A novel 76-mer peptide mimic with the synergism of superoxide dismutase and glutathione peroxidase
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The balance of oxidation and reduction in the body requires the synergistic effect of various antioxidant enzymes. Therefore, the construction of enzyme mimics with multiple antioxidant activities is important and beneficial for further research on the synergistic effects of antioxidant enzymes and their mechanism of action. To explore the synergistic effect of superoxide dismutase (SOD) and glutathione peroxidase (GPx), a 76-mer selenium-containing peptide (Se-76P) mimic containing the active SOD and GPx centers was designed. Moreover, a cell-penetrating peptide was introduced into Se-76P by structure modeling, and then, Se-76P was expressed by a single-protein production combined with the cysteine auxotrophic double-expression system of Escherichia coli. The results suggest that Se-76P exhibits SOD and GPx activities, following the GPx activity of 109 U/mg protein and the SOD activity of 1218 U/mg protein. The labeled Se-76P with FITC fluorescence was verified to enter the L02 cells successfully; it improved the antioxidant activity in cells and promoted the consumption of glucose and synthesis of glycogen. The injection of Se-76P subcutaneously decreased the levels of blood glucose and malondialdehyde of lipid peroxidation produced in mice, indicating that Se-76P had antioxidative properties and a certain regulatory role of glucose metabolism. The data analysis provides further clarification that Se-76P can regulate insulin signal transduction to play an insulin-like role, which not only has a greater significance for further elucidating the catalytic mechanism of the enzyme and their synergistic effects on each other but also has enormous medicinal potential.
KeywordsEnzyme mimics Superoxide dismutase Glutathione peroxidase Single-protein production Antioxidant activity
We thank Prof. August Bőck (Lehrstuhl für Mikrobiologie der Universität München, Germany) and Dr. Marie-Paule Strub (Laboratory of Molecular Biophysics, National Heart, Lung, and Blood Institute, National Institutes of Health, USA) for providing strain BL21 (DE3) selB::kan cys51E (BL21cysE51). We also thank Prof. Mu Ying (Research Center for Analytical Instrumentation, Zhejiang University, Hangzhou, China) for providing E. coli DH5α and L02 hepatocytes.
- Li XW, Su X, Zhang Y, Sun YP, Zhao Y, Zhai Y, Wang QY (2015) An improved calcium chloride method preparation and transformation of competent cells. Afr J Biotechnol 9(50):8549–8554Google Scholar
- Peter K, Graeber J, Kipriyanov S, Zewe-Welschof M, Runge MS, Kübler W, Little M, Bode C (2000) Construction and functional evaluation of a single-chain antibody fusion protein with fibrin targeting and thrombin inhibition after activation by factor Xa. Circulation 101:1158–1164CrossRefPubMedGoogle Scholar
- Rayman MP, Thompson AJ, Bekaert B, Catterick J, Galassini R, Hall E, Warren-Perry M, Beckett GJ (2008) Randomized controlled trial of the effect of selenium supplementation on thyroid function in the elderly in the United Kingdom. The American Journal of Clinical Nutrition 87 (2):370–378Google Scholar
- Wang C, Wan P, Gong PS, Lv LM, Xu YW, Zhao Y, He B, Zhao G, Yan GL, Mu Y, Lv SW, Luo GM (2011) Novel selenium-containing human single-chain variable fragment with glutathione peroxidase activity from computer-aided molecular design. CHEM RES CHINESE UNIVERSITIES 27(5):813–819Google Scholar