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A novel 76-mer peptide mimic with the synergism of superoxide dismutase and glutathione peroxidase

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Abstract

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.

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References

  • Ahmed HH, El-Maksoud MDA, Moneim Ahmed EA, Aglan HA (2017) Pre-clinical study for the antidiabetic potential of selenium nanoparticles. Biol Trace Elem Res 177(2):267–280

    Article  CAS  PubMed  Google Scholar 

  • Andreyev AY, Kushnareva YE, Starkov AA (2005) Mitochondrial metabolism of reactive oxygen species. Biochem Mosc 70(2):200–214

    Article  CAS  Google Scholar 

  • Antonyuk SV, Strange RW, Marklund SL, Hasnain SS (2009) The structure of human extracellular copper–zinc superoxide dismutase at 1.7 Å resolution: insights into heparin and collagen binding. J Mol Biol 388(2):310–326

    Article  CAS  PubMed  Google Scholar 

  • Atif F, Yousuf S, Agrawal SK (2008) Restraint stress-induced oxidative damage and its amelioration with selenium. Eur J Pharmacol 600(1–3):59–63

    Article  CAS  PubMed  Google Scholar 

  • Awasthi YC, Dao DD, La AK, Srivastava SK (1979) Purification and properties of glutathione peroxidase from human placenta. Biochem J 177(2):471–476

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254

    Article  CAS  PubMed  Google Scholar 

  • Casi G, Roelfes G, Hilvert D (2008) Selenoglutaredoxin as a glutathione peroxidase mimic. Chembiochem 9:1623–1631

    Article  CAS  PubMed  Google Scholar 

  • Dai C, Tang SS, Li DW, Zhao KN, Xiao XL (2015) Curcumin attenuates quinocetone-induced oxidative stress and genotoxicity in human hepatocyte L02 cells. Toxicol Mech Methods 25(4):340–346

    Article  CAS  PubMed  Google Scholar 

  • Guidotti G, Brambilla L, Rossi D (2017) Cell-penetrating peptides: from basic research to clinics. Trends Pharmacol Sci 38(4):406–424

    Article  CAS  PubMed  Google Scholar 

  • Hungerford G, Benesch J, Mano JF, Reis RL (2007) Effect of the labelling ratio on the photophysics of fluorescein isothiocyanate (FITC) conjugated to bovine serum albumin. Photochemical & Photobiological Sciences 6(2):152–158

    Article  CAS  Google Scholar 

  • Ibuki FK, Simões A, Nicolau J, Nogueira FN (2013) Laser irradiation affects enzymatic antioxidant system of streptozotocin-induced diabetic rats. Lasers Med Sci 28(3):911–918

    Article  PubMed  Google Scholar 

  • Ismail TA, Nassan MA, Alkhedaide AQ, Soliman MM (2016) Modulation of aquaporins expressions and its impact on alleviating diabetic complications in rats: role of Citrullus colocynthis. National Journal of Physiology, Pharmacy and Pharmacology 6(4):310–322

    Article  CAS  Google Scholar 

  • Lang SH, Spratt DE, Guillemette JG, Palmer M (2005) Dual-targeted labeling of proteins using cysteine and selenomethionine residues. Anal Biochem 342:271–279

    Article  CAS  PubMed  Google Scholar 

  • Li GL, Ye Y, Kang JJ, Yao XY, Zhang YZ, Jiang W, Gao M, Dai YD, Xin YQ, Wang Q, Yin ZM, Luo L (2012) L-Theanine prevents alcoholic liver injury through enhancing the antioxidant capability of hepatocytes. Food Chem Toxicol 50:363–372

    Article  CAS  PubMed  Google Scholar 

  • 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):8549–8554

    Google Scholar 

  • Lippman SM, Klein EA, Goodman PJ, Lucia MS, Thompson IM, Ford LG, Parnes HL, Minasian LM, Gaziano JM, Hartline JA, Parsons JK, Bearden JD III (2009) Effect of selenium and vitamin E on risk of prostate cancer and other cancers. JAMA 301(1):39–51

