Preparation of recombinant neuritin protein

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-07-11 DOI:10.1016/j.pep.2024.106554

Pingping Meng , Liyan Zhu , Jiatong Guo , Yuanyuan Li , Yu Wei , Jiawei Sun , Jingling Zhu

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Abstract

Neuritin plays an important role in promoting nerve injury repair and maintaining synaptic plasticity, making it a potential therapeutic target for the treatment of nerve injury and neurodegenerative diseases. The present study aimed to obtain an active, unlabeled neuritin protein. Initially, a neuritin protein expression system with an enterokinase site was constructed in Escherichia coli. After optimizing induction conditions and screening for high expression, a neuritin recombinant protein with purity exceeding 85 % was obtained through Ni-affinity chromatography. Subsequently, unlabeled neuritin with a molecular weight of 11 kDa was obtained through the enzymatic cleavage of the His label using an enterokinase. Furthermore, a neuritin recombinant protein with purity exceeding 95 % was obtained using gel chromatography. Functional investigations revealed that neurite outgrowth of PC12 cells was stimulated by the isolated neuritin. This study establishes a method to obtain active and unlabeled neuritin protein, providing a foundation for subsequent research on its biological functions.

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制备重组神经肽蛋白。

神经营养素在促进神经损伤修复和维持突触可塑性方面发挥着重要作用，是治疗神经损伤和神经退行性疾病的潜在治疗靶点。本研究旨在获得活性的、未标记的神经肽蛋白。首先，在大肠杆菌中构建了带有肠激酶位点的神经肽表达系统。经过优化诱导条件和高表达筛选，通过镍亲和层析获得了纯度超过 85% 的神经肽重组蛋白。随后，利用肠激酶酶解 His 标记，得到分子量为 11 kDa 的未标记神经肽。此外，利用凝胶色谱法还获得了纯度超过 95% 的神经肽重组蛋白。功能研究发现，分离出的神经肽能刺激 PC12 细胞的神经元突起。这项研究建立了一种获得活性和未标记的神经肽蛋白的方法，为后续研究其生物功能奠定了基础。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.