caa a22 4500 605498679 CHVBK 20210128100547.0 cr unu---uuuuu 210128e20151001xx s 000 0 eng 10.1007/s00253-015-6649-3 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6649-3 Expression and purification of recombinant human neuritin from Pichia pastoris and a partial analysis of its neurobiological activity in vitro [Elektronische Daten] [Yunhua Zhang, Shujun Zhang, Lingling Xian, Juan Tang, Jingling Zhu, Lijuan Cui, Shanman Li, Lei Yang, Jin Huang] 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. Springer-Verlag Berlin Heidelberg, 2015 Dorsal root ganglion (DRG) nationallicence Neuritin nationallicence PC12 cells nationallicence Pichia pastoris nationallicence Purification nationallicence Zhang Yunhua The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Zhang Shujun The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Xian Lingling The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Tang Juan The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Zhu Jingling The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Cui Lijuan The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Li Shanman Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, 832003, Shihezi, Xinjiang, China aut Yang Lei School of Medicine, Hangzhou Normal University, 310036, Hangzhou, Zhejiang, China aut Huang Jin The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8035-8043 0175-7598 99:19<8035 2015 99 253 https://doi.org/10.1007/s00253-015-6649-3 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6649-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Yunhua The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Zhang Shujun The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Xian Lingling The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Tang Juan The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Zhu Jingling The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Cui Lijuan The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Li Shanman Key Laboratory for Green Processing of Chemical Engineering of Xinjiang Bingtuan, School of Chemistry and Chemical Engineering, Shihezi University, 832003, Shihezi, Xinjiang, China aut NATIONALLICENCE 700 1- Yang Lei School of Medicine, Hangzhou Normal University, 310036, Hangzhou, Zhejiang, China aut NATIONALLICENCE 700 1- Huang Jin The Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Department of Biochemistry, School of Medicine, Shihezi University, 832002, Shihezi, Xinjiang, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/19(2015-10-01), 8035-8043 0175-7598 99:19<8035 2015 99 253