Effects of Domains of Wheat Protein Disulfide Isomerase on its Properties

被引：1
|
作者：
Hu S.-Q. [1 ]
Huang Z. [1 ]
Liu G. [1 ,2 ]
Huang Y.-B. [3 ]
Li L. [1 ]
Hou Y. [3 ]
机构：
[1] School of Food Science and Engineering, South China University of Technology, Guangzhou, 510640, Guangdong
[2] Sericultural and Agri-Food Research Institute, GAAS, Guangzhou, 510610, Guangdong
[3] School of Light Industry Science and Engineering, South China University of Technology, Guangzhou, 510640, Guangdong
来源：
| 1600年 / South China University of Technology卷 / 45期
基金：
中国国家自然科学基金;
关键词：
Active-site; Domain; Protein disulfide isomerase; Truncated protein; Wheat;
D O I：
10.3969/j.issn.1000-565X.2017.11.013
中图分类号：
学科分类号：
摘要：
The wheat protein disulfide isomerase (wPDI) contains four thioredoxin domains a-b-b'-a' and a C-terminal tail c. In order to investigate the effect of each domain of wPDI on its properties, eight truncated proteins of wPDI containing different domain combinations were constructed by subcloning. The target proteins were prepared after expression and purification, and then their enzymatic properties and products of protein electrophoresis were determined and analyzed. The results indicate that eight truncated proteins of wPDI are expressed in E.coli BL21, and that, after the metal chelate affinity chromatography and the size exclusion chromatography, the truncated proteins of wPDI of high purity are obtained. The results from activities assay indicate that all domains of wPDI contribute to its disulfide bond oxidoreductase activity and molecular chaperone activity, and that, the truncation of tail c has no effect on its isomerase activity but retains the critical amino acid binding site for the molecular chaperone activity. Moreover, the electrophoretic analysis of the truanted proteins A and A' demonstrates that the active-site cysteines in domain a are primarily in an oxidized state while those of domain a' are primarily in a reduced state. Therefore, these results lay a foundation for deeply understanding the function and property of wPDI. © 2017, Editorial Department, Journal of South China University of Technology. All right reserved.
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页码：92 / 99
页数：7
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