    Article  CAS  PubMed  Google Scholar 

  • Mao LL, Stathopulos PB, Ikura M, Inouye M (2010) Secretion of human superoxide dismutase in Escherichia coli using the condensed single-protein-production system. Protein Sci 19:2330–2335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marklund SL (1982) Human copper-containing superoxide dismutase of high molecular weight. Proc Natl Acad Sci U S A 79(24):7634–7638

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Müller S, Heider J, Böck A (1997) The path of unspecific incorporation of selenium in Escherichia coli. Arch Microbiol 168(5):421–427

    Article  PubMed  Google Scholar 

  • Muller S, Senn H, Gsel B, Vetter W, Baron C, Böck A (1994) The formation of diselenide bridges in proteins by incorporation of selenocysteine residues: biosynthesis and characterization of (Se)2-thioredoxin. Biochemistry 33:3404–3412

    Article  CAS  PubMed  Google Scholar 

  • Murata M, Yoshida Y, Osono M, Ohashi N, Oyanagi M, Urakami H, Tamura A, Nogami S, Tanaka H, Kawamura A (1986) Production and characterization of monoclonal strain-specific antibodies against prototype strains of Rickettsia tsutsugamushi. Microbiol Immunol 30(7):599–610

    Article  CAS  PubMed  Google 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 101:1158–1164

    Article  CAS  PubMed  Google Scholar 

  • Ramière C, Rodriguez J, Enache LS, Lotteau V, André P, Diaza O (2014) Activity of hexokinase is increased by its interaction with hepatitis C virus protein NS5A. J Virol 88(6):3246–3254

    Article  PubMed  PubMed Central  Google 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):370–378

  • Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Selenium: biochemical role as a component of glutathione peroxidase. Science 179:588–590

    Article  CAS  PubMed  Google Scholar 

  • Sanchez JF, Hoh F, Strub MP, Aumelas A, Dumas C (2002) Structure of the cathelicidin motif of protegrin-3 precursor: structural insights into the activation mechanism of an antimicrobial protein. Structure 10:1363–1370

    Article  CAS  PubMed  Google Scholar 

  • Schägger H (2006) Tricine-SDS-PAGE. Nat Protoc 1(1):16–22

    Article  PubMed  Google Scholar 

  • Simonyan L, Légiot A, Lascu I, Durand G, Giraud MF, Gonzalez C, Manon S (2017) The substitution of Proline 168 favors Bax oligomerization and stimulates its interaction with LUVs and mitochondria. Biochim Biophys Acta 1859:1144–1155

    Article  CAS  PubMed  Google Scholar 

  • Staneviciene I, Ivanov L, Kursvietiene L, Viezeliene D (2017) Short-term effects of aluminum and selenium on redox status in mice brain and blood. Trace Elements and Electrolytes 34(2):74–80

    Article  Google Scholar 

  • Stranges S, Sieri S, Vinceti M, Grioni S, Guallar E, Laclaustra M, Muti P, Berrino F, Krogh V (2010) A prospective study of dietary selenium intake and risk of type 2 diabetes. BMC Public Health 10:564–572

    Article  PubMed  PubMed Central  Google Scholar 

  • Strub MP, Hoh F, Sanchez JF, Strub JM, Böck A, Aumelas A, Dumas C (2003) Selenomethionine and selenocysteine double labeling strategy for crystallographic phasing. Structure 11:1359–1367

    Article  CAS  PubMed  Google Scholar 

  • Stein V, Alexandrov K (2015) Synthetic protein switches: design principles and applications. Trends Biotechnol 33(2):101–110

    Article  CAS  PubMed  Google Scholar 

  • Suzuki M, Roy R, Zheng H, Woychik N, Inouye M (2006) Bacterial bioreactors for high yield production of recombinant protein. J Biol Chem 281(49):37559–37565

    Article  CAS  PubMed  Google Scholar 

  • Suzuki M, Zhang J, Liu M, Woychik NA, Inouye M (2005) Single protein production in living cells facilitated by an mRNA interferase. Mol Cell 18:253–261

