Jump to ContentJump to Main Navigation
Show Summary Details
More options …

Cellular and Molecular Biology Letters

Online
ISSN
1689-1392
See all formats and pricing
More options …
Volume 20, Issue 5

Issues

The cloning, expression and purification of recombinant human neuritin from Escherichia coli and the partial analysis of its neurobiological activity

Yuanyuan Li
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Juan Tang
  • The First Affiliated Hospital of Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Yunhua Zhang
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Haiyan Wang
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Wumei Yuan
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Na Yu
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Xing Luo
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Xiaoling Xu / Jin Huang
  • Corresponding author
  • The Key Laboratory of Xinjiang Endemic & Ethnic Diseases and Department of Biochemistry, Shihezi University School of Medicine, Shihezi, Xinjiang, P. R. China
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Lei Yang
Published Online: 2016-03-05 | DOI: https://doi.org/10.1515/cmble-2015-0057

Abstract

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.

Keywords: Neuritin; Expression; Refolding; Purification; PC12 cells; Dorsal root ganglion (DRG); Neurite outgrowth; Escherichia coli

References

  • 1. Chandler, D., Dragovic, M., Cooper, M., Badcock, J.C., Mullin, B.H., Faulkner, D. and Wilson, S.G. Impact of neuritin 1 (NRN1) polymorphisms on fluid intelligence in schizophrenia. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 153B (2010) 428-437.Google Scholar

  • 2. Putz, U., Harwell, C. and Nedivi, E. Soluble CPG15 expressed during early development rescues cortical progenitors from apoptosis. Nat. Neurosci. 8 (2005) 322-331.CrossrefGoogle Scholar

  • 3. Min, S., Jian-Bo, L., Hong-Man, Z., Ling-Fei, Y., Min, X. and Jia-Wei, C. Neuritin is expressed in Schwann cells and down-regulated in apoptotic Schwann cells under hyperglycemia. Nutr. Neurosci. 15 (2012) 264-270.CrossrefWeb of ScienceGoogle Scholar

  • 4. Antypa, M., Faux, C., Eichele, G., Parnavelas, J.G. and Andrews, W.D. Differential gene expression in migratory streams of cortical interneurons. Eur. J. Neurosci. 34 (2011) 1584-1594.CrossrefWeb of ScienceGoogle Scholar

  • 5. Zito, A., Cartelli, D., Cappelletti, G., Cariboni, A., Andrews, W., Parnavelas, J., Poletti, A. and Galbiati, M. Neuritin 1 promotes neuronal migration. Brain Struct. Funct. 219 (2014) 105-118.Web of ScienceGoogle Scholar

  • 6. Chen, X., Chen, J., Tian, C., Lin, J. and Wang, Y. Study of CPG15 expression in the visual cortex of normal and monocular form deprived development rats. Zhonghua Yan Ke Za Zhi 46 (2010) 978-983.Google Scholar

  • 7. Lee, W-C.A. and Nedivi, E. Extended plasticity of visual cortex in darkreared animals may result from prolonged expression of cpg15-like genes. J. Neurosci. 22 (2002) 1807-1815.Google Scholar

  • 8. Akten, B., Kye, M.J., Hao, L.T., Wertz, M.H., Singh, S., Nie, D., Huang, J., Merianda, T.T., Twiss, J.L., Beattie, C.E., Steen, J.A. and Sahin, M. Interaction of survival of motor neuron (SMN) and HuD proteins with mRNA cpg15 rescues motor neuron axonal deficits. Proc. Natl. Acad. Sci. USA 108 (2011) 10337-10342.Google Scholar

  • 9. Kos, K., Fine, L. and Coulombe, J.N. Activin type II receptors in embryonic dorsal root ganglion 351 neurons of the chicken. J. Neurobiol. 47 (2001) 93-108.Google Scholar

  • 10. Hou, S., Tian, W., Xu, Q., Cui, F., Zhang, J., Lu, Q. and Zhao, C. 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. Neuroscience 137 (2006) 519-529Google Scholar

  • 11. Jacquez, P., Lei, N., Weigt, D., Xiao, C. and Sun, J. 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. Protein Expr. Purif. 95 (2014) 149-155.Google Scholar

