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KMID : 0624620100430120801
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BMB Reports
2010 Volume.43 No. 12 p.801 ~ p.806
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Soluble expression, purification and the role of C-terminal glycine residues in scorpion toxin BmK AGP-SYPU2
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Zhang Rong
Cui Yong
Zhang Xi
Yang Zhuo
Zhao Yongshan
Song Yongbo
Wu Chunfu
Zhang Jinghai
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Abstract
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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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KEYWORD
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Analgesic activity, BmK AGP-SYPU2, C-terminal glycine residues, Purification, Soluble expression
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