Active Site Mimicry of Glutathione Peroxidase by Glutathione Imprinted Selenium-Containing Trypsin
AbstractIn order to overcome the instability of natural glutathione peroxidase (GPx), scientists endeavor to produce GPx mimics. The popular method first uses biological imprinting (BI) to produce the substrate binding sites and then employs chemical mutation (CM) to obtain the catalytic site. However, BICM has a drawback in that the catalytic site is not clear. Some researchers therefore tried to change the order of the method. These new GPx mimics were prepared by first producing the catalytic site through chemical mutation, and then employing biological imprinting to produce the substrate binding sites (CMBI). It has a clear catalytic site, but its determination of enzyme activity and kinetic analysis are still not elucidated. In this study, we used CMBI to synthesize a GPx mimic using trypsin as the imprinted molecule and GSSG as the template molecule and compared the enzyme activity of the four intermediates (Trypsin-SeO2H (TSeO2H), Trypsin-Se-SG (TSeSG), Imprinted Trypsin-Se-SG (ITSeSG), Cross-linked Imprinted Trypsin-Se-SG (CITSeSG), we analyzed the properties of intermediate products. All values are the means of at least four determinations, ITSeSG was produced from TSeSG through bio-imprinting, the activity of GPx mimics synthesized by CMBI was 5.7 times greater than native GPx, because of bio-imprinting make KmGSH value of the mimics decreased from 4.82 ± 0.27 mM (TSeSG) to 0.52 ± 0.05 mM (ITSeSG). This proves that bio-imprinting is the reason for increased substrate binding capability. View Full-Text
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Huang, Y.; Ge, D.-Y.; Zong, H.; Yin, J.-X.; Qu, X.-N.; Lv, S.-W. Active Site Mimicry of Glutathione Peroxidase by Glutathione Imprinted Selenium-Containing Trypsin. Catalysts 2017, 7, 282.
Huang Y, Ge D-Y, Zong H, Yin J-X, Qu X-N, Lv S-W. Active Site Mimicry of Glutathione Peroxidase by Glutathione Imprinted Selenium-Containing Trypsin. Catalysts. 2017; 7(10):282.Chicago/Turabian Style
Huang, Yi; Ge, Dan-Yang; Zong, Hui; Yin, Ju-Xin; Qu, Xiao-Nan; Lv, Shao-Wu. 2017. "Active Site Mimicry of Glutathione Peroxidase by Glutathione Imprinted Selenium-Containing Trypsin." Catalysts 7, no. 10: 282.
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