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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 new separation method of conotoxin based on two-dimensional liquid chromatography (LC-MS), and to understand its constitution characteristics of toxins. Methods The conotoxin toxin was extracted from the wart, and the total protein of conotoxin was extracted. Based on the traditional methods of gel chromatography and reversed-phase chromatography, according to the isoelectric point and hydrophobicity of conotoxin, Proteome Lab TMPF2D) and subjected to two-dimensional separation. Results After the traditional separation method, about 40 bands of conotoxin could be detected, and the separation effect was not obvious. The two-dimensional liquid chromatographic separation method, pI / UV spectrum showed a total of about 200 conotoxin bands were detected. Conclusion Compared with traditional methods, Proteome LabTMPF2D system can separate toxins more rapidly and with higher resolution, which is more conducive to the identification of conotoxin peptides. It also lays a good foundation for the study of structural characteristics and biological activity of downstream sequences.