多棘蜈蚣钙通道毒素多肽CaTX-Ssmd1原核表达及功能鉴定  

Prokaryotic expression and functional identification of Scolopendra subspinipes multidens calcium channel toxin polypeptide CaTX-Ssmd1

作　　者：尹世金[1] 肖敏 向琪琳 李昱烨 刘柯驿 赵倩茹 YIN Shijin;XIAO Min;XIANG Qilin;LI Yuye;LIU Keyi;ZHAO Qianru(School of Pharmaceutical Sciences,South-Central Minzu University,Wuhan 430074,China)

机构地区：[1]中南民族大学药学院,武汉430074

出　　处：《中南民族大学学报（自然科学版）》2023年第3期313-318,共6页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：国家自然科学基金资助项目(81373379,32000685);省级大学生创新训练项目(SCX2242)。

摘　　要：为了检测广西多棘蜈蚣毒腺转录组文库新基因CaTX-Ssmd1编码的多肽是否具有阻断钙离子通道蛋白的生物活性,将构建的pGEX-4T-1-CaTX-Ssmd1原核表达载体转化Escherichia coli Rosetta(DE3),诱导表达产物经酶切和RP-HPLC分离后得到CaTX-Ssmd1成熟多肽.质谱检测显示CaTX-Ssmd1多肽分子量与理论值一致;钙成像实验表明CaTX-Ssmd1多肽能有效减弱高钾外液灌流激发的小鼠背根神经节(dorsal root ganglion,DRG)细胞[Ca^(2+)]i信号.结论:初步验证了原核表达纯化的CaTX-Ssmd1多肽为新发现的电压门控性钙通道(voltage gated calcium channels,VGCC)阻断剂,丰富了多棘蜈蚣生物活性多肽资源库.To test whether the polypeptide encoded by the new gene CaTX-Ssmd1 in the venom gland transcriptome library of Scolopendra subspinipes multidens has the activity to block calcium channel protein,the prokaryotic expression vector pGEX-4T-1-CaTX-Ssmd1 was transformed into Escherichia coli Rosetta(DE3).The mature CaTX-Ssmd1 polypeptide was obtained from the induced expression product after digestion and RP-HPLC separation.Mass spectrometry analysis showed that the molecular weight of CaTX-Ssmd1 polypeptide was consistent with the theoretical value.Calcium imaging experiment shows that CaTX-Ssmd1 polypeptide can effectively attenuate[Ca^(2+)]i signal of mouse dorsal root ganglion(DRG)cells stimulated by high potassium external liquid perfusion.Conclusions:This study preliminarily verifies that the prokaryotic expression and purification of CaTX-Ssmd1 polypeptide is a newly discovered voltage gated calcium channels(VGCC)blocker,which enriches the resource pool of bioactive polypeptides of Scolopendra subspinipes multidens.

关 键 词：多棘蜈蚣 多肽CaTX-Ssmd1 原核表达 钙通道阻断剂 

分 类 号：R931.74[医药卫生—生药学]