Effects of Different Domains on Activity of Wheat (Triticum aestivum) TaNRX1-D
CHENG Jie1,*, SONG Tian-Qi1,*, WEI Fan1, LI Rui-Bo1, YU Ming1, HAO Xi-Ying1, TIAN Shu-Yuan1, QIN Wu-Sa1, YANG Song-Jie2,**, ZHANG Xiao-Ke1,**
1 College of Agronomy, Northwest A&F University, Yangling 712100, China;
2 School of Modern Agriculture & Biotechnology, Ankang University, Ankang 725000, China;
Abstract：Thioredoxin (TRX) contains highly conserved typical domains with oxidative active site WCG/PPC, and is a class of small molecular multifunctional proteins widely existing in organisms. Nucleoredoxin (NRX), a member of the TRX superfamily, was contained multiple domains typical and atypical of TRX, and has the potential of redox activity. According to bioinformatics analysis, the Triticum aestivum TaNRX1-D contains 3 TRX domains: domain 1 contained WCPPC, domain 2 contained GYPPV and domain3 contained WCGPC, while the domain 2 did not harbour such an activity site. In order to further explore the different structural domain influence on TaNRX1-D protein activity, this experiment truncated the protein sequence at different lengths according to the predicted physicochemical properties of the TaNRX1-D domain protein. The corresponding protein products were obtained by constructing prokaryotic expression vectors and purified, respectively. TaNRX1-D possesses TRX activity in vitro. Compared with 1.0 µmol/L or 2.5 µmol/L, 5.0 µmol/L TaNRX1-D displayed high insulin reduction activity in a dose-dependent manner. Among the 5 domains of TaNRX1-D, the insulin reduction activity of the 5 domains as follows: NRX-D3 (NRX-domain3)>NRX-D1 (NRX-domain1)>NRX-D2 (NRX-domain2), and each of the 5 domains alone was less active than the NRX-DQ protein. The analysis of three structural domains inside the protein on the reducing activity of TaNRX1-D protein will provide a new reference for further understanding of the function of TaNRX1-D protein.
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