毕赤酵母糖基化对布鲁菌P39抗原诱导巨噬细胞吞噬的影响

doi:10.13560/j.cnki.biotech.bull.1985.2018-0170

生物技术通报 ›› 2018, Vol. 34 ›› Issue (9): 237-243.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0170

毕赤酵母糖基化对布鲁菌P39抗原诱导巨噬细胞吞噬的影响

刘思阳, 王岩, 付玉芹, 胡祥坤, 王茗宇, 卢天成, 王秀然

吉林农业大学生命科学学院生物反应器与药物开发教育部工程研究中心,长春130118

收稿日期:2018-02-27 出版日期:2018-09-26 发布日期:2018-10-10
作者简介:刘思阳 ,女,硕士研究生,研究方向：微生物资源开发与利用方面;E-mail： siyang5676@163.com
基金资助:
国家自然科学基金项目（31302062）,吉林省科技支撑项目（20160312008ZG）,吉林省自然基金项目（20180101260JC）

Effect of Glycosylation of Brucella P39 Antigen in Pichia pastoris on Macrophage Phagocytosis

LIU Si-yang, WANG Yan, FU Yu-qin, HU Xiang-kun, WANG Ming-yu, LU Tian-cheng, WANG Xiu-ran

Bioreactor and Drug Development Engineering Research Center of the Ministry of Education,College of Life Science,Jilin Agricultural University,Changchun 130118

Received:2018-02-27 Published:2018-09-26 Online:2018-10-10

摘要/Abstract

摘要： P39蛋白是通过蛋白组学从布鲁菌中鉴定出的具有较高免疫原性的优势抗原蛋白之一。本研究对布鲁菌的优势抗原蛋白P39基因在毕赤酵母中进行表达,并对其进行纯化及Western blot鉴定,利用核磁共振接合PNGaseF糖苷酶进行蛋白糖基化鉴定,并研究其与巨噬细胞相互作用的速度。结果表明,P39蛋白在毕赤酵母GS115中呈分泌表达,表达量可达5.05 g/L。核磁共振结果表明,重组P39蛋白被糖基化修饰。修饰后的蛋白被巨噬细胞捕获效应明显强于原核蛋白,说明糖基化修饰可能影响目标蛋白与细胞相互作用的效率,为进一步研究糖基化对P39蛋白的影响及布鲁菌糖基化疫苗候选抗原的研究提供参考。

关键词: 布鲁菌, P39蛋白, 毕赤酵母, 糖基化

Abstract: P39 protein is one of the most predominant antigenic proteins identified from Brucella by proteomics. First,the dominant antigen protein gene P39 was expressed in Pichia pastoris,and the expressed protein was purified and identified by Western blot. Then,the glycosylation of the protein was identified by NMR（nuclear magnetic resonance）conjugated PNGaseF glycosylase,and the interaction speed between the protein and macrophage cell was studied. The results showed that P39 protein was secreted in P. pastoris GS115 and the expression level was up to 5.05 g/L. NMR results demonstrated that the recombinant P39 protein was modified by glycosylation. The macrophage capture effect of the modified protein was significantly stronger than prokaryotic protein,indicating that the glycosylation modification may affect the efficiency of the interaction between the target protein and the cell,which provides a reference for further exploring the effect of glycosylation on P39 protein and candidate antigen of Brucella glycosylated vaccine.

Key words: Brucella, P39 protein, Pichia pastoris, glycosylation

刘思阳, 王岩, 付玉芹, 胡祥坤, 王茗宇, 卢天成, 王秀然. 毕赤酵母糖基化对布鲁菌P39抗原诱导巨噬细胞吞噬的影响[J]. 生物技术通报, 2018, 34(9): 237-243.

LIU Si-yang, WANG Yan, FU Yu-qin, HU Xiang-kun, WANG Ming-yu, LU Tian-cheng, WANG Xiu-ran. Effect of Glycosylation of Brucella P39 Antigen in Pichia pastoris on Macrophage Phagocytosis[J]. Biotechnology Bulletin, 2018, 34(9): 237-243.

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毕赤酵母糖基化对布鲁菌P39抗原诱导巨噬细胞吞噬的影响

Effect of Glycosylation of Brucella P39 Antigen in Pichia pastoris on Macrophage Phagocytosis

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