化学合成虎纹捕鸟蛛毒素-Ⅰ的色谱行为及分离纯化  

Chromatographic Behavior and Purification of Synthetic Huwentoxin-Ⅰ

作　　者：王贤纯[1] 梁宋平[1] 

机构地区：[1]湖南师范大学生命科学学院,湖南长沙410081

出　　处：《色谱》2001年第2期101-104,共4页Chinese Journal of Chromatography

基　　金：国家 8 63高技术发展计划资助! (10 3 13 0 1 0 6)

摘　　要：利用反相高效液相色谱 (RP HPLC)和离子交换色谱 (IEC)比较研究了固相化学合成的多肽类神经毒素虎纹捕鸟蛛毒素 Ⅰ (HWTX Ⅰ )与其对应的天然产物的色谱行为差异及分离纯化方法。发现在完全相同的实验条件下 ,复性处理后的合成HWTX Ⅰ的谱峰有别于还原变性后又复性的天然HWTX Ⅰ的谱峰 ,主要表现为扩散度较大 ,对称性较差 ,说明合成产物存在更大分子构象的不均一性。推测除了在合成HWTX Ⅰ复性过程中有少数分子发生二硫键错配外 ,还有部分分子在固相合成中发生了消旋作用 。A comparative study was made to investigate the chromatographic behavior and purification of chemically synthesized and native polypeptide neurotoxin huwentoxin Ⅰ(HWTX Ⅰ) by means of reversed phase HPLC and ion exchange chromatography The results showed that the synthetic HWTX Ⅰ in crude product has a longer retention time than the native HWTX Ⅰ,but has the same retention time as the denatured HWTX Ⅰ, suggesting it being in a state of complete denaturation which was confirmed by the MALDI TOF MS analysis It has also been discovered that, before complete purification, the synthetic HWTX Ⅰ, after oxidative treatment, showed wider and more asymmetric peaks than the native HWTX Ⅰ that was denatured and then renatured under the same experimental conditions, which suggests that there is even greater conformational unhomogeneity in the synthetic HWTX Ⅰ It was inferred that, in addition to the wrong disulfide bonds formation during oxidative folding, the racemization during chemical synthesis resulted in more stereoisomers of synthetic HWTX Ⅰ It was needed for the renatured synthetic peptides to be purified using different methods alternatively

关 键 词：色谱 纯化 固相合成 分离 虎纹捕岛蛛毒素-Ⅰ 神经毒素 

分 类 号：R996.3[医药卫生—毒理学] O658.1[医药卫生—药学]