By successive chromatography on CM-Sepharose CL-6B,Sephadex G-100,Heparin-Sepharose CL-6B, a phosphodiesterase (PDE, EC3.1.4.1) was isolated and purified from Guangxi cobra venom. The final preparation was 89 fold pur...By successive chromatography on CM-Sepharose CL-6B,Sephadex G-100,Heparin-Sepharose CL-6B, a phosphodiesterase (PDE, EC3.1.4.1) was isolated and purified from Guangxi cobra venom. The final preparation was 89 fold pure and had high specific activity. The enzyme showed a single band on SDS-PAGE and IEF. The PDE showed a molecular weight of 110 kD and pI of 7.4. The enzyme consists of 840 amino acid residues and has only a methionine as N-terminal amino acid residues. It was rich in asparagine, leucine and glutamine and poor in sulphur-containing amino acid. It contains 5.38% neutral sugars. It has no acute toxicity and has analgesia effect.展开更多
目的从短尾蝮蛇毒中分离纯化一种抗凝蛋白,并对其生化性质进行研究。方法利用阳、阴离子交换、凝胶过滤的方法分离纯化这种抗凝蛋白,用活化部分凝血活酶时间(APTT)测定其抗凝活性,用SDS-PAGE测定其蛋白相对分子量,用等电聚焦法测定蛋白...目的从短尾蝮蛇毒中分离纯化一种抗凝蛋白,并对其生化性质进行研究。方法利用阳、阴离子交换、凝胶过滤的方法分离纯化这种抗凝蛋白,用活化部分凝血活酶时间(APTT)测定其抗凝活性,用SDS-PAGE测定其蛋白相对分子量,用等电聚焦法测定蛋白的等电点.用薄层析方法确定抗凝蛋白与磷脂酰胆碱结合。结果从短尾蝮蛇中分离纯化出的抗凝蛋白是二聚体,相对分子量为24.0×103(非还原)和14.6×103(还原)。等电点为pH 5.2。该蛋白具有精氨酸酯酶活性,能明显地延长活化的部分凝血活酶时间(APTT),其抗凝活性与磷脂结合有关。结论此方法成功地从短尾蝮蛇毒中纯化出一种抗凝蛋白。因其能够与磷脂结合,又具有明显的抗凝活性,因此把该蛋白称为磷脂结合抗凝蛋白(phospholip id-b ind ing anticoagu lation prote in,PBAP)。展开更多
文摘By successive chromatography on CM-Sepharose CL-6B,Sephadex G-100,Heparin-Sepharose CL-6B, a phosphodiesterase (PDE, EC3.1.4.1) was isolated and purified from Guangxi cobra venom. The final preparation was 89 fold pure and had high specific activity. The enzyme showed a single band on SDS-PAGE and IEF. The PDE showed a molecular weight of 110 kD and pI of 7.4. The enzyme consists of 840 amino acid residues and has only a methionine as N-terminal amino acid residues. It was rich in asparagine, leucine and glutamine and poor in sulphur-containing amino acid. It contains 5.38% neutral sugars. It has no acute toxicity and has analgesia effect.
文摘目的从短尾蝮蛇毒中分离纯化一种抗凝蛋白,并对其生化性质进行研究。方法利用阳、阴离子交换、凝胶过滤的方法分离纯化这种抗凝蛋白,用活化部分凝血活酶时间(APTT)测定其抗凝活性,用SDS-PAGE测定其蛋白相对分子量,用等电聚焦法测定蛋白的等电点.用薄层析方法确定抗凝蛋白与磷脂酰胆碱结合。结果从短尾蝮蛇中分离纯化出的抗凝蛋白是二聚体,相对分子量为24.0×103(非还原)和14.6×103(还原)。等电点为pH 5.2。该蛋白具有精氨酸酯酶活性,能明显地延长活化的部分凝血活酶时间(APTT),其抗凝活性与磷脂结合有关。结论此方法成功地从短尾蝮蛇毒中纯化出一种抗凝蛋白。因其能够与磷脂结合,又具有明显的抗凝活性,因此把该蛋白称为磷脂结合抗凝蛋白(phospholip id-b ind ing anticoagu lation prote in,PBAP)。
