Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and dis...Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and disintegrin domains. However, no hemorrhagic and fibrinogenolytic activities were detected for Jerdonitin. We thought that organic buffer of high performance liquid charamatography （HPLC） might affect its enzymatic activity. In this study, we purified Jerdonitin by another procedure excluding the HPLC. It was homogenous as judged by SDS-PAGE and had an apparent molecular weight of 36 kDa under non-reducing conditions and 38 kDa under reducing conditions, respectively. Like other typical metalloproteinases, Jerdonitin preferentially degraded alpha-chain of human fibrinogen and this fibrinogenolytic activity was completely inhibited by EDTA, but not by PMSF. It was interesting that Jerdonitin did not induce hemorrhage after intradermal injection in mice.展开更多
Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated fr...Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated from the venom of Chinese Trimeresurus mucrosqumatus by a combination of ion-exchange chromatography on DEAE-Sephadex A-50, CM-Sepharose Cl-6B and gel filtration on Sephadex G-100.Results: The purified protein named TM-AEH,a glycoprotein with carbohydrate content of 0.5% neutral hexose and 0.75% sialic acid,a relative molecular mass of 29.0 kDa,and an isoelectric point(pI) of 5.2. It shares with an extinction coefficient(E 0.1%/cm) of 1.332 at 280 nm,consisted of 225 amino acid residues,and migrated as a band under reduced or non-reduced condition in basic PAGE.TM-AEH was a highly thermostable protein and was stable to pH changes between 5 and 9.The optimum temperature and optimum pH were 55℃ and 8.4 for its catalytic activity respectively,which was inhibited by Fe 3+ and Cu 2+.Conclusion:This protein can exhibit higher BAEE-hydrolysing activity and fibrinogenolytic activity as compared to that of whole venom.展开更多
文摘Previously, we have purified Jerdonitin from Trimeresurusjerdonii venom. Compared with other P-Ⅱ class snake venom metalloproteinases （SVMPs）, Jerdonitin has a primary structure comprising metalloproteinase and disintegrin domains. However, no hemorrhagic and fibrinogenolytic activities were detected for Jerdonitin. We thought that organic buffer of high performance liquid charamatography （HPLC） might affect its enzymatic activity. In this study, we purified Jerdonitin by another procedure excluding the HPLC. It was homogenous as judged by SDS-PAGE and had an apparent molecular weight of 36 kDa under non-reducing conditions and 38 kDa under reducing conditions, respectively. Like other typical metalloproteinases, Jerdonitin preferentially degraded alpha-chain of human fibrinogen and this fibrinogenolytic activity was completely inhibited by EDTA, but not by PMSF. It was interesting that Jerdonitin did not induce hemorrhage after intradermal injection in mice.
文摘Objective: To study the physical and chemical properties of an arginine ester hydrolase from the venom of Trimeresurus mucrosqumatus in Hunan province of China.Methods:The arginine ester hydrolase(AEH) was isolated from the venom of Chinese Trimeresurus mucrosqumatus by a combination of ion-exchange chromatography on DEAE-Sephadex A-50, CM-Sepharose Cl-6B and gel filtration on Sephadex G-100.Results: The purified protein named TM-AEH,a glycoprotein with carbohydrate content of 0.5% neutral hexose and 0.75% sialic acid,a relative molecular mass of 29.0 kDa,and an isoelectric point(pI) of 5.2. It shares with an extinction coefficient(E 0.1%/cm) of 1.332 at 280 nm,consisted of 225 amino acid residues,and migrated as a band under reduced or non-reduced condition in basic PAGE.TM-AEH was a highly thermostable protein and was stable to pH changes between 5 and 9.The optimum temperature and optimum pH were 55℃ and 8.4 for its catalytic activity respectively,which was inhibited by Fe 3+ and Cu 2+.Conclusion:This protein can exhibit higher BAEE-hydrolysing activity and fibrinogenolytic activity as compared to that of whole venom.
