摘要
为了探讨质谱分析在合成多肽氧化复性和分离纯化研究中的应用 ,用固相多肽合成方法合成ω -芋螺毒素MⅦA ,在含谷胱甘肽的缓冲体系中进行氧化复性后 ,经离子交换和RP HPLC分离纯化。利用基质辅助激光解吸电离飞行时间质谱 (MALDI TOF MS)和电喷雾串联质谱 (ESI MS/MS)分析ω -芋螺毒素MⅦA氧化复性和分离纯化的效果 ,最后用电生理学实验测定复性ω -芋螺毒素MⅦA的生理活性。其结果表明 ,获得的ω -芋螺毒素MⅦA纯化复性样品具有与天然ω
In order to investigate the application of mass spectrometry (MS) in the renaturation and purification of synthetic peptides,ω conotoxin MⅦA was chemically synthesized by a solid phase method.The sample was oxidatively treated in a glutathione containing buffer,and then purified by ion exchange chromatography and RP HPLC.The renaturation and purification efficiencies of synthesized ω conotoxin MⅦA were subsequently analyzed by MALDI TOF MS and tandem electrospray mass spectrometry (ESI MS/MS).Meanwhile,using an electrophysiological method,its biological activity was also measured.Mass spectrometric and electrophysiological analysis revealed that renatured and purified ω conotoxin MⅦA possesses the same conformation and physiological activity as those of natural ω conotoxin MⅦA.
基金
湖南省自然科学基金资助项目 ( 0 1JJY2 0 2 8)
关键词
ω-芋螺毒素MⅦA
氧化复性
质谱分析
conotoxin MⅦA
Oxidative renaturation
Mass spectrometric analysis
