Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli

α-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...
Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Xiaopeng Zhu [verfasserIn] Jianpeng Bi [verfasserIn] Jinpeng Yu [verfasserIn] Xiaodan Li [verfasserIn] Yaning Zhang [verfasserIn] Dongting Zhangsun [verfasserIn] Sulan Luo [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2016

Schlagwörter:	α-conotoxin LvIA recombinant expression fusion protein nAChRs electrophysiology pain assay

Übergeordnetes Werk:	In: Marine Drugs - MDPI AG, 2005, 14(2016), 1, p 11
Übergeordnetes Werk:	volume:14 ; year:2016 ; number:1, p 11

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3390/md14010011

Katalog-ID:	DOAJ085283789

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authorswithroles_txt_mv	Xiaopeng Zhu @@aut@@ Jianpeng Bi @@aut@@ Jinpeng Yu @@aut@@ Xiaodan Li @@aut@@ Yaning Zhang @@aut@@ Dongting Zhangsun @@aut@@ Sulan Luo @@aut@@
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callnumber-first	Q - Science
author	Xiaopeng Zhu
spellingShingle	Xiaopeng Zhu misc QH301-705.5 misc α-conotoxin LvIA misc recombinant expression misc fusion protein misc nAChRs misc electrophysiology misc pain assay misc Biology (General) Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
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topic_title	QH301-705.5 Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli α-conotoxin LvIA recombinant expression fusion protein nAChRs electrophysiology pain assay
topic	misc QH301-705.5 misc α-conotoxin LvIA misc recombinant expression misc fusion protein misc nAChRs misc electrophysiology misc pain assay misc Biology (General)
topic_unstemmed	misc QH301-705.5 misc α-conotoxin LvIA misc recombinant expression misc fusion protein misc nAChRs misc electrophysiology misc pain assay misc Biology (General)
topic_browse	misc QH301-705.5 misc α-conotoxin LvIA misc recombinant expression misc fusion protein misc nAChRs misc electrophysiology misc pain assay misc Biology (General)
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title	Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
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title_full	Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
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title_sort	recombinant expression and characterization of α-conotoxin lvia in escherichia coli
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title_auth	Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
abstract	α-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.
abstractGer	α-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.
abstract_unstemmed	α-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.
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title_short	Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli
url	https://doi.org/10.3390/md14010011 https://doaj.org/article/a4beb6ebc526440b9e7c5e7a6f83f1d0 http://www.mdpi.com/1660-3397/14/1/11 https://doaj.org/toc/1660-3397
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Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli

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