Expression in Escherichia coli of fusion protein comprising α-conotoxin TxIB and preservation of selectivity to nicotinic acetylcholine receptors in the purified product

2017 ◽  
Vol 91 (2) ◽  
pp. 349-358 ◽  
Author(s):  
Jinpeng Yu ◽  
Xiaopeng Zhu ◽  
Yang Yang ◽  
Sulan Luo ◽  
Dongting Zhangsun
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
Nicotinic Acetylcholine ◽  
Expression In Escherichia Coli

scholarly journals α-Conotoxin as Potential to α7-nAChR Recombinant Expressed in Escherichia coli

Marine Drugs ◽  
2020 ◽  
Vol 18 (8) ◽  
pp. 422
Author(s):  
Yanli Liu ◽  
Yifeng Yin ◽  
Yunyang Song ◽  
Kang Wang ◽  
Fanghui Wu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
High Performance ◽  
Cognitive Disorders ◽  
Coli Strain ◽  
Xenopus Laevis Oocytes ◽  
Obvious Effect ◽  
Nicotinic Acetylcholine ◽  
Α7 Nachr

α7 nicotinic acetylcholine receptors (nAChR) is an important nicotinic acetylcholine receptors subtype and closely associated with cognitive disorders, such as Alzheimer’s and schizophrenia disease. The mutant ArIB (V11L, V16A) of α-conotoxin ArIB with 17-amino acid residues specifically targets α7 nAChR with no obvious effect on other nAChR subtypes. In the study, the synthetic gene encoding mature peptide of ArIB and mutant ArIB (V11L, V16A) carried a fusion protein Trx and 6 × His-tag was separately inserted in pET-32a (+) vector and transformed into Escherichia coli strain BL21(DE3) pLysS for expression. The expressions of Trx-ArIB-His6 and Trx-ArIB (V11L, V16A)-His6 were soluble in Escherichia coli, which were purified by Ni-NTA affinity chromatography column and cleaved by enterokinase to release rArIB and rArIB (V11L, V16A). Then, rArIB and rArIB (V11L, V16A) were purified by high-performance liquid chromatography (HPLC) and identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). Bioactivity of rArIB and rArIB (V11L, V16A) was assessed by two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing human nAChR subtypes. The results indicated that the yield of the fusion proteins was approximately 50 mg/L and rArIB (V11L, V16A) antagonized the α7 nAChR subtype selectively with 8-nM IC50. In summary, this study provides an efficient method to biosynthesize α-conotoxin ArIB and rArIB (V11L, V16A) in Escherichia coli, which could be economical to obtain massively bioactive disulfide-rich polypeptides at fast speed.

Antisera against an acetylcholine receptor α3 fusion protein bind to ganglionic but not to brain nicotinic acetylcholine receptors

FEBS Letters ◽  
1989 ◽  
Vol 257 (2) ◽  
pp. 393-399 ◽  
Author(s):  
Ralf Schoepfer ◽  
Stanley W. Halvorsen ◽  
William G. Conroy ◽  
Paul Whiting ◽  
Jon Lindstrom
Keyword(s):  
Fusion Protein ◽  
Acetylcholine Receptor ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
Nicotinic Acetylcholine

Nicotinic acetylcholine receptors in the patients with Parkinson's disease: 5IA-SPECT

2005 ◽  
Vol 25 (1_suppl) ◽  
pp. S586-S586 ◽  
Author(s):  
Kazuo Hashikawa ◽  
Hidefumi Yoshida ◽  
Nobukatsu Sawamoto ◽  
Shigetoshi Takaya ◽  
Chihiro Namiki ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Nicotinic Acetylcholine Receptors ◽  
Acetylcholine Receptors ◽  
Nicotinic Acetylcholine
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