Fusion with CTP increases the stability of recombinant neuritin.
Carboxyl-terminal peptide
Chinese hamster ovary cells
Neuritin
Stability
Journal
Protein expression and purification
ISSN: 1096-0279
Titre abrégé: Protein Expr Purif
Pays: United States
ID NLM: 9101496
Informations de publication
Date de publication:
12 2023
12 2023
Historique:
received:
10
06
2023
revised:
25
07
2023
accepted:
08
08
2023
medline:
18
9
2023
pubmed:
12
8
2023
entrez:
11
8
2023
Statut:
ppublish
Résumé
Neuritin is a vital neurotrophin that plays an essential role in recovery from nerve injury and neurodegenerative diseases and may become a new target for treating these conditions. However, improving neuritin protein stability is an urgent problem. In this study, to obtain active and stable neuritin proteins, we added a carboxyl-terminal peptide (CTP) sequence containing four O-linked glycosylation sites to the C-terminus of neuritin and cloned it into the Chinese hamster ovary (CHO) expression system. The neuritin-CTP protein was purified using a His-Tag purification strategy after G418 screening of stable high-expression cell lines. Ultimately, we obtained neuritin-CTP protein with a purity >90%. Functional analyses showed that the purified neuritin-CTP protein promoted the neurite outgrowth of PC12 cells, and stability experiments showed that neuritin stability was increased by adding CTP. These results indicate that neuritin protein-CTP fusion effectively increases stability without affecting secretion and activity. This study offers a sound strategy for improving the stability of neuritin protein and provides material conditions for further study of the function of neuritin.
Identifiants
pubmed: 37567400
pii: S1046-5928(23)00115-8
doi: 10.1016/j.pep.2023.106344
pii:
doi:
Substances chimiques
GPI-Linked Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
106344Informations de copyright
Copyright © 2023 Elsevier Inc. All rights reserved.