Lin Shinne-Ren, Chang Long-Sen, Chang Kee-Lung
School of Chemistry, Kaohsiung Medical University, Taiwan, ROC.
J Protein Chem. 2002 Feb;21(2):81-6. doi: 10.1023/a:1014520126856.
Six cardiotoxins (CTXs) and one cardiotoxin-like basic protein (CLBP) from Naja naja atra (Taiwan cobra) venom were separated by a SP-Sephadex C-25 column. CTXn and CTXI were well separated by eluting with ammonium acetate buffer, and the separation of CLBP from CTXIV and CTXV mixtures was achieved using sodium phosphate buffer. These findings suggest a differential interaction of CTXs with the chromatographic matrix using different buffer systems. Chemical modification studies on cationic residues of CTXI suggested that there was no single lysine or arginine residue exclusively responsible for its biological activity. Moreover, it was found that the cytotoxicity and hemolytic sites of CTXI could be dissociated by chemical modifications. It suggests the potentiality for preparing toxin derivatives in which a specific activity is retained.
用SP - Sephadex C - 25柱从眼镜蛇(台湾眼镜蛇)毒液中分离出六种心脏毒素(CTXs)和一种类心脏毒素碱性蛋白(CLBP)。通过用醋酸铵缓冲液洗脱,CTXn和CTXI得到了很好的分离,并且使用磷酸钠缓冲液实现了从CTXIV和CTXV混合物中分离出CLBP。这些发现表明,使用不同的缓冲系统,CTXs与色谱基质存在差异相互作用。对CTXI阳离子残基的化学修饰研究表明,没有单个赖氨酸或精氨酸残基专门负责其生物活性。此外,发现CTXI的细胞毒性和溶血位点可通过化学修饰解离。这表明制备保留特定活性的毒素衍生物具有潜力。