THE CLONING, EXPRESSION AND PURIFICATION OF RECOMBINANT HUMAN NEURITIN FROM Escherichia coli AND THE PARTIAL ANALYSIS OF ITS NEUROBIOLOGICAL ACTIVITY

被引：4

作者：

Li, Yuanyuan ^{[1
,2
]}

Tang, Juan ^{[3
]}

Zhang, Yunhua ^{[1
,2
]}

Wang, Haiyan ^{[1
,2
]}

Yuan, Wumei ^{[1
,2
]}

Yu, Na ^{[1
,2
]}

Luo, Xing ^{[1
,2
]}

Xu, Xiaoling ^{[4
]}

Huang, Jin ^{[1
,2
]}

Yang, Lei ^{[4
]}

机构：

[1] Shihezi Univ, Sch Med, Key Lab Xinjiang Endem & Ethn Dis, Shihezi, Xinjiang, Peoples R China

[2] Shihezi Univ, Dept Biochem, Shihezi, Xinjiang, Peoples R China

[3] Shihezi Univ, Sch Med, Affiliated Hosp 1, Shihezi, Xinjiang, Peoples R China

[4] Hangzhou Normal Univ, Hangzhou, Zhejiang, Peoples R China

来源：

CELLULAR & MOLECULAR BIOLOGY LETTERS | 2015年 / 20卷 / 05期

关键词：

Neuritin; Expression; Refolding; Purification; PC12; cells; Dorsal root ganglion (DRG); Neurite outgrowth; Escherichia coli; PC12; CELLS; IN-VITRO; CPG15; PLASTICITY; SURVIVAL; NEURONS; GROWTH; NGF;

D O I：

10.1515/cmble-2015-0057

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Neuritin (Nrn1) is a neurotrophic factor that plays various roles in neural development and synaptic plasticity. In this study, the NRN1 gene was cloned and expressed in Escherichia coli and then recombinant neuritin protein was purified so that its neurobiological activity could be evaluated. The protein, which was obtained at a concentration of 0.45 mg/ml and > 90% purity, had the predicted molecular weight of 30 kDa, as determined via sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blot analysis confirmed that an anti-neuritin antibody could recognize the fusion protein. Subsequent functional analyses revealed that recombinant neuritin promoted neurite outgrowth in embryonic chicken dorsal root ganglia and PC12 cells. These results suggest that recombinant neuritin protein could be a valuable tool for inducing neurite regeneration, for instance in cases of spinal cord injury or neurological diseases.

引用

页码：965 / 973

页数：9

共 23 条

[1] Interaction of survival of motor neuron (SMN) and HuD proteins with mRNA cpg15 rescues motor neuron axonal deficits [J].

Akten, Bikem ;

Kye, Min Jeong ;

Hao, Le T. ;

Wertz, Mary H. ;

Singh, Sasha ;

Nie, Duyu ;

Huang, Jia ;

Merianda, Tanuja T. ;

Twiss, Jeffery L. ;

Beattie, Christine E. ;

Steen, Judith A. J. ;

Sahin, Mustafa .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2011, 108 (25) :10337-10342

[2] Differential gene expression in migratory streams of cortical interneurons [J].

Antypa, Mary ;

Faux, Clare ;

Eichele, Gregor ;

Parnavelas, John G. ;

Andrews, William D. .

EUROPEAN JOURNAL OF NEUROSCIENCE, 2011, 34 (10) :1584-1594

[3] Neuritin (cpg15) enhances the differentiating effect of NGF on neuronal PC12 cells [J].

Cappelletti, Graziella ;

Galbiati, Mariarita ;

Ronchi, Cristina ;

Maggioni, Maria Grazia ;

Onesto, Elisa ;

Poletti, Angelo .

JOURNAL OF NEUROSCIENCE RESEARCH, 2007, 85 (12) :2702-2713

[4] Impact of Neuritin 1 (NRN1) Polymorphisms on Fluid Intelligence in Schizophrenia [J].

Chandler, David ;

Dragovic, Milan ;

Cooper, Matthew ;

Badcock, Johanna C. ;

Mullin, Benjamin H. ;

Faulkner, Deb ;

Wilson, Scott G. ;

Hallmayer, Joachim ;

Howell, Sarah ;

Rock, Daniel ;

Palmer, Lyle J. ;

Kalaydjieva, Luba ;

Jablensky, Assen .

AMERICAN JOURNAL OF MEDICAL GENETICS PART B-NEUROPSYCHIATRIC GENETICS, 2010, 153B (02) :428-437

[5]

Chen Xia, 2010, Zhonghua Yan Ke Za Zhi, V46, P978

[6] Production of biologically active forms of recombinant hepcidin, the iron-regulatory hormone [J].

Gagliardo, Bruno ;

Faye, Audrey ;

Jaouen, Maryse ;

Deschemin, Jean-Christophe ;

Canonne-Hergaux, Francois ;

Vaulont, Sophie ;

Sari, Marie-Agnes .

FEBS JOURNAL, 2008, 275 (15) :3793-3803

[7] Characterization of neuritin as a novel angiogenic factor [J].

Han, Dingding ;

Qin, Bo ;

Liu, Guoqing ;

Liu, Tingting ;

Ji, Guoqing ;

Wu, Yanhua ;

Yu, Long .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2011, 415 (04) :608-612

[8] The enhancement of cell adherence and inducement of neurite outgrowth of dorsal root ganglia co-cultured with hyaluronic acid hydrogels modified with Nogo-66 receptor antagonist in vitro [J].

Hou, S ;

Tian, W ;

Xu, Q ;

Cui, F ;

Zhang, J ;

Lu, Q ;

Zhao, C .

NEUROSCIENCE, 2006, 137 (02) :519-529

[9] Expression and purification of the functional ectodomain of human anthrax toxin receptor 2 in Escherichia coli Origami B cells with assistance of bacterial Trigger Factor [J].

Jacquez, Pedro ;

Lei, Ningjing ;

Weigt, David ;

Xiao, Chuan ;

Sun, Jianjun .

PROTEIN EXPRESSION AND PURIFICATION, 2014, 95 :149-155

[10] Neuritin mediates nerve growth factor-induced axonal regeneration and is deficient in experimental diabetic neuropathy [J].

Karamoysoyli, Eugenia ;

Burnand, Rebecca C. ;

Tomlinson, David R. ;

Gardiner, Natalie J. .

DIABETES, 2008, 57 (01) :181-189

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