title: Isolation and characterization of synergistic hemorrhagins from the venom of the snake Bothrops asper.
abstract: Three hemorrhagic factors (BaH1, BH2 and BH3) were isolated from the venom of Bothrops asper by gel filtration on Sephacryl S-200, DEAE-Sepharose chromatography, metal chelate affinity chromatography and hydrophobic interaction chromatography. They contain 55% of the total hemorrhagic activity of the whole venom when they are mixed, but lose almost half of the activity if they are separated, indicating a synergism between the three. The main hemorrhagin is BaH1 (Bothrops asper hemorrhagin 1); the other two are weak hemorrhagins but contribute to the synergism. They are acidic proteins with a pI of 4.5, 5.2 and 5; their mol. wt is 64,000, 26,000 and 55,000 respectively. The minimal hemorrhagic dose (MHD) of BaH1, BH2 and BH3 is 0.18, 2 and 1.6 micrograms, with a specific activity 55, 5 and 6.25 higher than that of the whole venom. The hemorrhagic activity of all three factors was inhibited by EDTA and ortho-phenathroline, indicating that the hemorrhagic activity is metal dependent. Phosphoramidon, soybean trypsin inhibitor, PMSF, pepstatin and aprotinin did not affect the hemorrhagic activity of the isolated factors.
authors.affiliation: Department of Zoology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
authors.forename: G
authors.initials: G
authors.lastname: Borkow
authors.forename: J M
authors.initials: JM
authors.lastname: Gutiérrez
authors.forename: M
authors.initials: M
authors.lastname: Ovadia
mesh_terms: D010131:Aminosalicylic Acid; D000645:Ammonium Sulfate; D000818:Animals; D000918:Antibody Specificity; D000997:Antivenins / Q000276:immunology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003429:Cross Reactions; D003435:Crotalid Venoms / Q000737:chemistry* / Q000201:enzymology / Q000276:immunology; D004357:Drug Synergism; D010450:Endopeptidases / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification* / Q000633:toxicity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006470:Hemorrhage / Q000139:chemically induced; D006863:Hydrogen-Ion Concentration; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007525:Isoelectric Focusing; D051379:Mice; D008970:Molecular Weight
chemical_list: D000997:Antivenins; D003435:Crotalid Venoms; D004791:Enzyme Inhibitors; D010131:Aminosalicylic Acid; D010450:Endopeptidases; C041605:hemorrhagic proteinase IV; D000645:Ammonium Sulfate