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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Soluble expression, purification and the role of C-terminal glycine residues in scorpion toxin BmK AGP-SYPU2

  • Zhang, Rong (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Cui, Yong (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Zhang, Xi (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Yang, Zhuo (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Zhao, Yongshan (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Song, Yong-Bo (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Wu, Chunfu (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University) ;
  • Zhang, Jinghai (School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University)
  • Received : 2010.10.07
  • Accepted : 2010.11.05
  • Published : 2010.12.31
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Abstract

The existence of glycine residues in long-chain scorpion toxins has been well documented. However, their role as analgesics has not been evaluated. To address this issue, we investigated the functional role of glycines in the C-terminal end of Chinese-scorpion toxin from Buthus martensii Karsch (BmK AGP-SYPU2) using site-directed mutagenesis and analgesic activity assays. Recombinant BmK AGP-SYPU2 and its mutants were efficiently expressed in E. coli and purified to homogeneity using immobilized metal ion affinity chromatography (IMAC) and cation exchange chromatography. The mouse-twisting test was used to detect the analgesic activity of BmK AGP-SYPU2 and its mutants. As a result, we identified glycines at the C-terminal end that, when altered, significantly affected analgesic activity. Also, Mut6566 was significantly decreased compared to BmK AGP-SYPU2. These data indicate that the glycines at the C-terminal end are important for the analgesic activity of BmK AGP-SYPU2.

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References

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