Research Articles

2018  |  Vol: 4(3)  |  Issue: 3(May-June)  |  https://doi.org/10.31024/ajpp.2018.4.3.4

Studies on Cytotoxicity and Antioxidant activities of Lectin-like proteins from Phytoparasites (Cuscuta europaea)

Komola A. Kakhorova^a*, Zaynat S. Khashimova^a, Ekaterina O. Terenteva^b

^aInstitute of Bioorganic Chemistry, Uzbek Academy of Sciences, M. Ulughbek Str. 83, 100125 Tashkent, Uzbekistan.

^bInstitute of the Chemistry of Plant Substances, Uzbek Academy of Sciences, M. Ulughbek Str. 77, 100170 Tashkent, Uzbekistan.

*Corresponding author

Komola A. Kakhorova

Institute of Bioorganic Chemistry,

Uzbek Academy of Sciences, M. Ulughbek Str. 83, 100125 Tashkent, Uzbekistan.

Abstract

Objective: Lectins are of high interest among glycoproteins, functionally vital proteins of both plant and animal cells. Lectin-like proteins (LLP) from phytoparasites are not much explored. The work was initiated to study cytotoxic and antioxidant activities of lectin-like glycoproteins isolated from the dodder (Cuscuta europaea). Material and methods: LLP were isolated from the seeds of the plant by extraction with phosphate buffer solution with subsequent gradient precipitation with 20% and 50% ammonium sulfate. After centrifugation the supernatants: S₂₀ and S₅₀, as well as LLP₂₀ and LLP₅₀ precipitations were dialyzed against the distilled water. The fractions have been established to manifest the highest hemagglutinating activity. The carbohydrate specificity was studied in these fractions to demonstrate LLP₅₀ glucose- and mannose-specificity. Cytotoxic activity of LLP was assessed biochemically in B-16 (skin cancer) and C127 (breast cancer) cell cultures by MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) method and calculation of viable cells, and morphologically. Results: Comparative analysis of cytotoxic effect produced by various LLP fractions on B-16 and C-127 cells demonstrated the most pronounced cytotoxic effect of the proteins in B-16 cells, that melanoma B-16 cells are more sensitive to Cuscuta europaea LLP influence.  S₂₀ and LLP₂₀ fractions of dodder showed little antioxidant properties reducing the level of MDA, and also activating antioxidant protective enzymes - superoxide dismutase. Conclusions: Thus, we isolated and characterized LLP form Cuscuta europaea, as well as demonstrated cytotoxic effect of LLP₂₀ and LLP₅₀ on C127 and B-16 cells. For the first time cytotoxic and antioxidant activities of LLP from dodder plant Cuscuta europaea was studied.

Keywords: Cuscuta europaea, cytotoxic activity, antioxidant activity, lectin-like glycoproteins