抄録
Ac2-proteinase, one of proteinases of the venom of Agkistrodon acutus was purified by gel filtration on Sephadex G-75, followed by chromatographies on diethylaminoethyl (DEAE)-Sephadex A-50 and DE52 cellulose. By these procedures, 12.8 mg of purified preparation was obtained from 1g of crude venom. The purified preparation was homogeneous as judged by disc electrophoresis over polyacrylamide gel at pH 8.3 and isoelectric focusing. Ac2-proteinase also possessed both lethal and hemorrhagic activities. These and proteolytic activities were inhibited by ethylenediaminetetraacetic acid (EDTA), cysteine, or antiserum but not by soybean trypsin inhibitor or diisopropyl fluorophosphate. The molecular weight of Ac2-proteinase was estimated to be about 25000 by SDS-polyacrylamide gel electrophoresis, and the isoelectric point was found to be pH 4.9 by isoelectric focusing with carrier ampholyte. The minimum hemorrhagic dose, LD50, and proteolytic activities of the purified preparation were 0.431μg, 110μg/mouse, and 0.173 unit/mg, respectively. This protein did not contain any carbohydrates or nucleic acids.
