福建产舟山眼镜蛇毒细胞毒素的快速分离纯化及鉴定  

Purification and Identification of Cytotoxin Isoforms from the Venom of Naja atra

作　　者：张明芳[1] 许云禄[2] 刘广芬[2] 

机构地区：[1]福建医科大学生理学与病理生理学系,福州350004 [2]福建医科大学蛇毒研究所,福州350004

出　　处：《福建医科大学学报》2004年第1期1-4,共4页Journal of Fujian Medical University

基　　金：福建省自然科学基金资助项目 ( C0 3 10 0 2 6)

摘　　要：目的　从舟山眼镜蛇毒中分离纯化细胞毒素 (CTX) ,并鉴定其理化性质。　方法　采用 SP- SephadexC- 2 5阳离子交换色谱及 Sephasil Peptide C1 8反相高效液相色谱法从舟山眼镜蛇毒中分离纯化 CTX,SDS- PAGE(Tris- Tricine系统 )鉴定纯度 ,Edman降解法测定 N端氨基酸序列。　结果　粗毒经阳离子交换色谱 ,得到 1 4个蛋白峰 ,其中第 ～ 峰具 CTX活性 ;再分别经反相色谱纯化 ,得到 4个 CTX,总得率为 32 .4 8% ;SDS- PAGE显示为均一蛋白 ,分子量依次为 :7.2 8,7.33,7.2 4和 7.38k D;测定它们 N端 2 0个氨基酸序列。　结论　采用阳离子交换和反相高效液相色谱可快速、高效地从眼镜蛇毒中获得 4个Objective To isolate and purify cytotoxin isoforms from Naja atra and further appraise its properties. Methods Cytotoxins were purified from Naja atra venom by successive separation on a SP|Sephadex C|25 column and a Sephasil Peptide C-{18} column. The purity of cytotoxins was appraised by SDS|polyacrylamide gel electrophoresis(Tris|Tricine system). N|terminal amino acid sequences of cytotoxins were determined by Edman degradation. Results Ion|exchange chromatography of lyophilized crude venom yielded 14 fractions. By reverse phase high performance liquid chromatography, four cytotoxins were purified from fractions Ⅹ~ⅩⅢ which have cytotoxic activities. The total yield of cytotoxin was 32 48%. SDS|PAGE and N|terminal sequencing analysis of them showed that molecular were 7 28, 7 33, 7 24 and 7 38 kD orderly. Conclusion Four cytotoxin isoforms were obtained from Naja atra venom. It is proved that the method of a successive cation|exchange and RP|HPLC is efficient in cytotoxin purification.

关 键 词：眼镜蛇毒液类 细胞毒素类 色谱法 高压液相 色谱法 离子交换 氨基酸序列 阳离子 

分 类 号：R99[医药卫生—毒理学]