    Article  CAS  PubMed  Google Scholar 

  • Tainer JA, Getzoff ED, Richardson JS, Richardson DC (1983) Structure and mechanism of copper, zinc superoxide dismutase. Nature 306:284–287

    Article  CAS  PubMed  Google 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):813–819

    CAS  Google Scholar 

  • Wang H, Xue Z, Wang Q, Feng X, Shen Z (2008) Propofol protects hepatic L02 cells from hydrogen peroxide-induced apoptosis via activation of extracellular signal-regulated kinases pathway. Anesth Analg 107(2):534–540

    Article  CAS  PubMed  Google Scholar 

  • Wang XH, Jw Y, Lei XG (2014) Glutathione peroxidase mimic ebselen improves glucose-stimulated insulin secretion in murine islets. Antioxid Redox Signal 20(2):191–203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang ZL, Yu WD, Chen SC (1985) Frequency of erythrocyte pyruvate kinase deficiency in Chinese infants. Am J Hematol 20(2):139–144

    Article  Google Scholar 

  • Yan F, Yan GL, Lv S, Shen N, Mu Y, Chen T, Gong PS, Xu YW, Lv LM, Liu JQ, Shen JC, Luo GM (2011) A novel 65-mer peptide imitates the synergism of superoxide dismutase and glutathione peroxidase. Int J Biochem Cell Biol 43:1802–1811

    Article  CAS  PubMed  Google Scholar 

  • Yan F, Yang WK, Li XY, Lin TT, Lun YN, Lin F, Lv SW, Yan GL, Liu JQ, Shen JC, Mu Y, Luo GM (2008) A trifunctional enzyme with glutathione S-transferase, glutathione peroxidase and superoxide dismutase activity. BBA-Gen Subjects 1780:869–872

    Article  CAS  Google Scholar 

  • Yu HJ, Ge Y, Wang Y, Lin CT, Li J, Liu X, Zang TZ, Xu JQ, Liu JQ, Luo GM, Shen JC (2007) A fused selenium-containing protein with both GPx and SOD activities. Biochem Biophys Res Commun 358:873–878

    Article  CAS  PubMed  Google Scholar 

  • Zhang Y, Zhang J, Hoeflich KP, Ikura M, Qing G, Inouye M (2003) MazF cleaves cellular mRNAs specifically at ACA to block protein synthesis in Escherichia coli. Mol Cell 12:913–923

    Article  CAS  PubMed  Google Scholar 

  • Zinoni F, Birkmann A, Leinfelder W, Böck A (1987) Cotranslational insertion of selenocysteine into formate dehydrogenase from Escherichia coli directed by a UGA codon. Proc Natl Acad Sci 84(10):3156–3160

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zou XF, Ji YT, Gao G, Zhu XJ, Lv SW, Yan F, Han SP, Chen X, Gao CC, Liu JQ, Luo GM (2010) A novel selenium- and copper-containing peptide with both superoxide dismutase and glutathione peroxidase activities. J Microbiol Biotechnol 20:88–93

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

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.

Funding

This work was supported by the Jilin Science and Technology Support Program (20150101131JC) and the Jilin Agricultural Science and Technology College Basic Scientific Research Operation Cost fund (2014 Z01).

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Authors and Affiliations

  1. College of Bioengineering, Jilin Agricultural Science and Technology University, Jilin City, Jilin Province, 132101, China

    Yawei Xu, Yan Zhou, Rui Yin, Haijiao Chu & Junling Wang

  2. School of Basic Medicine, Jilin City, Jilin Province, 132013, China

    Cheng Wang

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  1. Yawei Xu
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  2. Yan Zhou
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Correspondence to Yawei Xu.

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Editor: Tetsuji Okamoto

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Xu, Y., Zhou, Y., Yin, R. et al. A novel 76-mer peptide mimic with the synergism of superoxide dismutase and glutathione peroxidase. In Vitro Cell.Dev.Biol.-Animal 54, 335–345 (2018). https://doi.org/10.1007/s11626-018-0240-z

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