疣缟芋螺毒素的二维液相色谱分离方法  

Conotoxin extraction and fractionation by the two-dimensional liquid chromatography from Conus lividus venom

作　　者：吴勇[1] 朱晓鹏[1] 胡远艳[1] 长孙东亭[1] 罗素兰[1] 

机构地区：[1]热带生物资源教育部重点实验室海南大学生物技术实验中心海南大学海洋学院,海南海口570228

出　　处：《中国海洋药物》2010年第1期36-40,共5页Chinese Journal of Marine Drugs

基　　金：国家863计划(2007AA02Z114);国家自然科学基金(30860368);重大新药创制国家科技重大专项课题(2009ZX09103-644);海南大学创新团队(hd09xm02);重点科研项目(hd09xm15);科研基金项目(kyjj0617)

摘　　要：目的建立一种新的基于二维液相色谱技术的疣缟芋螺毒素分离方法,了解其毒素组构成特点。方法从疣缟芋螺毒管中提取芋螺毒素总肽,在传统的凝胶色谱和反相色谱方法的基础上,根据芋螺毒素的等电点和疏水性,利用目标蛋白快速分离系统(Proteome Lab TMPF2D),对其进行二维分离。结果经过传统分离方法,能够检测得到40个左右的芋螺毒素条带,且分离效果不明显。而通过二维液相色谱分离方法,pI/UV图谱显示共检测到约200个芋螺毒素条带。结论同传统分离方法相比,采用Proteome LabTMPF2D系统对毒素分离更快速、分辨率更高,因此更利于鉴定芋螺毒素肽组分,也为下游序列结构特征与生物活性研究奠定了良好的基础。Objective To establish a separation protocol based on the two-dimensional liquid chromatography and to identify the conotoxin components from Conus lividus crude venoms.Methods The total crude venom was extracted from C.lividus venom ducts,which was fractionated by a novel two-dimensional liquid chromatography （ProteomeLabTMPF2D）after the initial gel filtration chromatography separation.Results There were only 40 conopeptide peaks detected when the traditional HPLC separation was applied for C.lividus crude venom fractionation,while about 200 conopeptides were detected when the two-dimensional liquid chromatography was applied.Conclusion Compared with the traditional separational protocol,crude venom fractionation of Conus lividus with the ProteomeLabTM PF2D system showed better resolution results with high efficiency and high-sensitivity.Thereby the results in this paper would facilitate the downstream identification of conopeptides＇ sequence and bioactivity.

关 键 词：疣缟芋螺 二维液相色谱分离 芋螺毒素 

分 类 号：R917[医药卫生—药物分析学] R991[医药卫生—药学]