백편두 (Dolichos lablab)에 존재하는 serine protease inhibitor의 분리, 정제 및 그 생화학적 특성에 관한 연구 Purification and characterization of three kinds of serine protease inhibitors from dolichos lablab seeds원문보기
Three kinds of serine protease inhibitors (named TI-1, TI-2, and TI-3) were purified to homogeneity and characterized their biochemical properties from Dolichos lablab seeds. TI-1 showed the strongest inhibitory effects against chymotrypsin. TI-2 strongly inhibited the activity of porcine pancreatic...
Three kinds of serine protease inhibitors (named TI-1, TI-2, and TI-3) were purified to homogeneity and characterized their biochemical properties from Dolichos lablab seeds. TI-1 showed the strongest inhibitory effects against chymotrypsin. TI-2 strongly inhibited the activity of porcine pancreatic elastase among known serine proteases. TI-3 showed the inhibitory activities against trypsin and plasmin. On SDS-PAGE under reducing conditions, TI-1, TI-2, and TI-3 gave a single bands on 15.9, 12.1, and 14.6 kDa, respectively. The purified TIs did not exhibit the inhibitory effects on α-thrombin and plasma kallikrein. To determine their structures, we determined their partial amino acid sequence analysis. TI-1, TI-2, and TI-3 showed a homologies with Soybean Bowman-Birk inhibitors (BBI). Interestingly, TI-3 showed the same gel mobility with that of Soybean Kunitz type inhibitors on SDS-PAGE under non-reducing conditions. These results suggested that TI-3 might be existed as intra or inter-molecular dimeric structure. To estimate the biological functions of the purified inhibiotrs, we used two animal model system; 1) pseudomonal elastase-induced septic shock model, 2) trypsin-induced lethal shock model. For biological experiments, we used TI mixture, showing the same inhibitory effects with the purified TIs. Pretreatment of 33 mg/kg of TI mixture, we observed the inhibitory effects on pseudomonal elastase-induced hypotension on guinea pig. Also, bradykinin and kallikrein in serum were not increased by the pretreatment of TI mixture. Moreover, TM (330 mg/kg) showed a predominent inhibitory effect on trypsin-induced lethal shock in mice. These results suggested that the purifed inhibitors might be able to use for treating bacterial protease-induced septic shock disease.
Three kinds of serine protease inhibitors (named TI-1, TI-2, and TI-3) were purified to homogeneity and characterized their biochemical properties from Dolichos lablab seeds. TI-1 showed the strongest inhibitory effects against chymotrypsin. TI-2 strongly inhibited the activity of porcine pancreatic elastase among known serine proteases. TI-3 showed the inhibitory activities against trypsin and plasmin. On SDS-PAGE under reducing conditions, TI-1, TI-2, and TI-3 gave a single bands on 15.9, 12.1, and 14.6 kDa, respectively. The purified TIs did not exhibit the inhibitory effects on α-thrombin and plasma kallikrein. To determine their structures, we determined their partial amino acid sequence analysis. TI-1, TI-2, and TI-3 showed a homologies with Soybean Bowman-Birk inhibitors (BBI). Interestingly, TI-3 showed the same gel mobility with that of Soybean Kunitz type inhibitors on SDS-PAGE under non-reducing conditions. These results suggested that TI-3 might be existed as intra or inter-molecular dimeric structure. To estimate the biological functions of the purified inhibiotrs, we used two animal model system; 1) pseudomonal elastase-induced septic shock model, 2) trypsin-induced lethal shock model. For biological experiments, we used TI mixture, showing the same inhibitory effects with the purified TIs. Pretreatment of 33 mg/kg of TI mixture, we observed the inhibitory effects on pseudomonal elastase-induced hypotension on guinea pig. Also, bradykinin and kallikrein in serum were not increased by the pretreatment of TI mixture. Moreover, TM (330 mg/kg) showed a predominent inhibitory effect on trypsin-induced lethal shock in mice. These results suggested that the purifed inhibitors might be able to use for treating bacterial protease-induced septic shock disease.
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