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目的从广东眼镜蛇粗毒中分离纯化出色谱纯的低分子量多肽并对其部分性质进行鉴定。方法采用化学沉淀法对蛇粗毒进行预处理,经Sephadex G-50凝胶过滤和UND Sphere S阳离子交换层析得到高纯度低分子量多肽,用SDS-PAGE及HPLC对产物纯度进行鉴定,并用SDS-PAGE和质谱法测定分子量,采用蛙心灌流法考察产物是否具有心脏毒性。结果结果表明,经分离纯化后得到两种低分子量多肽GI2和GI3,均为单一组分,可达到色谱纯,SDS-PAGE法测得GI2分子量为14 300 Da,质谱法测得GI3分子量为6 725.8 Da,仅GI2具有心脏毒性,收率分别为9.5%和10.1%。结论通过化学沉淀预处理,并经2次柱色谱可从广东眼镜蛇毒中分离得到两种色谱纯的低分子量多肽,且收率较高。
OBJECTIVE To isolate and purify the chromatographic pure low molecular weight peptides from the cobra venom of Guangdong Cobra and identify some of their properties. Methods Snake venom was pretreated by chemical precipitation method. High purity and low molecular weight peptides were obtained by Sephadex G-50 gel filtration and UND Sphere S cation exchange chromatography. The purity of the product was identified by SDS-PAGE and HPLC. -PAGE and mass spectrometry determination of molecular weight, using frog heart perfusion method to examine whether the product has cardiotoxicity. The results showed that two kinds of low molecular weight peptides GI2 and GI3 were obtained after isolation and purification, which were single components and reached chromatographic purity. The molecular weight of GI2 was 14 300 Da determined by SDS-PAGE and the molecular weight of GI3 was 6 725.8 Da, only GI2 with cardiotoxicity, the yield was 9.5% and 10.1%. Conclusions Two kinds of chromatographic pure low molecular weight peptides were isolated from Guangdong cobra venom by chemical precipitation pretreatment and twice column chromatography with high yield.