芋螺毒素lt1c的重组表达及镇痛功能  

Recombinant expression and functional analysis of conotoxin lt1c

作　　者：王磊 曾夏芸 强媛媛 任政华 WANG Lei;ZENG Xiayun;ZENG Xiayun;REN Zhenghua(Department of Pharmaceutical Engineering,College of Materials and Energy,South China Agricultural University,Guangzhou 510642,China;National Engineering Research Center of South China Sea Marine Biotechnology,Guangzhou 510642,China)

机构地区：[1]华南农业大学材料与能源学院制药工程系,广东广州510642 [2]南海海洋生物技术国家工程研究中心,广东广州510642

出　　处：《中山大学学报（自然科学版）》2018年第2期137-142,共6页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：广东省自然科学基金(2015A030313191)

摘　　要：lt1c是从信号芋螺毒管中发现的一种A超家族毒素多肽,为深入研究其功能,采用pTRX表达系统将lt1c与硫氧还蛋白基因进行融合,转入大肠杆菌进行表达后,通过超声破碎、亲和层析、酶切和凝胶过滤层析等方法,最终得到目的蛋白。组织水平的青蛙坐骨神经-腓肠肌活性研究表明,重组表达的lt1c在6μmol/L浓度下,30 min内完全抑制肌肉的收缩,且其作用可逆。进一步的功能研究发现,lt1c能够抑制热板疼痛模型小鼠的舔足反应,提示A超家族芋螺毒素lt1c可能具有一定的镇痛活性。该研究为筛选新的多肽镇痛药物奠定了基础。Conotoxin lt1c was identified from the cDNA library of venom gland of Conus litteratus.To obtain enough lt1c and investigate the bioactivities,conotoxin lt1c was recombinant and expressed in Escherichia coli by fusion with TRX.Conotoxin lt1c was purified by affinity chromatography,enzyme digestion and gel filtration chromatography.The recombinant lt1c showed obvious inhibition activity on the neuromuscular transmission on the frog sciatic/cutaneus pectoris preparation at the concentration of 6μmol/L in 30 min.lt1c appeared potent and longer-lasting analgesics in the hotplate pain model mice.Additionally,lt1c showed toxicity to mice by intraventricular injection.The successful expression of lt1c with potential analgesicactivity provided the possibility for further pharmaceutical research.

关 键 词：A-超家族毒素 芋螺毒素 重组表达 镇痛 

分 类 号：Q78[生物学—分子生物学] R965[医药卫生—药理学]