Site-directed mutagenesis of the toxin from the Chinese scorpion Buthus martensii Karsch (BmKAS): Insight into sites related to analgesic activity

被引:0
作者
Yong Cui
Yong-Bo Song
Lin Ma
Yan-Feng Liu
Guo-Dong Li
Chun-Fu Wu
Jing-Hai Zhang
机构
[1] Shenyang Pharmaceutical University,School of Life Science and Bio
[2] Shenyang Pharmaceutical University,Pharmaceutics
来源
Archives of Pharmacal Research | 2010年 / 33卷
关键词
Analgesic activity; BmKAS; Homology modeling; Site directed mutagenesis; Structure-function relationship;
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摘要
This study utilized the E. coli expression system to investigate the role of amino acid residues in toxin from the Chinese scorpion — Buthus martensii Karsch (BmKAS). To evaluate the extent to which residues of the toxin core contribute to its analgesic activity, ten mutants of BmKAS were obtained by PCR. Using site-directed mutagenesis, all of these residues were substituted with different amino acids. This study represents a thorough mapping and elucidation of the epitopes that form the molecular basis of the toxin’s analgesic activity. Our results showed large mutant-dependent differences that emphasize the important roles of the studied residues.
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页码:1633 / 1639
页数:6
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[1]  
Bessette P. H.(1999)Efficient folding of proteins with multiple disulfide bonds in the Escherichia coli cytoplasm Proc. Natl. Acad. Sci. U.S.A. 96 13703-13708
[2]  
Aslund F.(2002)Structure of variant 2 scorpion toxin from Centruroides sculpturatus Ewing Protein Sci. 11 479-486
[3]  
Beckwith J.(2004)Structural mechanism governing cis and trans isomeric states and an intramolecular switch for cis/trans isomerization of a non-proline peptide bond observed in crystal structures of scorpion toxins J. Mol. Biol. 341 1189-1204
[4]  
Georgiou G.(1989)Site-directed mutagenesis by overlap extension using the polymerase chain reaction Gene 77 51-59
[5]  
Cook W. J.(2004)Molecular basis of the high insecticidal potency of scorpion alpha-toxins J. Biol. Chem. 279 31679-31686
[6]  
Zell A.(2007)The unique pharmacology of the scorpion alpha-like toxin Lqh3 is associated with its fexible C-tail FEBS J. 274 1918-1931
[7]  
Watt D. D.(1989)Structure/activity relationships of scorpion alpha-toxins. Multiple residues contribute to the interaction with receptors Eur. J. Biochem. 181 381-390
[8]  
Ealick S. E.(1999)NMR structures and activity of a novel alpha-like toxin from the scorpion Leiurus quinquestriatus hebraeus J. Mol. Biol. 285 1749-1763
[9]  
Guan R. J.(1993)PROCHECK: a program to check the stereochemical quality of protein structures J. Appl. Crystallogr. 26 283-291
[10]  
Xiang Y.(2005)Expression of the standard scorpion alpha-toxin AaH II and AaH II mutants leading to the identification of some key bioactive elements Biochim. Biophys. Acta 1723 91-99