Antioxidant activity of Bios-p peptide analogue in HEK293T cells and three-dimensional structure prediction


Studies had indicate that excessive production of reactive oxygen species (ROS) affect cellular signaling pathways, which is associated with pathological and physiological conditions such as cancer, diabetes and neurodegenerative diseases In this context, our laboratory has obtained the Bios-p, a ROS modulator, peptide analogue by sequencing from the seed of Bauhinia bauhinoides, which represents the active 12-amino acid, obtained from the inhibitor BbKI protease and we predicted the three-dimensional structure of Bios-p analogue peptide using homology modeling, being patented by the working group of Dr. Maria Luiza Vilela Oliva of UNIFESP, Brazil (a member of our cluster). The protective effect on the viability and antioxidant capacity of Bios-p was studied in HEK 293T cells under oxidative stress induced by hydrogen peroxide (H2O2) using SYTOXGREEN/DHE and luminescence assay. The three-dimensional structure of Bios-p peptide analogue was predicted by homology-based modeling using Modeller9v8. The pretreatment with different concentrations of Bios-p (1 μM - 10 μM) showed an increase of 53.83% ± 3.86% the cellular viability in under oxidative stress compared to control. Furthermore, the results to indicate that HEK293T cells by incubating for 24 h with Bios-p shown a significant decreased of basal extracellular ROS on total cell population in 89.67% ± 0.76%, compared to control in the absence of the analogue. Similarly it is observed that Bios-p has a significant antioxidant effect on extracellular ROS production when cells are subjected to oxidative stress induced by 200 μM H2O2 in 64.37% ± 4.63%, compared to control in absence of H2O2 and Bios-p. These results suggest that Bios-p has potential as antioxidant agent in cells HEK293T under H2O2-induced oxidative stress and that can protect the cells viability as concentration-dependent, and we propose a new biotechnological tool for modulate the ROS production.

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Manzano, P. , Fernández, I. , Zambrano, F. , Gómez, P. , Troncoso, M. , Marty, X. , Barrera, A. and Mejía, F. (2013) Antioxidant activity of Bios-p peptide analogue in HEK293T cells and three-dimensional structure prediction. Advances in Bioscience and Biotechnology, 4, 55-61. doi: 10.4236/abb.2013.41008.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Ajila, C.M. and Prasada Rao U.J. (2008) Protection against hydrogen peroxide induced oxidative damage in rat erythrocytes by Mangifera indica L. peel extract. Food and Chemical Toxicology, 46, 303-309. doi:10.1016/j.fct.2007.08.024
[2] Ames, B.N., M.K. Shigenaga, and T.M. Hagen (1993) Oxidants, antioxidants, and the degenerative diseases of aging. Proceedings of the National Academy of Sciences of the United States, 90, 7915-7922. doi:10.1073/pnas.90.17.7915
[3] Korhonen, H. and A. Pihlanto (2003) Food-derived bioactive peptides—Opportunities for designing future foods. Current Pharmaceutical Design, 9, 1297-1308. doi:10.2174/1381612033454892
[4] Neves, S.A., et al. (2001) Neutrophil migration induced in vivo and in vitro by marine algal lectins. Inflammation Research, 50, 486-490. doi:10.1007/PL00000222
[5] Megias, C., et al. (2004) Purification of an ACE inhibittory peptide after hydrolysis of sunflower (Helianthus annuus L.) protein isolates. Journal of Agricultural and Food Chemistry, 52, 1928-1932. doi:10.1021/jf034707r
[6] Chen, J.Y., et al. (1998) α-Tocopherol Content and Oxidative Stability of Egg Yolk as Related to Dietary α-Tocopherol. Journal of Food Science, 63, 919-922. doi:10.1111/j.1365-2621.1998.tb17927.x
[7] Chan, K.M. and E.A. Decker (1994) Endogenous skeletal muscle antioxidants. Critical Reviews in Food Science and Nutrition, 34, 403-426. doi:10.1080/10408399409527669
[8] Vaz, A.M.S.F. and Tozzi, A.M.G.A. (2005) Sinopse de Bauhinia sect. Pauletia (Cav.) DC. (Leguminosae: Caesalpinioideae: Cercideae) no Brasil. Revista Brasileira de Botanica, 28, 477-491. doi:10.1590/S0100-84042005000300006
[9] Cagliari, C.I., et al. (2003) Action of Bauhinia bauhinioides synthetic peptides on serine proteinases. Biochemical and Biophysical Research Communications, 311, 241-245. doi:10.1016/j.bbrc.2003.09.203
[10] Se-Kwon, K., et al. (2011) Proteins and Peptides as antioxidants, in bioactive food proteins and peptides. CRC Press, Boca Raton, 97-116.
[11] Fink, B., et al. (2004) Detection of intracellular superoxide formation in endothelial cells and intact tissues using dihydroethidium and an HPLC-based assay. American Journal of Physiology—Cell Physiology, 28, C895-C902. doi:10.1152/ajpcell.00028.2004
[12] Eswar, N., et al. (2006) Comparative protein structure modeling using Modeller. Current Protocols in Bioinformatics—Current Protocols, Chapter 5, Unit 5-6.
[13] Laskowski, R.A., et al. (1993) PROCHECK: A program to check the stereochemical quality of protein structures. Journal of Applied Crystallography, 26, 283-291. doi:10.1107/S0021889892009944
[14] Melo, F., et al. (1997) ANOLEA: A www server to assess protein structures. Proceedings International Conference on Intelligent Systems for Molecular Biology, 5, 187-190.
[15] Villegas, J., et al. (2005) Influence of reactive oxygen species produced by activated leukocytes at the level of apoptosis in mature human spermatozoa. Fertility and Sterility, 83, 808-810. doi:10.1016/j.fertnstert.2004.09.022
[16] Blanchetot, C. and J. Boonstra (2008) The ROS-NOX connection in cancer and angiogenesis. Critical Reviews in Eukaryotic Gene Expression, 18, 35-45. doi:10.1615/CritRevEukarGeneExpr.v18.i1.30
[17] Hansen, D., et al. (2007) Crystal structure of a novel cysteinless plant Kunitz-type protease inhibitor. Biochemical and Biophysical Research Communications, 360, 735-740. doi:10.1016/j.bbrc.2007.06.144
[18] Vitkup, D., et al. (2001) Completeness in structural genomics. Nature Structural & Molecular Biology, 8, 559-566. doi:10.1038/88640

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