Expression and purification of recombinant human neuritin from Pichia pastoris and a partial analysis of its neurobiological activity in vitro

Verfasser / Beitragende:
[Yunhua Zhang, Shujun Zhang, Lingling Xian, Juan Tang, Jingling Zhu, Lijuan Cui, Shanman Li, Lei Yang, Jin Huang]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/19(2015-10-01), 8035-8043
Format:
Artikel (online)
ID: 605498679
LEADER caa a22 4500
001 605498679
003 CHVBK
005 20210128100547.0
007 cr unu---uuuuu
008 210128e20151001xx s 000 0 eng
024 7 0 |a 10.1007/s00253-015-6649-3  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6649-3 
245 0 0 |a Expression and purification of recombinant human neuritin from Pichia pastoris and a partial analysis of its neurobiological activity in vitro  |h [Elektronische Daten]  |c [Yunhua Zhang, Shujun Zhang, Lingling Xian, Juan Tang, Jingling Zhu, Lijuan Cui, Shanman Li, Lei Yang, Jin Huang] 
520 3 |a Neuritin (also known as candidate plasticity gene 15 (cpg15)) is a neurotrophic factor that was recently discovered in a screen aimed at identifying genes involved in activity-dependent synaptic plasticity. Neuritin plays multiple roles in both neural development (Chen et al. Zhonghua Yan Ke Za Zhi 46:978-983 2010; Corriveau et al. J Neurosci 19:7999-8008 1999; Lee and Nedivi J Neurosci 22:1807-1815 2002) and synaptic plasticity (Fujino et al. Gene Dev 25:2674-2685 2011; Leslie and Nedivi Prog 14 Neurobiol 94:223-237 2011; Loebrich and Nedivi Physiol Rev 89:1079 2009). In this study, to produce bioactive, soluble recombinant human neuritin protein, a portion of NRN1 was cloned into the Pichia pastoris expression vector pPIC9K. The recombinant vector was then transformed into the methylotrophic yeast strain P. pastoris GS115, and a shaking flask method and His-tag purification strategy were utilized to express and purify neuritin protein. The resulting protein had a molecular mass of approximately 11kDa, and subsequent functional analyses indicated that the purified neuritin promoted neurite outgrowth from embryonic chicken dorsal root ganglions, while also prolonging the survival of these ganglions, and from PC12 cells. These findings suggest that neuritin was expressed effectively in vitro and that this protein may play a role in stimulating neurite outgrowth of both dorsal root ganglions and PC12 cells. This study provides a novel strategy for the large-scale production of bioactive neuritin, which will enable further study of the biological function of this protein. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Dorsal root ganglion (DRG)  |2 nationallicence 
690 7 |a Neuritin  |2 nationallicence 
690 7 |a PC12 cells  |2 nationallicence 
690 7 |a Pichia pastoris  |2 nationallicence 
690 7 |a Purification  |2 nationallicence 
700 1 |a Zhang  |D Yunhua  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Zhang  |D Shujun  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Xian  |D Lingling  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Tang  |D Juan  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Zhu  |D Jingling  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Cui  |D Lijuan  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Li  |D Shanman  |u Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, 832003, Shihezi, Xinjiang, China  |4 aut 
700 1 |a Yang  |D Lei  |u School of Medicine, Hangzhou Normal University, 310036, Hangzhou, Zhejiang, China  |4 aut 
700 1 |a Huang  |D Jin  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/19(2015-10-01), 8035-8043  |x 0175-7598  |q 99:19<8035  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6649-3  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6649-3  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhang  |D Yunhua  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhang  |D Shujun  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Xian  |D Lingling  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Tang  |D Juan  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Zhu  |D Jingling  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Cui  |D Lijuan  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Li  |D Shanman  |u Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, 832003, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Yang  |D Lei  |u School of Medicine, Hangzhou Normal University, 310036, Hangzhou, Zhejiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Huang  |D Jin  |u The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/19(2015-10-01), 8035-8043  |x 0175-7598  |q 99:19<8035  |1 2015  |2 99  |o 253