Soluble expression and one-step purification of a neurotoxin   Huwentoxin-I in Escherichia coli

doi:10.1016/j.pep.2009.02.002

CORC > 北京大学 > 生命科学学院

	Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli
	Che, Nanying ; Wang, Lili ; Gao, Yin ; An, Chengcai
刊名	蛋白质表达与纯化
	2009
关键词	Huwentoxin-1 Neurotoxin Soluble expression Intein N-VSCC Whole-cell patch clamp SPIDER SELENOCOSMIA-HUWENA CALCIUM-CHANNEL BLOCKER OMEGA-CONOTOXIN MVIIA 3-DIMENSIONAL STRUCTURE VENOM PROTEIN INTEIN
DOI	10.1016/j.pep.2009.02.002
英文摘要	Huwentoxin-I (HWTX-I) is a small 33-amino acid neurotoxin from the venom of the Chinese bird spider Ornithoctonus huwena. HWTX-I selectively blocks N-type voltage-sensitive calcium channels (N-VSCCs) and has great potential for clinical application as a novel analgesic without inducing drug tolerance. However, there are still many unsolved issues for this peptide, such as its clinical efficacy in analgesia, anesthesia, and even its potential role in drug rehabilitation. Therefore, large amounts of active recombinant HWTX-I are urgently needed. In this report, we describe a novel and efficient way to produce large amounts of the valuable form in Escherichia coli. HWTX-I was expressed in soluble form as an N-terminal intein fusion product. After affinity purification, a pH shift-induced self-cleavage of the intein released HWTX-I, resulting in a single-column purification of the target protein. The whole-cell patch clamp assay showed that purified HWTX-I has activity similar to another commercialized N-VSCC blocker omega-conotoxin MVIIA. Production of HWTX-I by this method has the major advantages of high efficiency and low cost. (C) 2009 Elsevier Inc. All rights reserved.; Biochemical Research Methods; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; SCI(E); PubMed; 11; ARTICLE; 2; 154-159; 65
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/197681]
专题	生命科学学院
推荐引用方式 GB/T 7714	Che, Nanying,Wang, Lili,Gao, Yin,et al. Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli[J]. 蛋白质表达与纯化,2009.
APA	Che, Nanying,Wang, Lili,Gao, Yin,&An, Chengcai.(2009).Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli.蛋白质表达与纯化.
MLA	Che, Nanying,et al."Soluble expression and one-step purification of a neurotoxin Huwentoxin-I in Escherichia coli".蛋白质表达与纯化 (2009).