您好，欢迎访问山西省农业科学院 机构知识库！
山西省农业科学院机构知识库

全部

  • 全部
  • 标题
  • 学者
  • 机构
  • 关键词

Secretory expression, immunoaffinity purification and metal-binding ability of recombinant metallothionein (ShMT) from freshwater crab Sinopotamon henanense

文献类型： 外文期刊

作者： He, Yongji 1 ; Wang, Lan 2 ; Ma, Wenli 2 ; Lu, Xiaoxia 3 ; Li, Yunlong 1 ; Liu, Jinping 2 ;

作者机构： 1.Shanxi Acad Agr Sci, Inst Agroprod Proc Sci & Technol, 79 Longcheng St, Taiyuan 030031, Shanxi, Peoples R China

2.Shanxi Univ, Sch Life Sci, 92 Wucheng Rd, Taiyuan 030006, Shanxi, Peoples R China

3.Biol Inst Shanxi, 50 Shifan St, Taiyuan 030006, Shanxi, Peoples R China

关键词： Metallothionein; Secretory expression; Immunoaffinity purification; Metal-binding ability; Sinopotamon henanense; Polyol-responsive

期刊名称：ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY （ 影响因子：6.291； 五年影响因子：6.393 ）

ISSN： 0147-6513

年卷期： 2019 年 169 卷

页码：

收录情况： SCI

摘要： Metallothioneins (MTs) are a super-family of ubiquitous, low-molecular-weight, cysteine-rich and metal-binding proteins. They are thought to play a predominant role in mediating metal metabolism and antioxidation. However, the accurate functions of MTs remain unclear in the physiological processes due to native proteins deficiency and little information of their metal-binding character. Freshwater crab Sinopotamon henanense is a decapod crustacean widely distributed in northern China, in which only one MT isoform (ShMT) has been reported so far. In order to shed light on the accurate role of ShMT, a novel recombinant ShMT in native form was over-expressed by phoA secreted expression system in Escherichia colt (E. coli). Then the ShMT proteins were purified using a one-step gentle immunoaffinity chromatography with a polyol-responsive mAb (PR-mAb) to ShMT, which was generated by conventional hybridoma technology followed by ELISA-elution. The Zn-, Cu-, and Cd-ShMT complexes were prepared by recombinant synthesis in metal-enriched media and reconstitution with metal ions, respectively. Further analysis about metal-binding capacity showed recombinant ShMT has high ability to bind Zn, Cu and Cd metals, although the recombinantly expressed and reconstituted metal-ShMT complexes have different metal-to-protein stoichiometry. Moreover, the affinity of recombinant protein for metal ions has been analyzed using competitive reaction with 5, 5-dithiobis (2-nitrobenzoic acid) (DTNB). The results demonstrated the affinity of recombinant ShMT for metals was as follows: Cu > Cd > Zn. In summary, the experimental procedure we have developed facilitates production of recombinant ShMT with native characteristics for further research and the study of metal-binding ability could help further clarify the accurate functions of ShMT.

  • 相关文献
作者其他论文 更多>>
置顶
购物车
反馈

© 京ICP备09089781号-29 主办单位：山西省农业科学院

学术资源： 中国农业科学院 农业专业知识服务系统 农科联盟资源共建共享平台 中国农业科技文献与信息服务平台 中国农业期刊集成服务平台