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首页> 外文期刊>Journal of natural toxins >Purification, cDNA cloning and molecular characteristic of a fibrinolytic enzyme from the venom of Agkistrodon acutus.
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Purification, cDNA cloning and molecular characteristic of a fibrinolytic enzyme from the venom of Agkistrodon acutus.

机译：cut蛇毒中纤溶酶的纯化，cDNA克隆和分子特征。

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摘要

A nonglycoprotein-like fibrinolytic enzyme ((FIB-I) was purified from the crude venom of Agkistrodon acutus by CM-Sepharose CL-6B and DEAE-Sepharose CL-6B ion exchange chromatography and then by FPLC through Superose 12 gel filtration. Its molecular weight is about 23 kDa and isoelectric point is near 6.0. It not only has fibrinolytic and caseinolytic activity, but also can hydrolyze BAEE. The local hemorrhagic activity was found in mice after the subcutaneous injection of this enzyme. EDTA can inhibit its fibrinolytic activity completely, but PMSF and arrowhead proteinase inhibitor have no such obvious inhibitory effect, thus implying that FIB-I is a metalloproteinase. The N-terminal ten amino acid residues 'STEFQRYMEI' of FIB-I was elucidated. A full-length cDNA gene of this enzyme was cloned by using RT-PCR from the total RNA extracted from the snake venom gland and FIB-I was expressed in E. coli. Having analyzed the sequence, we found that it had a typical zinc-chelating characteristic as 'HEXXHXXGXXHD.'
机译：通过CM-Sepharose CL-6B和DEAE-Sepharose CL-6B离子交换色谱，然后通过FPLC通过Superose 12凝胶过滤从a蛇粗蛇毒的粗毒中纯化出非糖蛋白样纤溶酶（（FIB-1））。重量约23 kDa，等电点接近6.0，不仅具有纤溶和酪蛋白溶解活性，而且还可以水解BAEE，皮下注射该酶后在小鼠体内具有局部止血活性，EDTA可以完全抑制其纤溶活性。 ，但PMSF和箭头蛋白酶抑制剂没有明显的抑制作用，因此暗示FIB-1是一种金属蛋白酶，阐明了FIB-1的N-末端十个氨基酸残基“ STEFQRYMEI”，该cDNA的全长cDNA基因利用RT-PCR技术从蛇毒腺中提取总RNA，并在大肠杆菌中表达FIB-I，分析其序列，发现其具有典型的锌螯合特性。标记为“ HEXXHXXGXXHD”。

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