  • 12. Gagliardo, B., Faye, A., Jaouen, M., Deschemin, J.C., Canonne-Hergaux, F., Vaulont, S. and Sari, M.A. Production of biologically active forms of recombinant hepcidin, the ironregulatory hormone. FEBS J. 275 (2008) 3793-3803.Google Scholar

  • 13. Mahmoudi, S., Abtahi, H., Bahador, A., Mosayebi, G., Salmanian, A.H. and Teymuri, M. Optimizing of nutrients for high level expression of recombinant streptokinase using pET32a expression system. Maedica (Buchar) 7 (2012) 241-246.Google Scholar

  • 14. Picard, N., Leslie, J.H., Trowbridge, S.K., Subramanian, J., Nedivi, E. and Fagiolini, M. Aberrant development and plasticity of excitatory visual cortical networks in the absence of cpg15. J. Neurosci. 34 (2014) 3517-3522.CrossrefGoogle Scholar

  • 15. Sato, H., Fukutani, Y., Yamamoto, Y., Tatara, E., Takemoto, M., Shimamura, K. and Yamamoto, N. Thalamus-derived molecules promote survival and dendritic growth of developing cortical neurons. J. Neurosci. 32 (2012) 15388-15402.Web of ScienceCrossrefGoogle Scholar

  • 16. Han, D., Qin, B., Liu, G., Liu, T., Ji, G., Wu, Y. and Yu, L. Characterization of neuritin as a novel angiogenic factor. Biochem. Biophys. Res. Commun. 415 (2011) 608-612.Web of ScienceGoogle Scholar

  • 17. MacLusky, N.J., Hajszan, T., Prange-Kiel, J. and Leranth, C. Androgen modulation of hippocampal synaptic plasticity. Neuroscience 138 (2006) 957-965.Google Scholar

  • 18. Sharma, T.P., Liu, Y., Wordinger, R.J., Pang, I.H. and Clark, A.F. Neuritin 1 promotes retinal ganglion cell survival and axonal regeneration following optic nerve crush. Cell Death Dis. 6 (2015) e1661. DOI:10.1038/cddis.2015.22.CrossrefGoogle Scholar

  • 19. Cappelletti, G., Galbiati, M., Ronchi, C., Maggioni, M.G., Onesto, E. and Poletti, A. Neuritin (cpg15) enhances the differentiating effect of NGF on neuronal PC12 cells. J. Neurosci. Res. 85 (2007) 2702-2713.Google Scholar

  • 20. Karamoysoyli, E., Burrnand, R.C. and Tomlinson, D.R. Neuritin mediates nerve growth factor-induced axonal regeneration and is deficient in experimental diabetic neuropathy. Diabetes 57 (2008) 181-189.Web of ScienceGoogle Scholar

  • 21. Msika, O., Brand, A., Crawford, M.A. and Yavin, E. NGF blocks polyunsaturated fatty acids biosynthesis in n-3 fatty acid-supplemented PC12 cells. Biochim. Biophys. Acta 1821 (2012) 1022-1030.Web of ScienceGoogle Scholar

  • 22. Zhang, Y., Zhang, S., Xian, L., Tang, J., Zhu, J., Cui, L., Li, S., Yang, L. and Huang, J. Expression and purication of recombinant human neuritin from Pichia pastoris and a partial analysis of itsneurobiological activity in vitro. Appl. Microbiol. Biotechnol. 99 (2015) 8035-8043.Google Scholar

  • 23. Zhou, S. and Zhou, J. Neuritin, a neurotrophic factor in nervous system physiology. Curr. Med. Chem. 21 (2014) 1212-1219. CrossrefWeb of ScienceGoogle Scholar

About the article

Received: 2015-10-06

Accepted: 2015-12-31

Published Online: 2016-03-05

Published in Print: 2015-12-01


Citation Information: Cellular and Molecular Biology Letters, Volume 20, Issue 5, Pages 965–973, ISSN (Online) 1689-1392, DOI: https://doi.org/10.1515/cmble-2015-0057.

Export Citation

© University of Wroclaw, Poland. Copyright Clearance Center

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[1]
Shuai Zhang, Yanhong Huang, Jingling Zhu, Liya Shan, Jianfeng Gao, Yunhua Zhang, Na Yu, Lei Yang, and Jin Huang
Gene, 2018

Comments (0)

Please log in or register to comment.
Log in