Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
Abstractα-Conotoxin LvIA is derived from Conus lividus, native to Hainan, and is the most selective inhibitor of α3β2 nicotinic acetylcholine receptors (nAChRs) known to date. In this study, an efficient approach for the production of recombinant α-Conotoxin LvIA is described. Tandem repeats of a LvIA gene fragment were constructed and fused with a KSI gene and a His6 tag in a Escherichia coli (E. coli) expression vector pET-31b(+). The recombinant plasmids were transformed into E. coli and were found to express well. The KSI-(LvIA)n-His6 fusion protein was purified by metal affinity chromatography and then cleaved with CNBr to release recombinant LvIA (rLvIA). High yields of fusion protein ranging from 100 to 500 mg/L culture were obtained. The pharmacological profile of rLvIA was determined by two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing rat nAChR subtypes. The rLvIA antagonized the α3β2 nAChR subtype selectively with a nano-molar IC50. The rLvIA was analgesic in a mouse hot-plate test model of pain. Overall, this study provides an effective method to synthesize α-conotoxin LvIA in an E. coli recombinant expression system, and this approach could be useful to obtain active conopeptides in large quantity and at low cost. View Full-Text
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Zhu, X.; Bi, J.; Yu, J.; Li, X.; Zhang, Y.; Zhangsun, D.; Luo, S. Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli. Mar. Drugs 2016, 14, 11.
Zhu X, Bi J, Yu J, Li X, Zhang Y, Zhangsun D, Luo S. Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli. Marine Drugs. 2016; 14(1):11.Chicago/Turabian Style
Zhu, Xiaopeng; Bi, Jianpeng; Yu, Jinpeng; Li, Xiaodan; Zhang, Yaning; Zhangsun, Dongting; Luo, Sulan. 2016. "Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli." Mar. Drugs 14, no. 1: 11.
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