中山大学学报自然科学版 ›› 2018, Vol. 57 ›› Issue (2): 137-142.

• 论文 • 上一篇    下一篇

芋螺毒素lt1c的重组表达及镇痛功能

王磊1,曾夏芸2,强媛媛2,任政华2   

  1. 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
  • 收稿日期:2017-08-30 出版日期:2018-03-25 发布日期:2018-03-25
  • 通讯作者: 任政华(1954年生),女; 研究方向: 生物制药;E-mail:renzhh@mail.sysu.edu.cn

Recombinant expression and functional analysis of conotoxin lt1c

WANG Lei1, ZENG Xiayun2, QIANG Yuanyuan2,REN Zhenghua2   

  1. 1.Department of Pharmaceutical Engineering, College of Materials and Energy, South China Agricultural University, Guangzhou 510642, China; 
    2. National Engineering Research Center of South China Sea Marine Biotechnology, Guangzhou 510642, China
  • Received:2017-08-30 Online:2018-03-25 Published:2018-03-25

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6 μmol/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。

关键词: A-超家族毒素, 芋螺毒素, 重组表达, 镇痛

Abstract:

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 6 μmol/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.

Key words: A-superfamily, conotoxin, recombinant expression, analgesic

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