractions was performed.Ten peptides were obtained homogeneous under the conditions used. One peptide (C2) in fraction C and seven peptides (Fl, F3, F4-2, F5-2, F6, F7 and Fg) in fraction F exhibited BK potentiating activity. All the studied fractions and peptides were tested for BK. |